Clinical trial NCT03895996
Conditions: Type 1 Diabetes Mellitus
Sponsor: Avotres Inc.
Start date: 2019-06-20; Completion date: 2023-12-19
Enrollment: 25 participants; Allocation: RANDOMIZED; Intervention model: PARALLEL

Brief summary:
This is a double-blind, randomized , placebo-controlled study to evaluate the safety and tolerability of AVT001, and to assess AVT001 as a potential treatment for type 1 diabetes (T1D). The trial will involve approximately 24 new-onset T1D subjects.

Detailed description:

Arms: arm 1: Infusion of AVT001 (treatment) arm 2: Infusion of AVT001-matched placebo
Interventions: intervention 1: AVT001 intervention 2: Placebo
Intervention descriptions: intervention 1: autologous dendritic cell therapy intervention 2: matched placebo

Protocol documents:
Prot_SAP_000.pdf: 
Protocol 
 
This trial protocol has been provided by the authors to give readers additional information about 
their work. 
 
 
Protocol for: Jason L. Gaglia, Heather L. Daley, Nora K. Bryant, et al. Novel Autologous Dendritic Cell 
Therapy AVT001 for Type 1 Diabetes. NEJM Evid. DOI: 10.1056/EVIDoa2300238. 


Cover Page 
This supplement contains the following items: 
1. Original protocol, ﬁnal protocol, summary of changes.
More speciﬁcally, it contains:
•
Original Protocol for Avotres’ T1D trial (AVT001-T1D-01), ﬁnalized on Jul 15th, 2013,
•
Latest version of Protocol (Amendment 3) for Avotres’ T1D trial (AVT001-T1D-01) reported in the
paper, ﬁnalized on Feb 1st, 2021,
•
Summary of Change, Protocol Amendment 3 vs 2, 2 vs 1 and 1 vs Original.
2. Original staƟsƟcal analysis plan, ﬁnal staƟsƟcal analysis plan, summary of changes.
More speciﬁcally, it contains:
•
Original StaƟsƟcal Analysis Plan (SAP), ﬁnalized on Mar 28th, 2022,
•
Latest version of StaƟsƟcal Analysis Plan (SAP) for the Day 360 analysis of Avotres’ T1D trial
(AVT001-T1D-01) reported in the paper, ﬁnalized on Dec 2nd, 2022,
•
Summary of Change, SAP v2.0 vs SAP v1.0.


  
 
CONFIDENTIAL 
 
Clinical Protocol 
 
A Phase I/II, Double-Blind, Placebo Controlled, Parallel Design of AVOTRES Cell-
Based Therapy as a Treatment for Human Type 1 Diabetes 
 
Principal Investigator: 
Name: Daniel S. Donovan, Jr., MD, MS, CDE 
Address: 161 Fort Washington Ave. Room 2-212 
New York, NY 10032 
Phone: 212-305-0504 
Email: dsd8@columbia.edu 
Regulatory Sponsor: 
Name: Hong Jiang, MD, PhD 
Address: Columbia University Medical Center 
PH - 8 East, Ste. 101,  
622 West 168th St  
New York, NY 10032 
Phone: 212-305-9984/201-944-3352     
Fax: 212-305-6070  
Email: hj4@columbia.edu 
Study Product: 
Autologous dendritic cells loaded with Hsp60sp peptide (AVOTRES) 
Protocol Number: 
Not Yet Assigned 
IND Number: 
Not Yet Assigned 
Co-Investigators: 
Name: Leonard Chess MD   
Address: 630 W.168th Street PH 8-east Ste 101 
Phone: 212-305-9984 
Email: lc19@columbia.edu 
 
Name: Carlos R. Lopez, MD, MS 
Address: 622 W. 168th St, Ph 9 East Room 103-C 
New York, NY    10032 
Phone:   212-305-2920 
Email: cl517@columbia.edu  
 
Biostatistician: 
Name: Rajasekhar Ramakrishnan, Eng.Sc.D. 
Address: 601 West 168 Street #54 
New York NY 10032 
Phone: 212-305-7989  
Fax: 212-305-5834  
Email: rr6@columbia.edu  
Date: 
July 14, 2013 
Version: 
Version 0.7 
Amended: 
N/A 
 


 
 
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Clinical Protocol Table of Contents 
LIST OF ABBREVIATIONS ...................................................................................................... IV 
STUDY SUMMARY ................................................................................................................... 1 
1 
INTRODUCTION ............................................................................................................... 4 
1.1 
BACKGROUND ..................................................................................................................... 4 
1.2 
FAILURE OF SELF-NONSELF DISCRIMINATION BY THE IMMUNE SYSTEM RESULTS IN 
PATHOGENESIS OF AUTOTIMMUNE DISEASE SUCH AS T1D ........................................................................ 5 
1.3 
THE AUTOIMMUNE PROCESS OF TYPE 1 DIABETES ................................................................ 5 
1.4 
AUTOREACTIVE T CELLS IN T1D ........................................................................................... 6 
1.5 
IMMUNOTHERAPEUTIC APPROACHES IMMUNOTHERAPEUTIC APPROACHES TO TREAT T1D ........ 6 
1.6 
DENDRITIC CELL BASED THERAPIES FOR T1D ........................................................................ 7 
1.7 
DOSING RATIONALE ............................................................................................................. 7 
2 
STUDY OBJECTIVES....................................................................................................... 9 
2.1 
PRIMARY OBJECTIVES ........................................................................................................ 10 
2.2 
SECONDARY OBJECTIVES .................................................................................................. 10 
2.3 
GENERAL STUDY DESIGN ................................................................................................... 10 
2.4 
STUDY DURATION .............................................................................................................. 11 
2.5 
STUDY ENDPOINTS ............................................................................................................ 11 
3 
SUBJECT SELECTION AND WITHDRAWAL .................................................................12 
3.1 
INCLUSION CRITERIA .......................................................................................................... 12 
3.2 
EXCLUSION CRITERIA ......................................................................................................... 12 
3.3 
SUBJECT RECRUITMENT AND SCREENING ........................................................................... 13 
3.4 
EARLY WITHDRAWAL OF SUBJECTS .................................................................................... 13 
4 
STUDY DRUG .................................................................................................................13 
4.1 
AVOTRES VACCINE (INVESTIGATIONAL) ............................................................................ 13 
4.2 
PREPARATION AND ADMINISTRATION OF STUDY DRUG. ........................................................ 13 
4.3 
RECEIVING, STORAGE, DISPENSING AND RETURN ............................................................... 14 
4.4 
RETURN OR DESTRUCTION OF STUDY PRODUCT ................................................................. 14 
5 
STUDY PROCEDURES ...................................................................................................14 
5.1 
INFORMED CONSENT.......................................................................................................... 14 
5.2 
SCREENING ....................................................................................................................... 15 
5.3 
LEUKAPHERESIS ................................................................................................................ 15 
5.4 
IMMUNOASSESSMENT “POTENCY” ASSAY ............................................................................. 15 
5.5 
TREATMENT REGIMEN ........................................................................................................ 15 
5.6 
BASELINE CLINICAL ASSESSMENTS ..................................................................................... 16 
5.7 
AVOTRES [STUDY PRODUCT] VACCINATIONS ................................................................. 16 
5.8 
INTERIM EVALUATIONS AND VACCINATIONS ......................................................................... 16 
5.9 
STUDY COMPLETION AND EVALUATION ............................................................................... 16 
5.10 
SUBJECT COMPLIANCE MONITORING .................................................................................. 16 
5.11 
PRIOR AND CONCOMITANT THERAPY .................................................................................. 17 
5.12 
LONG-TERM FOLLOW-UP .................................................................................................... 17 
6 
STATISTICAL PLAN .......................................................................................................17 
6.1 
SAMPLE SIZE DETERMINATION AND METHODS ..................................................................... 17 
7 
RISKS AND DISCOMFORTS ..........................................................................................18 
7.1 
VENIPUNCTURE ................................................................................................................. 18 
7.2 
LEUKAPHERESIS ................................................................................................................ 18 


 
 
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7.3 
RESPONSE TO AVOTRES ................................................................................................. 18 
8 
SAFETY AND ADVERSE EVENTS .................................................................................18 
8.1 
DEFINITIONS ...................................................................................................................... 18 
8.2 
RECORDING OF ADVERSE EVENTS ...................................................................................... 20 
8.3 
REPORTING OF SERIOUS ADVERSE EVENTS ........................................................................ 21 
8.4 
REPORTING PROCESS ....................................................................................................... 22 
8.5 
OTHER REPORTABLE EVENTS ............................................................................................ 22 
8.6 
ADDITIONAL REPORTING REQUIREMENTS ............................................................................. 23 
8.7 
PROTOCOL EXCEPTIONS/DEVIATIONS ................................................................................. 24 
8.8 
STOPPING RULES .............................................................................................................. 25 
8.9 
MEDICAL MONITORING ....................................................................................................... 25 
8.10 
INTERNAL DATA AND SAFETY MONITORING COMMITTEE ....................................................... 25 
9 
DATA HANDLING AND RECORD KEEPING ..................................................................25 
9.1 
CONFIDENTIALITY .............................................................................................................. 25 
9.2 
SOURCE DOCUMENTS ........................................................................................................ 26 
9.3 
CASE REPORT FORMS ....................................................................................................... 26 
9.4 
RECORDS RETENTION ........................................................................................................ 26 
10 
STUDY MONITORING, AUDITING, AND INSPECTING ..................................................26 
10.1 
STUDY MONITORING PLAN ................................................................................................. 26 
10.2 
AUDITING AND INSPECTING ................................................................................................. 26 
11 
ETHICAL CONSIDERATIONS .........................................................................................27 
12 
STUDY FINANCES ..........................................................................................................27 
12.1 
FUNDING SOURCE ............................................................................................................. 27 
12.2 
CONFLICT OF INTEREST...................................................................................................... 27 
12.3 
SUBJECT STIPENDS OR PAYMENTS ..................................................................................... 27 
13 
PUBLICATION PLAN ......................................................................................................28 
14 
SCHEDULE OF EVENTS ................................................................................................28 
15 
APPENDICES ..................................................................................................................29 
15.1 
APPENDIX A:  MONITORING PLAN ....................................................................................... 29 
15.2 
APPENDIX B:  DATA SAFETY MONITORING BOARD CHARTER ................................................ 29 
15.3 
APPENDIX C:  COMBINATION INFORMED CONSENT AND HIPPA FORM .................................. 29 
16 
REFERENCES .................................................................................................................30 
 
 


 
 
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List of Abbreviations 
 
A1c 
Hemoglobin A1c 
APC 
Antigen Presenting Cells 
ATG 
Anti-thymocyte Globulin 
CD4, CD8 
Cluster of Differentiation  
CFR 
Code of Federal Regulations 
CRF  
Case Report Form 
CTCAE 
Common Terminology Criteria for Adverse Events 
CTL 
Cytotoxic T Cell 
DC 
Dendritic Cell 
DFCI-CMCF 
Dana Farber Cancer Institute, Cell Manipulation Core Facility 
DMK 
Dystrophia Myotonica Kinase 
DSMB 
Data Safety Monitoring Board 
GAD 
Glutamic Acid Decarboxylase 
GCP 
Good Clinical Practice 
GFAP 
Glial Fibrillary Acidic Protein 
GM-CSF 
Granulocyte Macrophage-Colony Stimulating Factor 
HbA1c 
Hemoglobin A1c 
HBsAg 
Hepatitis B Virus Surface Antigen 
HBV 
Hepatitis B Virus  
HCV 
Hepatitis C Virus 
HIV 
Human Immunodeficiency Virus 
HLA 
Human Leukocyte Antigen  
Hsp60p 
Heat Shock Protein 60 Signal Peptide 
HTLV-1/2 
Human T-lymphotropic virus Type I/2 
ICA 
Islet Cell Antibody 
iDC 
Immature Dendritic Cell 
IFNγ  
Interferon γ 
IGRP 
Islet-Specific Glucose-6-Phosphate Catalytic Subunit Related Protein 
IL 
Interleukin 
IV 
Intravenous 
mcg 
Micrograms 
MHC 
Major Histocompatability Complex  
MMTT 
Mixed Meal Tolerance Test 
MRI 
Magnetic Resonance Image 
mRNA 
Messanger Ribonucleic Acid 
NOD 
Non-Obese Diabetic (murine model) 
ppIAPP 
Pre-Pre-Islet Amyloid Polypeptide Protein 
Qa-1 
Murine MHC class I molecule 
RNA 
ribonucleic acid 
s.c. 
subcutaneous 
T1D 
Type 1 Diabetes 
TNFα  
Tumour Necrosis Factor α 
Treg cells  
Regulatory T Cells 
 


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CONFIDENTIAL 
 
Study Summary 
Title 
A Phase I/II Double-Blind, Placebo Controlled, Parallel Design of AVOTRES 
Cell-Based Therapy as a Treatment for Human Type 1 Diabetes 
Short Title 
AVOTRES: Autologous Dendritic Cells Loaded with hHsp60sp Peptide 
Protocol Number 
Not yet assigned 
Clinical Phase 
Phase I/II 
IND Sponsor 
Hong Jiang, MD, PhD 
Columbia University Medical Center 
PH - 8 East, Ste. 101,  
622 West 168th St  
New York, NY 10032 
Phone: 212-305-9984/201-944-3352     
Fax: 212-305-6070  
Email: hj4@columbia.edu 
Principal Investigator 
Daniel S. Donovan, Jr., MD, MS, CDE 
Columbia University Medical Center 
161 Fort Washington Ave. Room 2-212 
New York, NY 10032 
Phone: 212-305-0504 
Email: dsd8@columbia.edu 
Study Duration 
Treatment phase is 90 days; overall protocol period is 4 years with LTFU. 
Study Center(s) 
Columbia University Medical Center  
Objectives 
Primary Objectives are to determine the safety and tolerability of the 
AVOTRES reagent; and, to determine whether, among T1D patients with in 
vitro-reversible defects in the CD8+ T cell regulatory system, infusion of 
AVOTRES will lead to an in vivo reconstitution of this system.  
 
Secondary Objective is to determine whether, among T1D patients with in 
vitro-reversible defects in the CD8+ T cell regulatory system, infusion of 
AVOTRES will result in amelioration of  cell destruction. 
Accrual Objective 
Twenty-one (21) 
Study Product, Dose, 
Route, Regimen 
This is a double-blind, placebo controlled, parallel design of “AVOTRES” cell-
based therapy as a treatment for human type I diabetes (T1D). The trial will 
involve 21 new onset T1D patients who have been identified (a), as having a 
defect in HLA-E-restricted CD8+ T cell function associated with  cell 
destruction and (b), as showing evidence that this defect of HLA-E-restricted 
CD8+ T cells can be corrected by in vitro retriggering with AVOTRES. The 
subjects will be randomly divided into two groups with 14 subjects in the 
treatment arm to receive AVOTRES, and 7 subjects in the placebo arm to 
receive with saline. 


 
 
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Inclusion Criteria 
Patients who meet all of the following criteria are eligible for enrollment as 
study participants: 
– Diagnosis of Type 1 Diabetes by one or more of the following types of 
antibodies: anti-GAD65 (glutamic acid decarboxylase), anti–islet-cell 
antibody 512 (ICA512), anti-IA2 and anti-insulin antibody. 
– Age 18 or older and able to provide informed consent for 
participation. 
– Must have stimulated C-peptide levels 0.2 pmol/ml measured 
during a mixed meal tolerance test (MMTT) 
– Recent onset (diagnosis within the past 12 months) T1D patients who 
screen positive for a CD8+ T cell defect that is correctable in vitro by 
co-culture with immature dendritic cells loaded with self-peptide 
Hsp60sp. 
– All subjects will be using insulin at the time of enrollment. 
– If a participant is female with reproductive potential, she must be 
willing to avoid pregnancy and have a negative pregnancy test during 
the 12 months of treatment and for an additional 3 months.  
– Signed and dated written informed consent 
Exclusion Criteria 
Patients who meet any of the following criteria are not eligible for enrollment 
as study participants: 
– Uncontrolled diabetes (HbA1c >9.0%) 
– Hemoglobin <10.0 g/dL; leukocytes <3,000/uL; neutrophils <1,500/uL; 
lymphocytes <800/uL; platelets <100,000/uL 
– Current use of immunosuppressive or immunomodulatory therapies, 
including pharmacologic doses of systemic steroids 
– Coincident medical condition likely to require immunosuppressive or 
immunomodulatory therapies 
– Coincident medical condition likely to limit short term (5 year) life 
expectancy (malignancy, symptomatic coronary artery disease, recent 
stroke) 
– Prior radiation therapy, immunotherapy, or chemotherapy 
– Serologic evidence of current HIV-1 or HIV-2 infection 
– Evidence of current hepatitis B as demonstrated by HBSAg or 
circulating hepatitis B genomes 
– Serologic evidence of hepatitis C infection 
– Autoimmune disease aside from T1D 
– Pregnant or breastfeeding 
– Any chronic illness which, in the opinion of the principal investigator, 
should preclude participation in the trial 
– Adequate venous access to support leukapheresis 
– Urine Albumin Excretion < 300mg/gmCr 
– eGFR > 60 mL/min/1.73m2 


 
 
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Statistical 
Considerations 
Primary endpoint:  To determine whether, among T1D patients with in vitro-
reversible defects in the CD8+ T cell regulatory system, infusion of AVOTRES 
will lead to an in vivo reconstitution of this system.   Analysis assumes that the 
proportion of subjects given placebo exhibiting reversal at 12 months will be 
0 and that at least 60% of the subjects  receiving AVOTRES will exhibit in vivo 
evidence of reversal of the defect.  With a randomization of 2:1 subjects to 
Avotres or Placebo. N = 14 subjects randomized to  Avotres and  N = 7 
subjects to placebo. There would be 94% power to detect this difference in 
proportions of 0.6 with a one sided α <0.05 using the Fisher’s exact test.   
Allowing  for  up to 10% of subjects  being unavailable for evaluation  the total 
sample size is  increased  to N = 24.  
 
Secondary endpoint:  The secondary outcome of each participant is the area 
under the stimulated C-peptide curve (AUC) over the first 2 hours of a mixed 
meal glucose tolerance test conducted at the one-year visit. The AUC is 
computed using the trapezoidal rule that is a weighted sum of the C-peptide 
values over the 120 minutes. The weighted mean C-peptide is simply 
AUC/120 (pmol/mL). Let represent the C-peptide value for study subjects 
receiving AVOTRES and those receiving placebo, respectively. The secondary 
analysis will be conducted on the transformed C-peptide using the log 
function: log(C-peptide +1). This provides better normal distributional 
behavior of the random variable. The comparison will be based on a t-test of 
treatment effect in an ANCOVA model adjusting for gender, baseline age, and 
baseline log (C-peptide +1). There will 90% power to detect a threefold 
increase in stimulated  C-peptide with a one sided α <0.05 
 
A log rank test of the difference in the C-peptide failure hazard function 
between groups (C-peptide failure is defined as the first occurrence at which 
the 2 hour peak C-peptide < 0.2 pmol/ml during a MMTT). Also, a 
proportional hazards model will be used to compare treatment groups while 
adjusting for baseline level of C-peptide, gender, and baseline age.  
 
Longitudinal analysis of HbA1c, Insulin dose (units/kg) and blood glucose by 
treatment group using normal variance-covariance structure. 
 
Number and severity of adverse events. 
 
Change in autoantibodies. 
 


 
 
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1 Introduction 
This document is a protocol for a human research study. This study is to be conducted according to US 
and international standards of Good Clinical Practice (FDA Title 21 part 312 and International 
Conference on Harmonization guidelines), applicable government regulations and Institutional research 
policies and procedures.  
1.1 Background 
Type 1 diabetes (T1D) is a complex, chronic, T cell-mediated autoimmune disease in which the insulin 
producing β cells are destroyed. In T1D, there is a loss of tolerance to pancreatic β cells that results in a 
CD4 and CD8 T cell-dependent autoimmune process that culminates in complete destruction of insulin-
producing β cells, leading to insulin deficiency and dysregulated glucose metabolism. Diabetic patients 
manage their hyperglycemia by daily insulin injections, however this treatment does not cure the 
disease or prevent the possibility of the disease’s serious effects, which may include kidney failure, 
blindness, nerve damage, heart attack, stroke, and pregnancy complications (JDRF website). It is 
important then that other therapies to treat T1D, particularly target the primary event of the defect of 
the immune function that causes T1D, be identified and tested. As discussed below, immunotherapeutic 
approaches that could potentially treat and or prevent T1D have been moving into the clinic recently. 
Over the years, studies using the non-obese diabetic (NOD) mouse model have provided important 
insights into the immunology of T1D and have demonstrated the feasibility of preventing or curing T1D 
in an antigen-specific manner by specifically manipulating disease-relevant autoreactive T cell 
populations without compromising the immune system at large.  
However, conceptually speaking, selective control of unwanted immune responses could be achieved by 
induction of “antigen-specific tolerance”, provided that the specific antigens that elicit the unwanted 
immune responses have been identified. However, for decades, it has always been a tremendous 
challenge to identify the exact pathogenic peptide/s for any given human organ specific autoimmune 
diseases, or specific allo-antigen peptide/s responsible for graft rejection. In reality, no definitive 
answers have ever been obtained for any human autoimmune diseases yet. Furthermore, induction of 
“antigen specific tolerance” requires precise knowledge of the peptide associated with particular MHC 
molecules and highly polymorphic human HLA system inevitably increases the uncertainty of searching 
for such MHC/peptide complex in each patient. Thus, the necessity of identification of a precise 
MHC/peptide complex for each individual from an unknown and countless antigen specificity pool 
makes the induction of “antigen-specific tolerance” an unfeasible approach to specifically and effectively 
treat organ specific autoimmune diseases and/or control graft rejection in clinical immunology. In 
contrast, Qa-1/HLA-E restricted CD8+ T cells, by a simple and unified “class action” of self-nonself 
discrimination, control the unwanted immune responses, independent of the knowledge of the 
particular pathogenic peptides in autoimmune diseases and/or graft rejection for each patient, which 
are largely undetermined currently. (1, 2). AVOTRES, represents one such approach to manipulate 
autoreactive T-cell responses against all self antigens, including -cell antigens (1, 2), that we would like 
to evaluate in this clinical protocol.  Modulating the activity of self-reactive T cells to damper 
autoimmunity at a system level without damaging normal on-going anti-infection and anti-tumor 
immunity would be an ideal new approach to treat T1D.  


 
 
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1.2  Failure of self-nonself discrimination by the immune system results in 
pathogenesis of autotimmune disease such as T1D 
The pioneering work of Burnet and Medawar clearly indicated that self-nonself discrimination is not 
achieved by recognizing the structural differences between self antigens and foreign antigens, because 
introducing a foreign allo transplantation antigen into the immune system early enough in life renders 
the recipient animals tolerant to that foreign antigen which is subsequently treated as if it were a self 
antigen (3, 4). It is well known now that thymic negative selection that deletes high avidity T cells 
specific to any antigens, self-antigens as natural cases, is the major mechanism of the specific tolerance 
established (5-8). However, as a biological consequence of natural thymic negative selection, self-
reactive T cells with intermediate and low avidity escape into the periphery (9-11). The intermediate 
avidity self-reactive T cell pool in the periphery represent a potential danger of pathogenic 
autoimmunity (1, 12), which could be accidentally activated by a sufficiently high level of self-antigens to 
initiate organ-specific autoimmune diseases during infections or injuries (1, 11, 13-15) and must be 
effectively controlled.  We have thus postulated that self-nonself discrimination must continue in the 
periphery after the thymic negative selection to maintain self-tolerance (1, 12). In this regard, it has 
been shown that Qa-1/HLA-E restricted CD8+ T cell mediated regulatory pathway is important in 
maintaining peripheral selftolerance (16-24). We have further proposed and tested an “Avidity Model Of 
Peripheral T Cell Regulation”, in which Qa-1/HLA-E restricted CD8+ T cells, by selectively down-
regulating the intermediate avidity T cell pool, play a central role in control of organ specific auto-
immune diseases in the conventional soluble protein antigen system in both mice (19, 20, 22) and 
humans (23). The peripheral self-reactive T cell repertoire is devoid of high avidity T cells due to thymic 
negative selection. Therefore, selective down-regulation of the intermediate avidity T cell pool enables 
the selective suppression on the overall immune responses to self-antigens, mediated by the 
intermediate avidity T cells, but not on effective immune responses to foreign antigens dominated by 
the high avidity T cells. This way, the Qa-1/HLA-E restricted CD8+ T cells are able to specifically control 
organ-specific autoimmune diseases or establish donor specific transplantation tolerance without 
damaging on going effective anti-infection and anti-tumor immunity (2, 19, 20, 22, 23, 25). The selective 
down-regulation of the intermediate avidity T cell pool by the Qa-1/HLA-E restricted CD8+ T cells is 
accomplished by a specific recognition of the unique common target structure, the signal peptide of 
heat shock protein 60 (Hsp60sp) associated with the non-polymorphic MHC Class 1b molecule Qa-
1/HLA-E, preferentially expressed on the intermediate avidity T cells (19, 20, 22, 23). 
 
More relevant, we have recently shown a defect in the T cell regulatory pathway which normally 
controls autoreactive T cells that attack the body’s own tissues and organs (23).  A majority of people 
with Type 1 diabetes who were tested in the study were found to have this newly-identified 
cellular/molecular defect on HLA-E restricted CD8+ T c ells, and we were able to successfully correct the 
defect in-vitro in most of the patients tested by retriggering the CD8+ T cells with Avotres in vitro made 
of the autologous DC loaded with self-peptide hHsp60sp (23).  
 
Our preclinical human in vitro studies and murine in vivo studies (Please see details in Section 8) formed 
the scientific basis for this IND application for trying AVOTRES in vivo. 
 
1.3 The Autoimmune Process of Type 1 Diabetes 
Development of autoimmune disorders is a result of breakdown in mechanisms that maintain 
unresponsiveness to self. Specifically, T1D is an automimmune disorder in which T cells are self-reactive 
or activated T cells that are activated against major antigenic components of pancreatic β cells are out of 


 
 
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the control by the normal regulatory mechanisms. While these self-reactive T cells are under the control 
of peripheral regulatory mechanisms in healthy individuals, failure of control leads to the destruction of 
the β cells and consequent T1D. The autoimmune attack on pancreatic cells is orchestrated by a 
variety of cells that produce cytokines and other toxic mediators. These self-reactive T cells work 
together with other lymphocytes and antigen-presenting cells to mediate this damage and have been 
shown in animal models to be important both in the early stages of diabetes development and in the 
final effector stages. They kill self-reactive T cells specific to self-antigens, which include  cell antigens, 
insulin and other key proteins associated with beta cell functions.  
1.4 Autoreactive T cells in T1D  
As reviewed in Wong 2008, many studies carried out in the NOD mouse model of T1D have indicated 
that both CD4+ and CD8+ T-cells inflict islet cell damage both at an early stage in diabetes 
development and at the final effector phase (26-28). There appears to be considerable similarity 
between the antigens recognized in humans and in the NOD animal models. In human studies, so far 
limited to testing peripheral blood lymphocytes, a variety of reactivities to putative autoantigens have 
been found, including proinsulin, GAD65, ppIAPP (pre-pro-islet amyloid polypeptide protein) and GFAP 
(glial fibrillary acidic protein). By the time diabetes occurs, it appears that there may be a broad spread 
of reactivity, which is different for each patient, and it will continue to be a major challenge to identify 
those antigenic epitopes that are important in the early phases of pathogenesis. Many of these antigenic 
epitopes represents attractive targets to go after for T1D therapeutic interventions.  
 
Mechanisms by which T cells cause damage include production of cytotoxic cytokines such as IFNγ 
(interferon γ) and TNFα (tumour necrosis factor α), that have a direct cytotoxic action on the islets. In 
addition, these cytokines have an important role in up-regulation of MHC class I molecules, which could 
increase immune recognition of the cells. These T-cell-generated inflammatory cytokines can work 
together with cytokines produced by macrophages, such as IL (interleukin)-1, to have a synergistic 
damaging effect on islet -cells. The CD8+ CTLs also exocytose granules from secretory lysosomes which 
contain cytotoxic proteins that include the pore-forming protein, perforin, and serine esterases, called 
granzymes. In summary, autoreactive T cells are considered as emerging targets for prospective antigen-
specific therapeutic interventions. 
1.5 Immunotherapeutic approaches Immunotherapeutic approaches to treat T1D  
Many broad based immunosuppressive and antigen-specific immunoregulatory therapies have been and 
are currently being evaluated for their utility in the prevention and treatment of T1D. These may be 
administered as monotherapeis or in combination. Some of the below-mentioned have shown efficacy 
in initial clinical trials. Many tend to be small molecule inhibitors and monoclonal antibodies directed 
against cytokine and cytokine receptors, proteases, CD3 or CD20 receptors (i.e. Rituximab), GAD65, 
CTL14, heat shock proteins, or insulin-related molecules (29). Systemic non-antigen specific 
immunomodulator interventions include cycyclosporine A, azatiopirine, and anti-thymocyte globulin 
(ATG) plus prednisolone. 
Other cell therapies using dendritic cells are also currently being tested as described briefly below and in 
Section 3.7. Additional examples of cell therapies include targeting regulatory T cells (Tregs). Novel 
approaches to isolate and expand polyclonal and antigen-specific Tregs in vitro have been developed for 
immunotherapy and a clinical trial to test this hypothesis is now underway in the US (Clinicaltrials.gov 
identifier NCT01210664). 


 
 
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Unlike those small molecule therapies and any currently tried cell-based therapies, Avotres provides for 
a new type of cell-based therapeutic approach to treat and/or prevent T1D. Avotres, used as a vaccine, 
activates and corrects the defective regulatory pathway mediated by HLA-E restricted CD8+ T cells in the 
identified patients that normally function to selectively control the pathogenic self-reactive T cells that 
destroy cells without damaging the normal on going anti-infection and anti-tumor immunity, a process 
called self-nonself discrimination (19, 20, 22, 23). 
1.6 Dendritic cell based therapies for T1D 
Dendritic cells (DCs) were first described by Ralph Steinman (30) nearly forty years ago.  is known that 
DCs existed in all lymphoid and most non-lymphoid tissues. DCs are antigen-presenting cells (APCs), 
which play a critical role in the regulation of the adaptive immune response. To perform this function, 
DCs are capable of capturing antigens, processing them, and presenting them on the cell surface along 
with appropriate costimulation molecules. The crucial function of DCs falls broadly into two categories, 
each of which involves antigen presentation. The first category of DCs function is antigen presentation 
and activation of T cells. The second category of DCs function has been suggested that a different class of 
DCs exist with the function of inducing and maintaining immune tolerance.  
 
Cell therapy using dendritic cells use “immature” dendritic cells that are in vitro derived from monocyte 
precursors isolated from diabetic subjects and then either modified with and reinjected into the same 
individuals with the hope of resetting the immune system balance in the individual. There is a great deal 
of interest in how DCs might be exploited as a form of immunotherapy. DCs are being studied as 
adjuvants for vaccines or as a direct therapy to induce immunity against cancer. Human clinical trials are 
ongoing to use DCs to induce immunity to antigens against prostate cancer, lung cancer, melanoma and 
multiple myeloma etc. The first attempt to use DCs as cancer vaccines in humans was made by Edgar 
Engleman and Ronald Levy (31), who isolated DCs from patients with non-Hodgkin's lymphoma who had 
failed conventional chemotherapy, loaded the cells with immunoglobulin idiotype obtained from the 
patient's tumor, and re-injected the antigen-loaded cells back into the patients. Four patients involved in 
this pilot clinical trial and all treatments were well tolerated, no significant side effects were associated 
with the infusion of antigen-loaded DCs. Since then, pilot clinical trials of antigen-pulsed DCs have been 
conducted in various types of cancer, including melanoma, prostate cancer, multiple myeloma, and non-
small cell lung cancer. DC-based therapy as immunotherapy is also tried in human type 1 diabetes and a 
clinical trial to test this hypothesis has finished its phase I study in the US (Clinical Trials. gov identifier 
NCT00445913) (32). In general, it is widely accepted that DC-based therapy is consider a very safe 
therapy (Please see Section 8.2.3)  
 
1.7 Dosing Rationale 
Our preclinical human in vitro studies and murine in vivo studies formed the scientific basis for this 
clinical application to assess AVOTRES in vivo.  AVOTRES is the study product name for autologous 
immature dendritic cells loaded with self-peptide Hsp60sp.  The study product represents an 
individualized preparation of autologous immature dendritic cells, derived by culture of the subject’s 
adherent primary monocytes with GM-CSF and IL-4 for 6 days, and loaded passively with the synthetic 
oligopeptide Hsp60sp (QMRPVSRVL).  In this study design described in this protocol, three vaccines will 
be administered 30 days (+/- 2 days) apart. 


 
 
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We feel strongly that the selection of the proposed route of administration, dose levels, dosing regimen, 
timing of vaccination relative to disease state, etc. in our clinical trial should be more rely on the 
information from similar human clinical trials than animal studies. Particularly, it should not be heavily 
dependent on NOD/T1D model, which is a spontaneous disease animal model involving multiple 
mechanisms other than Qa-1 restricted CD8+ T cells and may not be exactly the same as human T1D 
patients that we plan to test. For example, the patients we propose to test are clearly defined to be the 
new onset T1D patients who have the defect of the HLA-E restricted CD8+ T cells and the defect can be 
corrected in vitro by retriggered with AVOTRES. This design is based on the studies from human T1D 
patients and its clinical regimen cannot be exactly mimicked and represented by the studies from 
NOD/T1D model. The best example in this case, which is almost identical to our circumstances, would be 
the recently completed clinical trial of an iDC based therapy on T1D patients (ClinicalTrials. gov identifier 
NCT00445913) (32) versus its preclinical studies on NOD/T1D model (33), in that injections of 10 x 106 
iDC every 2 weeks for total four times in humans versus a single injection of 2-3× 106 iDC in NOD mice. 
In NOD mice we have used 0.5-2 x 106/mouse without any abnormality in protected mice, which dose 
range would be proportionally about 50-100 x 106/person in humans.  However, in search for the 
information in other similar human clinical trials, the dosage range used is between 2-50 x 106 /person 
every 2weeks for 2-13 times and showed no significant side effects (See Table below). Particularly, in 
one recently completed trial using iDC onT1D patients, 10 x 106 iDC every 2 weeks for total four times 
were used (32), which is very close to our proposal, eg, 10 x 106 iDC every 4 weeks for total three times 
will be used. Finally, from our own studies, HLA-E restricted CD8+ T cells represent a very potent subset 
of CD8+ T cells, probably account for 1-2% of total CD8+ T cells. We calculate if the iDCs needed for 
activate such CD8+ T cells is at a ratio of 1:50-100, 10 x 106 of iDC would activate 500-10,000 x 106 of 
HLA-E restricted CD8+ T cells, which is in the range of 1-2% of total 10x1012 CD8+ T cells.  
 
The details on the rationale for the dosing regimen based on Clinical Trial (NCT00445913): 
The product AVOTRES, autologous immature DC (iDC) loaded with self-peptide Hsp60sp, administered in 
our first-in-human safety study is, indeed, a first in human investigational product. We agree that the 
referenced trial is not the exact product, however represents the same class of immature dendritic cell 
product, and is the most similar investigational therapy that is currently in clinical testing. We therefore, 
are using this particular Clinical Trial (NCT00445913) as the basis for us to design the dosing regimen for 
AVOTRES for the following reasons: 
 
a). Both products are autologous iDC treated, in vitro, with either antisense oligonucleotides (AS-
ODN), or in our case self-peptide, Hsp60sp. Although, they are not exactly the same reagents and may 
work by different mechanisms, they are basically the same type of biomedical agents, which suggests 
that they are biologically and structurally very similar in their safety considerations.  
 
b). Importantly, compared with iDC treated with AS-ODN in Clinical Trial (NCT00445913), AVOTRES 
even more closely mimics the natural form of iDC in vivo, because under certain circumstances iDC can 
present endogenous Hsp60sp. It could be a natural form of AVOTRES, except the conditions of how the 
process can be controlled is unknown and need to be thoroughly studied. AVOTRES is made under the 
condition that Hsp60sp was loaded at an excessive amount to iDC to make sure any other Qa-1/HLA-E 
binding peptides cannot compete with Hsp60sp. So, conceptually speaking, Avotres may be even safer 
than the product of iDC treated with AS-ODN in Clinical Trial (NCT00445913). 
 
c). Both products were tried in preclinical studies in T1D/NOD mice model with almost identical 
dosing regimen and showed similar in vivo efficacy (Please see 8.2.2.2.2, Page172-175) (32,33). 


 
 
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d). In a dozen of DC-based therapies in anti-tumor clinical trials, the commonly used products are 
autologous DC loaded with different tumor antigen peptides with the dosing regimen in the range 
between 2-50 x 106 /person every 2 to 4 weeks for 2-13 times and showed no significant side effects, 
compared with which our dosing regimen of AVOTRES falls within the range (Please see 8.3.2, Page 177-
178)  
 
Table 1.7.1. Summary of dosage and route of administration of dendritic cell-base vaccines in completed 
human clinical trails, which have been concluded as safe therapies. 
 
Therapy to 
Dose/schedule  
Route 
No. 
Patients 
Ref. 
NHL 
(Non-Hodgkin’s 
lymphoma) 
2-17×106 / 4 weeks × 4 
i.v. 
35 
(34) 
 
2-17×106 / 4 weeks × 4 
i.v. 
10 
(31) 
Melanoma 
5-50×106 / 2 weeks × 4 
i.v./i.d./i.n. 
28 
(35) 
 
5-50×106 / 2 weeks × 4 
i.v. 
14 
(36) 
 
3×106+6/12×106  / 2 weeks × 5 
i.d.+i.v. 
13 
(37) 
Myeloma 
0.5-11×106 / 4 weeks × 2 
i.v. 
12 
(38) 
 
3.5-89×106 / 2 weeks × 3 
i.v. 
6 
(39) 
Prostate cancer 
1-20×106 / 6-8 weeks × 4-5 
i.v. 
51 
(40) 
 
10×106/ 2 weeks × 4 
i.v.+i.d. 
8 
(41) 
 
0.3-40×106 / 4 weeks × 2 
i.v./i.d./i.l. 
21 
(42) 
 
10×106/ 2 weeks × 3 
i.v.+i.d. 
13 
(43) 
 
0.94-2.02×108/ 4 weeks × 3 
i.v. 
14 
(44) 
RCC 
(Renal cell 
carcinoma) 
5-10×106 / 4 weeks × 3-13 
i.v./i.d. 
35 
(45) 
 
10-50×106 +10×106  /2 weeks × 
3 
i.v.+i.d. 
10 
(46) 
Type 1 Diabetes 
10×106 / 2 weeks × 4 
i.d. 
10 
(32) 
 
Based on the information from our own studies and the published results from clinical trials on DC-
based therapies we have chosen a relatively low dose and regimen of AVOTRES for safety consideration 
with a possible maximum efficacy in vivo in our proposal.  
We choose to vaccinate newly diagnosed T1D patients that will be determined to have the defect of 
HLA-E restricted CD8+ T cells and the defect could be corrected by co-culture with AVOTRES in vitro.  
 
2 Study Objectives 
Our working model for maintenance of T cell self-tolerance holds that a subset of HLA-E restricted 
regulatory CD8+ T cells that specifically recognize the target structure of Hsp60sp associated with HLA-E 
expressed on the pathogenic self-reactive T cells and selectively down-regulate these pathogenic T cells 


 
 
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which are responsible for autoimmune diseases (19, 20, 22, 23). The failure of this HLA-E-restricted 
CD8+ T cell subset to recognize, or to suppress, its target population results in a permissive state in 
which organ-specific autoimmunity can emerge. We have identified defects in this CD8+ T cell system 
among nine of ten T1D subjects evaluated, and found the defect to be reversible in most of these in 
vitro by co-incubation with Hsp60sp-loaded autologous immature dendritic cells (AVOTRES).  
2.1 Primary Objectives 
– To determine the safety and to Tolerability of the AVOTRES reagent. 
– To determine whether, among T1D patients with in vitro-reversible defects in the CD8+ T 
cell regulatory system, infusion of AVOTRES will lead to an in vivo reconstitution of this 
system.  
2.2 Secondary Objectives 
– To determine whether, among T1D patients with in vitro-reversible defects in the CD8+ T 
cell regulatory system, infusion of AVOTRES will result in amelioration of  cell destruction. 
 
The justification for the study design derives from the primary objective of the proposed trial to assess 
the safety and efficacy of this first-in-man therapy. The design exposes only small numbers of patients 
separated by defined intervals. The built-in delays in enrollment avoid exposing multiple individuals 
simultaneously as potential toxicities are assessed in a step-by-step fashion. The cohort size is chosen to 
allow detection of high frequency severe adverse events at the dose given and also the possible efficacy 
by this vaccine.  
2.3 General Study Design 
Participants will be randomized to receive AVOTRES or placebo (saline) by intravenous infusion. 
Administration of AVOTRES will be performed in a double-blinded fashion as follows: the dosage 
regimen will be 1 x 107 cells/infusion for three infusions 30 (+/- 2) days apart. Each infusion will be 
administered by qualified clinical personnel in an established clinical research facility. Patients will be 
monitored for physical complaints as well as for heart rate, blood pressure, and arterial oxygen 
saturation and changes in physical exam throughout the infusion procedure and for a minimum of 2 
hours following the procedure. Although unlikely to be required, the clinical facility will include at the 
patient’s side all medications and equipment necessary to treat acute anaphylactoid/infusion reactions, 
respiratory distress, and cardiac arrhythmia.  
 
The saline placebo group was selected for following reasons: 
 
(a) AVOTRES was well characterized in our preclinical human in vitro studies and three animal models 
for in vivo studies, where several control groups for AVOTRES were tested, including autologous DC 
loaded with B7 (human) or Qdm (mouse), and DC from Qa-1 KO mice loaded with Hsp60sp or Qdm 
(Please see Section 8 for details). The animal studies were performed in inbred mice of sufficiently 
large size of the experiments with statistically significant differences. Thus, to repeat such types of 
controls in human clinical trials is unnecessary and extremely costly and would not provide any 
more useful information. 
(b) It is well know that DC is the most potent antigen presenting cells to evoke a verity of immune 
responses under different circumstances, including presenting self-peptide/s generated 
endogenously by the DC itself. For example, it can present endogenous Hsp60sp in certain 
conditions such as under stress. It could be a natural form of AVOTRES, except the conditions of how 


 
 
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the process can be controlled is unknown and need to be thoroughly studied.  AVOTRES is made 
under the condition that Hsp60sp was loaded at an excessive amount to make sure any other Qa-
1/HLA-E biding peptide cannot compete with Hsp60sp. Thus, only DC vaccine cannot be considered 
as a proper control for AVOTRES as placebo group either. 
(c) For above reasons, untreated subjects with only infusion of saline would be an ideal control as 
placebo group that can provide clean background of evaluation of efficacy for AVOTRES treatment. 
 
 
2.4 Study Duration 
The AVOTRES study product administration will be completed on three separate visits within 60 days of 
the screen visit. Following completion of the three infusion procedures, all study participants will return 
at 60-90 day intervals for 12 months then every 6 months through month 24 (see ClinicalTrials.gov 
identifier NCT00445913) (32)  for repeat evaluation of the HLA-E-restricted CD8+ T cell regulatory 
activity (“potency assay”) as described in Section 5.4. 
 
All study participants will have a repeat clinical evaluation of diabetes at 1 month, 3 months, 6 months, 
12 months, 18 months and 24 months, including MMTT stimulated C-peptide, glucose tolerance, 
glycosylated hemoglobin, and autoantibody levels. 
 
All patients will be contacted by the Principal Investigator or authorized representative via clinic visit or 
telephone every 6 months after the last vaccine for up to four years to determine survival status and 
disease status.  
 
2.5 Study Endpoints 
 
Primary Study Endpoints 
(a) Safety and tolerability of the AVOTRES reagent. The primary analysis will describe the incidence 
of adverse events and   laboratory abnormalities.  
(b) Tolerability of the AVOTRES reagent. 
(c) Activity of the HLA-E-restricted CD8+ T cell regulatory function over a 1-year period following 
treatment with AVOTRES.  
 
Secondary Study Endpoints 
(a) The change in the area under the curve (AUC) of the stimulated C-peptide  levels  over the first 2 
hours of a mixed meal tolerance test (MMTT) 3 months,  6 months,  12 months, 18 months, and 
24 months  
(b) Change in A1c 
(c) Change in total daily insulin dose 
(d) Number of major hypoglycemic events (defined as loss of consciousness, seizure, or requiring 
assistance from another person because of altered state of consciousness)  
(e) Reported hypoglycemic events confirmed with capillary blood glucose measurement less than 
70 mg/dl.   
(f) Autoantibody levels   
 


 
 
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3 Subject Selection and Withdrawal 
3.1 Inclusion Criteria 
Patients who meet all of the following criteria are eligible for enrollment as study participants: 
(a) 
Diagnosis of Type 1 Diabetes by one or more of the following types of antibodies: anti-GAD65 
(glutamic acid decarboxylase), anti–islet-cell antibody 512 (ICA512), anti-IA2 and anti-insulin 
antibody. 
(b) 
Age 18 or older and able to provide informed consent for participation. 
(c) 
Must have stimulated C-peptide levels 0.2 pmol/ml measured during a mixed meal tolerance 
test (MMTT) 
(d) 
Recent onset (diagnosis within the past 12 months) T1D patients who screen positive for a 
CD8+ T cell defect that is correctable in vitro by co-culture with immature dendritic cells loaded 
with self-peptide Hsp60sp. 
(e) 
All subjects will be using insulin at the time of enrollment. 
(f) 
If a participant is female with reproductive potential, she must be willing to avoid pregnancy 
and have a negative pregnancy test during the 12 months of treatment and for an additional 3 
months.  
(g) 
Signed and dated written informed consent 
 
3.2 Exclusion Criteria 
Patients who meet any of the following criteria are not eligible for enrollment as study participants: 
(a) 
Uncontrolled diabetes (HbA1c >9.0%) 
(b) 
 Hemoglobin <10.0 g/dL; leukocytes <3,000/uL; neutrophils <1,500/uL; lymphocytes <800/uL; 
platelets <100,000/uL 
(c) 
Urine Albumin Excretion < 300mg/gmCr 
(d) 
eGFR > 60 mL/min/1.73m2 
(e) 
ALT/AST < 3x normal 
(f) 
Current use of immunosuppressive or immunomodulatory therapies, including pharmacologic 
doses of systemic steroids 
(g) 
Coincident medical condition likely to require immunosuppressive or immunomodulatory 
therapies 
(h) 
Coincident medical condition likely to limit short term (5 year) life expectancy (malignancy, 
symptomatic coronary artery disease, recent stroke) 
(i) 
Prior radiation therapy, immunotherapy, or chemotherapy 
(j) 
Serologic evidence of current HIV-1 or HIV-2 infection 
(k) 
Evidence of current hepatitis B as demonstrated by HBSAg or circulating hepatitis B genomes 
(l) 
Serologic evidence of hepatitis C infection 
(m) 
Autoimmune disease aside from T1D 
(n) 
Pregnant or breastfeeding 
(o) 
Any chronic illness which, in the opinion of the principal investigator, should preclude 
participation in the trial 
(p) 
Adequate venous access to support leukapheresis 


 
 
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3.3 Subject Recruitment and Screening 
Subjects will be recruited for this study from the clinical practices of the Columbia University Medical 
Center and its referral sources.  Subjects will be required to give written informed consent to participate 
in the study before any screening tests or evaluations are conducted.  
3.4 Early Withdrawal of Subjects 
Criteria for Premature Termination from the Study 
Subjects may withdraw from the study prior to the expected completion date for the following reasons: 
 Unacceptable toxicity and other safety reasons 
 Progression or disease relapse 
 Subject consent withdrawal 
 Decision by the investigators that withdrawing is in the patient’s best interest 
 Death 
 
Standard supportive therapy will be maintained for subjects withdrawn from active treatment.  
 
Data Collection and Follow-up of Withdrawn Patients 
Every effort will be made to collect toxicity information on withdrawn patients.  Patients withdrawn 
prematurely from the trial will be followed-up for survival and disease status by the principal 
investigator or clinical investigators via clinic visit, telephone contact, or medical records review every 6 
months for 2 years.  Other methods that will be used for follow-up will include contacting next of kin, 
contacting referring or primary physicians, use of certified letters to the patient, and social security 
index. 
4 Study Drug  
4.1 AVOTRES Vaccine (Investigational) 
Drug name and active ingredients:  The study drug represents an individualized preparation of 
autologous immature dendritic cells, derived by culture of the subject’s adherent primary monocytes 
with GM-CSF and IL-4 for 6 days, and loaded passively with the synthetic oligopeptide Hsp60sp 
(QMRPVSRVL).  
 
 
Formulation of dosage: AVOTRES will be cryopreserved in infusible cryomedia in KryoSure® 20 mL bags. 
Each infusion bag will contain 1 x 107 cells, and each patient will have 3 such bags manufactured for 
infusion. The cryopreserved cells will be manufactured at the Dana Farber Cancer Institute (DFCI) Cell 
Manipulation Core Facility (CMCF; DFCI-CMCF) and will be shipped to Columbia for infusion.  
 
Route of administration: The infusions will be administered intravenously.  
 
Frequency of administration: Three (3) infusions will be administered 30 (+/- 2) days apart. 
 
4.2 Preparation and Administration of Study Drug. 
One dose of AVOTRES will be packed into a 20ml freezing bag (KryoSure® Cryopreservation Bags) and 
store in liquid nitrogen or -120 to -140 º C freezer. The bag has the port to be connected to the infusion 


 
 
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system, 20 minutes before administration, the bag will be thawed at room temperature and directly 
connected to the infusion tube that contains saline to the patient. The speed of infusion should be slow, 
<1mL/minute. 
4.3 Receiving, Storage, Dispensing and Return 
 
Receipt of Study Product:  Study Drug will be shipped to the following address: 
 
Stem Cell Processing Lab 
NewYork-Presbyterian Healthcare Medical Centers 
622 W 168th Street 
Harkness Pavilion, HP4-418, 
New York, NY 10032 
 
Upon receipt of the of the study treatment supplies, an inventory will be performed and a drug receipt 
log filled out and signed by the person accepting the shipment.  It is important that the designated study 
staff counts and verifies that the shipment contains all the items noted in the shipment inventory.  Any 
damaged or unusable study drug in a given shipment (active drug or comparator) will be documented in 
the study files.  The investigator must notify study sponsor of any damaged or unusable study 
treatments that were supplied to the investigator’s site. 
 
Storage:   The vialed study product will be kept frozen at the aforementioned Stem Cell Processing Lab 
until immediately before thaw, reconstitution, and administration. 
 
Dispensing of Study Product:  The study product will be listed under each patient’s own unique 
identifier.  The date, patient identifier, and quantity used for each study product.  Regular study product 
reconciliation will be performed to document study product consumed and study product remaining.  
This reconciliation will be logged on the study product reconciliation form, and signed and dated by the 
study team. 
4.4 Return or Destruction of Study Product 
At the completion of the study, there will be a final reconciliation of study product shipped, consumed, 
and remaining.  This reconciliation will be logged on the study product reconciliation form, signed and 
dated.  Any discrepancies noted will be investigated, resolved, and documented prior to return or 
destruction of unused study product.  Study product destroyed on site will be documented in the study 
files. 
 
5 Study Procedures 
Study procedures are listed below and can also be found in the Schedule of Events (Section 15).  No 
study procedures will be performed prior to obtaining informed consent. 
5.1 Informed Consent  
The consent process will be conducted by qualified study personnel (the Trial or Study Coordinator 
and/or Investigator or other designee). All participants (or their legally acceptable representative) must 


 
 
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read, sign and date a consent form prior to participation in the study, and/or undergoing any study-
specific procedures.  
 
The informed consent form must be updated or revised whenever important new safety information is 
available, when indicated for a protocol amendment, and/or whenever any new information becomes 
available that may affect a patient’s participation in the study.  
5.2 Screening 
The following evaluation will be performed prior to enrollment to establish patient eligibility.  All tests 
will be required within 30 days prior to treatment: 
 Medical history including prior treatment, current medicines, and drug allergies 
 Physical exam including blood pressure, pulse, temperature, weight, and retinal exam 
 Laboratory studies comprising CBC/differential, creatinine, AST, ALT, and total bilirubin; 
including creatinine clearance and albuminuria 
 Evaluation of pancreatic  cell function by established clinical tests: mixed meal stimulated C 
peptide measurement, glucose tolerance, glycosylated hemoglobin(A1c), insulin and GAD65 
auto-antibodies  
 Evaluation of HLA-E-restricted CD8+ T cells (“potency assay”), as described in Section 5.4. 
 Pregnancy test if applicable 
 
The following evaluations will be performed within 30 days prior to treatment: 
 
Viral studies comprising HIV-1, HIV-2, HTLV-1/2, and HCV serology and HBV surface antigen, 
surface antibody, and core antibody 
5.3 Leukapheresis 
Within 30 days of screening assessments, those patients qualified for enrollment will undergo 
leukapheresis at the Columbia University’s Apheresis Unit according to standard clinical procedures. The 
apheresed product will be delivered to Dana Farber Center Cancer Institute (DFCI), Cell Manipulation 
Core Facility (CMCF) for processing to produce AVOTRES.  
5.4 Immunoassessment “Potency” Assay 
15-20% (about 40 mL) of the apheresis product will be delivered to Hong Jiang’s laboratory at Columbia 
University and used for immunoassessment research assays (“potency assay”). The potency assay will be 
performed every 60-90 days after vaccination to evaluate the efficacy of AVOTRES, in improving the 
immune function, by taking a 75 mL peripheral blood sample/time point.  
5.5 Treatment Regimen 
All patients will receive the same regimen. 
 
On day 1, the patient will receive the AVOTRES infusion. 
 
The interval between injections will be 28 days for total three times. 
 
The staggering period between individuals is 4 weeks. We will monitor the  
first  9 patients to make sure that at least 6 subjects in the treatment group will be  
observed for 4 weeks for acute and subacute adverse events.  The information obtained  
will allow the clinical PI and Sponsor to determine if the time interval of 2 weeks or 4  
weeks is sufficient, we will consult with the FDA to lengthen or shorten the 4-week  
observation of patient staggering period in the clinical trial. 


 
 
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 
After each Infusion, patients will be observed for at least 2 hour including measurement of 
vital signs prior to discharge.   
 
In subsequent Infusions, patients may be discharged immediately following injection. 
Infusions will be given at the Irving Center for Clinical of the Columbia University Medical Center.   
5.6 Baseline Clinical Assessments  
Evaluation of HLA-E-restricted CD8+ T cells, as described in Section 5.4, will be performed within 30 days 
of the first vaccination, prior to infusion.  The potency assay is performed in seven (see Section 14, 
Schedule of Events) circumstances during the overall study. (1) At the time of screening of potential 
participants. Eligibility to participate is based on the demonstration of a failure of the HLA-E-restricted T 
cell regulatory population and the ability to reverse this failure by co-incubation with AVOTRES 
(hHsp60sp-loaded iDC.). (2) To verify the functional quality of the iDC manufactured by DFCI which 
constitute the AVOTRES treatment, prior to infusion. (3) To monitor the in vivo efficacy every 60-90 days 
after the treatment with AVOTRES 
 
The following baseline evaluations will be performed only if more than 2 months time has passed from 
date of screen visit: 
 Medical history and physical exam, including vital signs 
 Laboratory comprising CBC/diff, TSH, and CMV serology.   
 Immunologic study specimens 
 C-peptide responses, insulin, glucose, HbA1c levels 
5.7 AVOTRES Vaccinations 
AVOTRES will be transported at 2-8oC to the patient bedside.   Product administration training will be 
provided to each site.  The KryoSure® Cryopreservation bag has the port to be connected to the infusion 
system. Twenty minutes before administration, the bag will be thawed at room temperature and 
directly connected to the infusion tube that contains at least 200mL of saline. The speed of infusion 
should be slow, <1mL/minute for the AVOTRES mixed with saline. 
5.8 Interim Evaluations and Vaccinations 
At the time of each subsequent vaccination, patient evaluation will include: 
 Interim Medical history, including medication review 
 Limited Physical exam, including vital signs and retinal exam. 
 Laboratory studies as noted in Section 14, Schedule of Events 
 Immune studies    
 Adverse event assessment 
5.9 Study Completion and Evaluation 
The following clinical evaluation will be performed within 30 days after the last vaccination: 
 Medical history and physical exam, including vital signs and performance status 
 C-peptide responses, insulin, glucose, HbA1c levels 
 Laboratory studies as noted in Appendix A 
5.10 Subject Compliance Monitoring 
The protocol requires frequent patient visits to the clinic for treatment and monitoring during 
participation in this trial.  In addition, patients are followed-up by telephone contact between visits.   


 
 
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5.11 Prior and Concomitant Therapy 
Study participants will continue all regularly prescribed medications, including those for glycemic 
control. The introduction of immunosuppressive agents, however, will constitute grounds for 
withdrawal from the study. 
5.12 Long-term follow-up 
All patients will be contacted by the Principal Investigator or authorized representative via clinic visit or 
telephone every 6 months after the last vaccine for up to four years to determine survival status and 
disease status.  
 
6 Statistical Plan 
6.1 Sample Size Determination and Methods  
 
Primary endpoint:  To determine whether, among T1D patients with in vitro-reversible defects in the 
CD8+ T cell regulatory system, infusion of AVOTRES will lead to an in vivo reconstitution of this system.   
Analysis assumes that the proportion of subjects given placebo exhibiting reversal at 12 months will be 0 
and that at least 60% of the subjects  receiving AVOTRES will exhibit in vivo evidence of reversal of the 
defect.  With a randomization of 2:1 subjects to Avotres or Placebo. N = 14 subjects randomized to  
Avotres and  N = 7 subjects to placebo. There would be 94% power to detect this difference in 
proportions of 0.6 with a one sided α <0.05 using the Fisher’s exact test.     Allowing  for  up to 10% of 
subjects  being unavailable for evaluation  the total sample size is  increased  to N = 24.  
 
Secondary endpoint:  The secondary outcome of each participant is the area under the stimulated C-
peptide curve (AUC) over the first 2 hours of a mixed meal glucose tolerance test conducted at the one-
year visit. The AUC is computed using the trapezoidal rule that is a weighted sum of the C-peptide values 
over the 120 minutes. The weighted mean C-peptide is simply AUC/120 (pmol/mL). Let represent the C-
peptide value for study subjects receiving AVOTRES and those receiving placebo, respectively. The 
secondary analysis will be conducted on the transformed C-peptide using the log function: log(C-peptide 
+1). This provides better normal distributional behavior of the random variable. The comparison will be 
based on a t-test of treatment effect in an ANCOVA model adjusting for gender, baseline age, and 
baseline log (C-peptide +1). There will 90% power to detect a threefold increase in stimulated  C-peptide 
with a one sided α <0.05 
 
A log rank test of the difference in the C-peptide failure hazard function between groups (C-peptide 
failure is defined as the first occurrence at which the 2 hour peak C-peptide < 0.2 pmol/ml during a 
MMTT). Also, a proportional hazards model will be used to compare treatment groups while adjusting 
for baseline level of C-peptide, gender, and baseline age.  
 
Longitudinal analysis of HbA1c, Insulin dose (units/kg) and blood glucose by treatment group using 
normal variance-covariance structure. 
 
Number and severity of adverse events. 
 
Change in autoantibodies. 


 
 
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7 Risks and Discomforts 
7.1 Venipuncture  
Patients may experience slight discomfort at the site and rarely develop an ecchymosis or superficial 
thrombosis.  
7.2 Leukapheresis  
The most common adverse reactions from leukapheresis are paraesthesias, dizziness, mild chest 
tightness, or cough related to the reinfusion of autologous red blood cells treated with citrate and 
ecchymoses at the catheter sites. 
 
Potential adverse reactions related to leukapheresis include:  
 Exacerbation of symptoms from an underlying medical condition including migraine headache, 
asthma/emphysema, or cardiovascular disease.  
 Infection at catheter sites  
 Hypotension, hypertension, or bradycardia from citrate toxicity  
 Allergic reactions  
 Anemia, if red blood cells cannot be reinfused  
7.3 Response to AVOTRES  
The response to AVOTRES would be to activate HLA-E restricted CD8+ T cells in vivo, which would in turn 
selectively down-regulate any anti-self responses to control autoimmune diseases. In animal toxicology 
study conducted by Covance, the data showed no adverse effect. In addition, in general DC-based 
therapies are considered safe therapies. 
 
8 Safety and Adverse Events 
Patients will be evaluated for evidence of toxicity from 0-4 years. 
8.1 Definitions 
Unanticipated Problems Involving Risk to Subjects  
Any incident, experience, or outcome that meets all of the following criteria:  
 Unexpected in nature, severity, or frequency  (i.e. not described in study-related documents 
such as the IRB-approved protocol or consent form, the investigators brochure, etc) 
 Related or possibly related to participation in the research (i.e. possibly related means there is a 
reasonable possibility that the incident experience, or outcome may have been caused by the 
procedures involved in the research) 
 Suggests that the research places subjects or others at greater risk of harm (including physical, 
psychological, economic, or social harm). 
 
Adverse Event 


 
 
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An adverse event (AE) is any symptom, sign, illness or experience that develops or worsens in severity 
during the course of the study.  Intercurrent illnesses or injuries should be regarded as adverse events.  
Abnormal results of diagnostic procedures are considered to be adverse events if the abnormality: 
 results in study withdrawal 
 is associated with a serious adverse event 
 is associated with clinical signs or symptoms 
 leads to additional treatment or to further diagnostic tests 
 is considered by the investigator to be of clinical significance 
 
Serious Adverse Event 
Adverse events are classified as serious or non-serious.  A serious adverse event is any AE that is:  
 fatal 
 life-threatening 
 requires or prolongs hospital stay 
 results in persistent or significant disability or incapacity 
 a congenital anomaly or birth defect 
 an important medical event 
 
Important Medical Events 
Important medical events are those that may not be immediately life threatening, but are clearly of 
major clinical significance.   They may jeopardize the subject, and may require intervention to prevent 
one of the other serious outcomes noted above.  For example, drug overdose or abuse, a seizure that 
did not result in in-patient hospitalization, or intensive treatment of bronchospasm in an emergency 
department would typically be considered serious.  
 
All adverse events that do not meet any of the criteria for serious event should be regarded as non-
serious adverse events.  
 
Adverse Event Reporting Period 
The study period during which adverse events must be reported is normally defined as the period from 
the initiation of any study procedures (Day -1) to the end of the study treatment follow-up.  For this 
study, the study treatment follow-up is defined as 30 days following the last administration of study 
treatment. 
 
Preexisting Condition 
A preexisting condition is one that is present at the start of the study.  A preexisting condition should be 
recorded as an adverse event if the frequency, intensity, or the character of the condition worsens 
during the study period. 
 
General Physical Examination Findings 
At screening, any clinically significant abnormality should be recorded as a preexisting condition.  At the 
end of the study, any new clinically significant findings/abnormalities that meet the definition of an 
adverse event must also be recorded and documented as an adverse event.  
 
Post-study Adverse Event 
All unresolved adverse events should be followed by the investigator until the events are resolved, the 
subject is lost to follow-up, or the adverse event is otherwise explained.  At the last scheduled visit, the 
investigator should instruct each subject to report any subsequent event(s) that the subject, or the 


 
 
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subject’s personal physician, believes might reasonably be related to participation in this study.  The 
investigator should notify the study sponsor of any death or adverse event occurring at any time after a 
subject has discontinued or terminated study participation that may reasonably be related to this study.  
The sponsor should also be notified if the investigator should become aware of the development of 
cancer or of a congenital anomaly in a subsequently conceived offspring of a subject that has 
participated in this study.  
 
Abnormal Laboratory Values 
A clinical laboratory abnormality should be documented as an adverse event if any one of the following 
conditions is met:  
 The laboratory abnormality is not otherwise refuted by a repeat test to confirm the abnormality 
 The abnormality suggests a disease and/or organ toxicity 
 The abnormality is of a degree that requires active management; e.g. change of dose, 
discontinuation of the drug, more frequent follow-up assessments, further diagnostic 
investigation, etc. 
 
Hospitalization, Prolonged Hospitalization or Surgery 
Any adverse event that results in hospitalization or prolonged hospitalization should be documented 
and reported as a serious adverse event unless specifically instructed otherwise in this protocol.  Any 
condition responsible for surgery should be documented as an adverse event if the condition meets the 
criteria for an adverse event.  
 
Neither the condition, hospitalization, prolonged hospitalization, nor surgery are reported as an adverse 
event in the following circumstances: 
 
 Hospitalization or prolonged hospitalization for diagnostic or elective surgical procedures for a 
preexisting condition.  Surgery should not be reported as an outcome of an adverse event if the 
purpose of the surgery was elective or diagnostic and the outcome was uneventful. 
 Hospitalization or prolonged hospitalization required to allow efficacy measurement for the 
study. 
 Hospitalization or prolonged hospitalization for therapy of the target disease of the study, unless 
it is a worsening or increase in frequency of hospital admissions as judged by the clinical 
investigator. 
8.2 Recording of Adverse Events 
At each contact with the subject, the investigator must seek information on adverse events by specific 
questioning and, as appropriate, by examination.  Information on all adverse events should be recorded 
immediately in the source document, and also in the appropriate adverse event module of the case 
report form (CRF).  All clearly related signs, symptoms, and abnormal diagnostic procedures results 
should recorded in the source document, though should be grouped under one diagnosis. 
 
All adverse events occurring during the study period must be recorded.  The clinical course of each event 
should be followed until resolution, stabilization, or until it has been determined that the study 
treatment or participation is not the cause.  Serious adverse events that are still ongoing at the end of 
the study period must be followed up to determine the final outcome.  Any serious adverse event that 
occurs after the study period and is considered to be possibly related to the study treatment or study 
participation should be recorded and reported immediately. 


 
 
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8.3 Reporting of Serious Adverse Events 
Study Sponsor Notification by Investigator 
A serious adverse event must be reported to the study sponsor by telephone within 24 hours of the 
event.  A Serious Adverse Event (SAE) form must be completed by the investigator and faxed to the 
study sponsor within 24 hours.  The investigator will keep a copy of this SAE form on file at the study 
site.  Report serious adverse events by phone, facsimile or email to: 
 
Name:   
Hong Jiang, MD, PhD 
Address:  
Columbia University Medical Center 
PH - 8 East, Ste. 101,  
622 West 168th St  
New York, NY 10032 
Phone:   
212-305-9984/201-944-3352     
Fax:  
 
212-305-6070  
Email:   
hj4@columbia.edu 
 
At the time of the initial report, provide as much of the following information as possible: 
 Study identifier 
 Study Center 
 Subject number 
 A description of the event 
 Date of onset 
 Current status 
 Whether study treatment was discontinued 
 The reason why the event is classified as serious 
 Investigator assessment of the association and expectedness between the event and study 
treatment 
 
Within the following 48 hours, the investigator must provide further information on the serious adverse 
event in the form of a written narrative.  This should include a copy of the completed Serious Adverse 
Event form, and any other diagnostic information that will assist the understanding of the event.  
Significant new information on ongoing serious adverse events should be provided promptly to the 
study sponsor 
 
IRB Notification by Investigator 
This section describes the requirements for safety reporting by investigators who are faculty, affiliated 
with a research site, or otherwise responsible for safety reporting to the Columbia IRB. The Columbia 
requires expedited reporting of those events related to study participation that are unforeseen and 
indicate that participants or others are at increased risk of harm.  The Columbia IRB will not 
acknowledge safety reports or bulk adverse event submissions that do not meet the criteria outlined 
below.  The Columbia IRB requires researchers to submit reports of the following problems WITHIN 10 
WORKING DAYS FROM THE TIME THE INVESTIGATOR BECOMES AWARE OF THE EVENT: 
 
Any adverse event (regardless of whether the event is serious or non-serious, on-site or off-site) that 
occurs any time during or after the research study, which in the opinion of the principal investigator is: 


 
 
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 UNEXPECTED AND RELATED 
o Unexpected (An event is “unexpected” when its specificity and severity are not 
accurately reflected in the protocol-related documents, such as the IRB-approved 
research protocol, any applicable investigator brochure, and the current IRB-approved 
informed consent document and other relevant sources of information, such as product 
labeling and package inserts.)  
o Related to the research procedures (An event is “related to the research procedures” if 
in the opinion of the principal investigator or sponsor, the event was more likely than 
not to be caused by the research procedures.) 
8.4 Reporting Process 
Unanticipated problems posing risks to subjects or others as noted above will be reported to the  IRB 
using the form: “Unanticipated Problems Posing Risks to Subjects or Others Including Reportable 
Adverse Events” or as a written report of the event (including a description of the event with 
information regarding its fulfillment of the above criteria, follow-up/resolution and need for revision to 
consent form and/or other study documentation). 
 
Copies of each report and documentation of IRB notification and receipt will be kept in the Clinical 
Investigator’s study file. 
8.5 Other Reportable Events 
For clinical drug trials, the following events are also reportable to the IRB: 
 Any adverse experience that, even without detailed analysis, represents a serious unexpected 
adverse event that is rare in the absence of drug exposure (such as agranulocytosis, hepatic 
necrosis, Stevens-Johnson syndrome). 
 Any adverse event that would cause the sponsor to modify the investigators brochure, protocol 
or informed consent form, or would prompt other action by the IRB to assure protection of 
human subjects. 
 Information that indicates a change to the risks or potential benefits of the research, in terms of 
severity or frequency. For example:  
o An interim analysis indicates that participants have a lower rate of response to 
treatment than initially expected. 
o Safety monitoring indicates that a particular side effect is more severe, or more 
frequent than initially expected. 
o A paper is published from another study that shows that an arm of your research study 
is of no therapeutic value. 
 Change in FDA safety labeling or withdrawal from marketing of a drug, device, or biologic used 
in a research protocol. 
 Breach of confidentiality 
 Change to the protocol taken without prior IRB review to eliminate apparent immediate hazard 
to a research participant. 
 Incarceration of a participant when the research was not previously approved under Subpart C 
and the investigator believes it is in the best interest of the subject to remain on the study. 
 Complaint of a participant when the complaint indicates unexpected risks or the complaint 
cannot be resolved by the research team. 


 
 
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 Protocol violation (meaning an accidental or unintentional deviation from the IRB approved 
protocol) that in the opinion of the investigator placed one or more participants at increased 
risk, or affects the rights or welfare of subjects. 
 
FDA Notification by Sponsor 
The study sponsor is required to report certain study events in an expedited fashion to the FDA.  These 
written notifications of adverse events are referred to as IND safety reports. The following describes the 
safety reporting requirements by timeline for reporting and associated type of event: 
 
 Within 7 calendar days 
Any study event that is: 
– associated with the use of the study drug 
– unexpected,  
– fatal or life-threatening, and  
 
 Within 15 calendar days 
Any study event that is: 
– associated with the use of the study drug, 
– unexpected, and 
– serious, but not fatal or life-threatening 
-or- 
– a previous adverse event that was not initially deemed reportable but is later found to 
fit the criteria for reporting (reporting within 15 calendar days from when event was 
deemed reportable). 
Any finding from tests in laboratory animals that:  
– suggest a significant risk for human subjects including reports of mutagenicity, 
teratogenicity, or carcinogenicity. 
 
8.6 Additional reporting requirements 
Sponsors are also required to identify in IND safety reports all previous reports concerning similar 
adverse events and to analyze the significance of the current event in light of the previous reports. 
 
DSMB Notification 
Investigators will report all SAEs to the DSMB within 10 business days regardless of expectedness or 
attribution. SAE will be input into the CRF with follow up email to DSMB. 
 
All Unexpected deaths or deaths related to the study agent must be reported within 24 hours.  All other 
deaths should be reported within 30 days in the CRF.  
 
Item 
Report to DSMB 
Do Not Report to 
DSMB 
Expected, related, grade 1, 2 , 3 or 4 
X 
 
Expected, related grade 1, 2, 3 or 4.  >60 days since last subject 
treated 
 
X 
Expected, unrelated grade 1, 2 , 3 or 4 
X 
 
1, 2, 3 or 4. >690 days since last subject treated 
 
X 


 
 
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CONFIDENTIAL 
Item 
Report to DSMB 
Do Not Report to 
DSMB 
Unexpected , related grade 1, 2 , 3 or 4 
X 
 
Unexpected, related grade 1, 2, 3 or 4 >60 days since last 
subject treated 
 
X 
Unexpected , unrelated grade 1, 2 , 3 or 4 
X 
 
Unexpected, unrelated grade 1, 2, 3 or 4. >60 days since last 
subject treated 
 
X 
Expected, related death 
X 
 
Expected, unrelated death 
X 
 
Unexpected, related death 
X 
 
Unexpected, unrelated death 
X 
 
8.7 Protocol Exceptions/Deviations 
Exception:   A one time, intentional action or process that departs from the IRB and CTSRMC approved 
study protocol, intended for one occurrence. If the action disrupts the study progress, such that the 
study design or outcome (endpoints) may be compromised, or the action compromises the safety and 
welfare of study subjects, advance documented IRB and DSMB approval is required.    
 
Deviation:   A one time, unintentional action or process that departs from the IRB and DSMB approved 
study protocol, involving one incident and identified retrospectively, after the event occurred. If the 
impact on the protocol disrupts the study design, may affect the outcome (endpoints) or compromises 
the safety and welfare of the subjects, the deviation must be reported to the DSMB within 5 business 
days and the IRB within 10 business days.   
 
Any departure from the protocol that meets the following criteria should be submitted to the Sponsor: 
 
 May/can/have affects/affected subject safety.  
 Violate eligibility 
 Dose adjustment 
 Stopping criteria 
 Affect sample size (adding more subjects, decreasing number of subjects, changing the number 
of subject in a specific arm/cohort) 
Other deviations should be explained in a memo to file (MTF; such as a subject missing a visit is not an 
issue unless a critical/important treatment or procedure was missed and must have been done at that 
specific time). The Principal Investigator should give the Sponsor at least 5 days advance notice of the 
deviation. Include the following information on the Sponsor supplied deviation form:  Study identifier, 
description of deviation from protocol and necessity for deviation.  Ensure all deviations are signed by 
the Principal Investigator (or co-investigator) and submitted to the Sponsor.    
Name:   
Hong Jiang, MD, PhD 
Address:  
Columbia University Medical Center 
PH - 8 East, Ste. 101,  
622 West 168th St  
New York, NY 10032 
Phone:   
212-305-9984/201-944-3352     
Fax:  
 
212-305-6070  


 
 
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Email:  hj4@columbia.edu 
 
The Protocol Monitor will submit the deviation to the Sponsor for review and approval. The approved 
deviation will be submitted by the Investigator to the IRB, DSMB as required. 
8.8 Stopping Rules  
Patients will be clinically evaluated for evidence of toxicity from ≥ grade 3 toxicity as determined by CTC, 
which is unmanageable, unexpected and unrelated, and attributable to protocol therapy.  Termination 
of enrollment would be considered with 2 DLTs in the first 6 evaluable subjects and 3 in the first 10 
evaluable subjects. With 10 evaluable patients enrolled, there is at least 90% power for detecting any 
previously unexpected toxicity whose prevalence in this population is at least 21%.  
 
In the event of a DLT for a given patient, doses will be discontinued, the patient clinically evaluated, and 
taken off study. The study will be stopped for DLT resulting in death of a patient. 
8.9 Medical Monitoring 
It is the responsibility of the Principal Investigators to oversee the safety of the study at his site.  This 
safety monitoring will include careful assessment and appropriate reporting of adverse events as noted 
above, as well as the construction and implementation of a site data and safety-monitoring plan.  
Medical monitoring will include a regular assessment of the number and type of serious adverse events. 
8.10  Internal Data and Safety Monitoring Committee 
The Data Safety and Monitoring Board (DSMB) will monitor the data quality and adherence to safety 
rules.  Additionally, the DSMB will review all safety/toxicity data for the trial and recommend trial 
suspension or termination as needed.  Specific details of monitoring and audit frequency will be 
included in the Monitoring Plan (see Appendix A)  but will be at least every 6 months. 
9 Data Handling and Record Keeping 
9.1 Confidentiality 
Information about study subjects will be kept confidential and managed according to the requirements 
of the Health Insurance Portability and Accountability Act (HIPAA).  Those regulations require a signed 
subject authorization (see Appendix C) informing the subject of the following:  
 
 What protected health information (PHI) will be collected from subjects in this study 
 Who will have access to that information and why 
 Who will use or disclose that information 
 The rights of a research subject to revoke their authorization for use of their PHI.  
 
In the event that a subject revokes authorization to collect or use PHI, the investigator, by regulation, 
retains the ability to use all information collected prior to the revocation of subject authorization.  For 
subjects that have revoked authorization to collect or use PHI, attempts should be made to obtain 
permission to collect at least vital status (i.e. that the subject is alive) at the end of their scheduled study 
period. 


 
 
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9.2 Source Documents 
Source data is all information, original records of clinical findings, observations, or other activities in a 
clinical trial necessary for the reconstruction and evaluation of the trial.  Source data are contained in 
source documents   Examples of these original documents, and data records include: hospital records, 
clinical and office charts, laboratory notes, memoranda, subjects’ diaries or evaluation checklists, 
pharmacy dispensing records, recorded data from automated instruments, copies or transcriptions 
certified after verification as being accurate and complete, microfiches, photographic negatives, 
microfilm or magnetic media, x-rays, subject files, and records kept at the pharmacy, at the laboratories, 
and at medico-technical departments involved in the clinical trial. 
9.3 Case Report Forms 
The study case report form (CRF) is the primary data collection instrument for the study.  All data 
requested on the CRF must be recorded.  All missing data must be explained.  If a space on the CRF is left 
blank because the procedure was not done or the question was not asked, write “N/D”.  If the item is 
not applicable to the individual case, write “N/A”.  All entries should be printed legibly in black ink.  If 
any entry error has been made, to correct such an error, draw a single straight line through the incorrect 
entry and enter the correct data above it.  All such changes must be initialed and dated.  DO NOT ERASE 
OR WHITE OUT ERRORS.  For clarification of illegible or uncertain entries, print the clarification above 
the item, then initial and date it. 
9.4 Records Retention 
It is the investigator’s responsibility to retain study essential documents for at least 2 years after the last 
approval of a marketing application in their country and until there are no pending or contemplated 
marketing applications in their country or at least 2 years have elapsed since the formal discontinuation 
of clinical development of the investigational product.  These documents should be retained for a longer 
period if required by an agreement with the sponsor.  In such an instance, it is the responsibility of the 
sponsor to inform the investigator/institution as to when these documents no longer need to be 
retained.  
 
10 Study Monitoring, Auditing, and Inspecting 
10.1 Study Monitoring Plan 
This study will be monitored according to the monitoring plan in Appendix A.  The investigator will 
allocate adequate time for such monitoring activities.  The Investigator will also ensure that the monitor 
or other compliance or quality assurance reviewer is given access to all the above noted study-related 
documents and study related facilities (e.g. pharmacy, diagnostic laboratory, etc.), and has adequate 
space to conduct the monitoring visit. 
10.2 Auditing and Inspecting 
The investigator will permit study-related monitoring, audits, and inspections by the EC/IRB, the 
sponsor, government regulatory bodies, and University compliance and quality assurance groups of all 
study related documents (e.g. source documents, regulatory documents, data collection instruments, 
study data etc.).  The investigator will ensure the capability for inspections of applicable study-related 
facilities (e.g. pharmacy, diagnostic laboratory, etc.). 
 


 
 
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Participation as an investigator in this study implies acceptance of potential inspection by government 
regulatory authorities and applicable University compliance and quality assurance offices. 
11 Ethical Considerations 
This study is to be conducted according to US and international standards of Good Clinical Practice (FDA 
Title 21 part 312 and International Conference on Harmonization guidelines), applicable government 
regulations and Institutional research policies and procedures. 
 
This protocol and any amendments will be submitted to a properly constituted independent Ethics 
Committee (EC) or Institutional Review Board (IRB), in agreement with local legal prescriptions, for 
formal approval of the study conduct.  The decision of the EC/IRB concerning the conduct of the study 
will be made in writing to the investigator and a copy of this decision will be provided to the sponsor 
before commencement of this study.  The investigator should provide a list of EC/IRB members and their 
affiliate to the sponsor. 
 
All subjects for this study will be provided a consent form describing this study and providing sufficient 
information for subjects to make an informed decision about their participation in this study.  See 
Appendix 4 for a copy of the Subject Informed Consent Form.  This consent form will be submitted with 
the protocol for review and approval by the EC/IRB for the study.  The formal consent of a subject, using 
the EC/IRB-approved consent form, must be obtained before that subject is submitted to any study 
procedure.  This consent form must be signed by the subject or legally acceptable surrogate, and the 
investigator-designated research professional obtaining the consent.  
 
12 Study Finances 
12.1  Funding Source 
This trial is funded by China Institute of Strategy and Management Lanmeng Investment Co.,   
Ltd (CISM Lanmeng Investment Co., Ltd) ” in Beijing, China. 
12.2  Conflict of Interest 
The regulatory sponsor of this study (Dr. Hong Jiang), who is the person who reports to the Food and Drug 
Administration and to the University about the status and results of the study, invented the AVOTRES 
technology. She has not received money for her invention related to this technology; however, there is a 
potential to do so in the future.  The AVOTRES technology has been licensed to a for-profit company. As a 
result, these investigators have benefited and it is anticipated that they will continue to benefit financially.  
In addition, the Columbia University has a financial interest in AVOTRES. As a result of the relationship with a 
commercial entity, Columbia University is expected to benefit financially in the future. 
12.3 Subject Stipends or Payments 
Patients will not be paid for participating in this clinical trial.  However, assistance may be provided with 
parking and transportation and the costs of laboratory tests. 
 


 
 
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13 Publication Plan 
Neither the complete nor any part of the results of the study carried out under this protocol, nor any of 
the information provided by the sponsor for the purposes of performing the study, will be published or 
passed on to any third party without the consent of the study sponsor.  Any investigator involved with 
this study is obligated to provide the sponsor with complete test results and all data derived from the 
study. 
 
 
14 Schedule of Events 
 
VISIT 
NAME 
SCREEN APH 
VISIT  
1 
VISIT 
2 
VISIT 
3 
VISIT  
4 
VISIT  
5 
VISIT  
6 
VISIT  
8 
VISIT 
9 
VISIT 
10 
PROCEDURES 
SCREEN APH VAC#1 VAC#2 VAC#3 
F/U 
1 
F/U 
2 
F/U 
3 
F/U 
4 
F/U 
5 
F/U 
6 
STUDY DAY 
-60 
-30 
0 
30 
60 
90 
120 
180 
365 
--- 
--- 
STUDY MONTH 
-1 
0 
--- 
1 
2 
3 
4 
6 
12 
18 
24 
STUDY YEAR 
--- 
--- 
--- 
--- 
--- 
--- 
--- 
--- 
1 
--- 
2 
History1 
X 
 
 
 
 
X 
 
 
 
 
 
Physical Exam2 
X 
 
 
 
 
X 
 
 
X 
 
X 
Pancreatic  cell 
Function3 
X 
 
X 
X 
X 
X 
 
X 
X 
X 
X 
Laboratory 
Studies4 
X 
 
X 
X 
X 
X 
 
 
X 
 
X 
Adverse Event 
Assessment 
X 
 
X 
X 
X 
X 
 
 
 
 
 
Viral Studies5 
X 
 
 
 
 
 
 
 
 
 
 
Survival and 
Disease Status6 
 
 
 
 
 
 
 
 
 
 
 
Serum Pregnancy 
X 
 
 
 
 
 
 
 
 
 
 
Leukapheresis 
 
X 
 
 
 
 
 
 
 
 
 
Potency Assay7 
X 
X 
 
 
 
X 
 
X 
X 
X 
X 
 
 
 
VISIT NAME 
VISIT 
11 
VISIT 
12 
VISIT 
13 
VISIT 
14 
PROCEDURES 
LTFU 1 
LTFU 2 
LTFU 3 
LTFU 4 
STUDY DAY 
--- 
--- 
--- 
--- 
STUDY MONTH 
30 
36 
42 
48 
STUDY YEAR 
--- 
3 
--- 
4 
History1 
 
 
 
 
Physical Exam2 
 
X 
 
X 


 
 
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CONFIDENTIAL 
Pancreatic  cell Function3 
 
X 
 
X 
Laboratory Studies4 
 
X 
 
X 
Adverse Event Assessment 
 
 
 
 
Viral Studies5 
 
 
 
 
Survival and Disease Status6 
X 
X 
X 
X 
Serum Pregnancy 
 
 
 
 
Leukapheresis 
 
 
 
 
Potency Assay7 
 
 
 
 
 
 
1. Medical history including prior treatment, current medicines, and drug allergies 
2. Physical exam including blood pressure, pulse, temperature, weight, and retinal exam 
3. MMTT stimulated C-peptide, glucose tolerance, glycosylated hemoglobin (A1c), insulin, and GAD65 
autoantibody levels 
4. CBC/differential, creatinine, AST, ALT, and total bilirubin; including creatinine clearance and albuminuria 
5. HIV-1, HIV-2, HTLV-1/2, and HCV serology and HBV surface antigen, surface antibody, and core antibody 
6. Can be conducted either in the office or via telephone interview 
7. Blood draw (60-75 mL; 7-8 BD Vacutainer Sodium Heparin (NH), 143 USP UNITS blood collection tube 
(10ml.16x100mm, REF 367874)) and sent to Jiang Lab 
*** If patient is off study, end of study evaluations within 30 days from last vaccination 
 
 
15 Appendices 
 
15.1 Appendix A:  Monitoring Plan 
15.2 Appendix B:  Data Safety Monitoring Board Charter 
15.3 Appendix C:  Combination Informed Consent and HIPPA Form 


 
 
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16 References 
1. 
Jiang, H., and Chess, L. 2009. How the Immune System Achieves Self Non-self Discrimination 
During Adaptive Immunity. Advances in Immunology 102:94-133. 
2. 
Zheng, H., Zhang, H., Han, M., Nie, H., Wu, Y., Sheng, X., Chess, L., and Jiang, H. 2013. A Unified 
"Class Action" Of Self-Nonself Discrimination Mediates Both Neonatal Transplantation Tolerance And 
Control Of Autoimmune Disease. In Submision. 
3. 
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Hayakawa, T., Miao, D., Liu, E., et al. 2003. In vivo expression of B:9-23 peptide/I-A(g7) complex may 
abrogate the inhibition of diabetes induced by RGD-fiber-mutant adenovirus in NOD mice. Ann N Y Acad 
Sci 1005:218-221. 
 
 


 
 
 
 
 
  
DATA AND SAFETY MONITORING PLAN 
 
 
 
A Phase I/II, Double-Blind, Placebo Controlled, Parallel Design of AVOTRES Cell-
Based Therapy as a Treatment for Human Type 1 Diabetes 
 
 
 
 
 
Principal Investigator: 
Daniel S. Donovan, Jr., MD, MS, CDE 
 
 
 
 
Sponsor: 
Hong Jiang, MD, PhD 
Columbia University 
 
 
 
 
July 15, 2013 


 
Data and Safety Monitoring Plan 
 
Page 2 of 7 
 
Protocol Name: A Phase I/II, Double-Blind, Placebo Controlled, Parallel Design of AVOTRES Cell-Based Therapy as a Treatment 
for Human Type 1 Diabetes 
 
Table of Contents 
 
 
1. 
Purpose and Assessment of Risk ......................................................................................................... 3 
2. 
Glossary of Terms ................................................................................................................................ 3 
3. 
Protocol Summary ............................................................................................................................... 4 
4. 
Monitor Selection and Training........................................................................................................... 5 
5. 
Monitoring Process ............................................................................................................................. 5 
5.1 
Frequency...................................................................................................................................... 5 
5.2 
Monitoring Visits ........................................................................................................................... 5 
5.3 
Anticipated Adverse Events .......................................................................................................... 6 
5.4 
Monitoring Reports ....................................................................................................................... 6 
5.5 
Reporting of AEs/SAEs .................................................................................................................. 6 
5.6 
Corrective action ........................................................................................................................... 6 
6. 
Data Management .............................................................................................................................. 6 
7. 
Drug Accountability ............................................................................................................................. 6 
8. 
Close Out Visit ..................................................................................................................................... 7 
 
 


 
Data and Safety Monitoring Plan 
 
Page 3 of 7 
 
Protocol Name: A Phase I/II, Double-Blind, Placebo Controlled, Parallel Design of AVOTRES Cell-Based Therapy as a Treatment 
for Human Type 1 Diabetes 
1. Purpose and Assessment of Risk 
The monitoring of a clinical trial is necessary to ensure the protection of the subject’s rights, the 
safety of subjects enrolled in the trial and the integrity and quality of the resulting data on an on-
going basis for the study duration. This monitoring plan details the Case Report Form (CRF) and 
source data verification of efficacy and safety parameters, the frequency of monitoring visits and 
reporting, regulatory document review, drug accountability and compliance review, and stopping 
rules for the study.  This is a single site clinical trial with subjects being recruited at Columbia 
University Medical Center. Dr Hong Jiang the Regulatory Sponsor for this trial, and will be 
responsible for the conduct of the trial at the site in accordance with FDA requirements.  
 
Monitoring will be conducted according to Good Clinical Practices, applicable federal regulations 
and the applicable Columbia University Standard Operating Procedures. 
 
There is also a Data Safety and Monitoring Board (DSMB) which will monitor and audit the trial for 
compliance at the site. The Regulatory sponsor will also appoint an individual monitor for regulatory 
oversight of the entire trial, as per FDA guidelines for INDs. 
2. Glossary of Terms 
 
Adverse Event (AE) 
Any untoward medical occurrence in a subject or clinical investigation subject administered a 
pharmaceutical product and that does not necessarily have a causal relationship with this 
treatment.  An adverse event (AE) can therefore be any unfavorable and unintended sign (including 
an abnormal laboratory finding), symptom, or disease temporally associated with the use of a 
medicinal (investigational) product, whether or not related to the medicinal (investigational) 
product.  (ICH) 
 
Case Report Form (CRF) 
 
A printed, optical, or electronic document designed to record all of the protocol-required 
information to be reported to the sponsor on each trial subject. (ICH) 
 
Data Safety Monitoring Board (Monitoring Committee, Data Monitoring Committee) 
A Data Safety Monitoring Board that will be established by the sponsor to assess at intervals the 
progress of a clinical trial, the safety data, and the critical efficacy endpoints, and to recommend to 
the sponsor whether to continue, modify, or stop a trial. (ICH) 
 
Informed Consent 
A process by which a subject voluntarily confirms his or her willingness to participate in a particular 
trial, after having been informed of all aspects of the trial that are relevant to the subject’s decision 
to participate.  Informed consent is documented by means of a written, signed, and dated informed 
consent form. (ICH)  Under 21 CFR 50.20, no informed consent may include any “language through 
which the subject or the representative is made to waive or appear to waive any of the subject’s 


 
Data and Safety Monitoring Plan 
 
Page 4 of 7 
 
Protocol Name: A Phase I/II, Double-Blind, Placebo Controlled, Parallel Design of AVOTRES Cell-Based Therapy as a Treatment 
for Human Type 1 Diabetes 
legal rights or releases or appears to release the investigator, the sponsor, the institution, or its 
agents from liability for negligence.” 
 
Monitoring 
The act of overseeing the progress of a clinical trial, and of ensuring that it is conducted, recorded, 
and reported in accordance with the protocol, standard operating procedures (SOPs), Good Clinical 
Practice (GCP), and the applicable regulatory requirement(s). (ICH) 
 
Monitoring Report 
A written report from the monitor to the sponsor after each site visit and/or other trial-related 
communication according to the sponsor’s SOPs. (ICH) 
 
Serious Adverse Event (SAE) 
Any untoward medical occurrence that at any dose:  results in death, is life-threatening, requires 
subject hospitalization or prolongation of existing hospitalization, results in persistent or significant 
disability/incapacity, or is a congenital anomaly/birth defect. (ICH) 
 
Source Data 
All information, original records and certified copies of original records of clinical findings, 
observations, or other activities in a clinical trial necessary for the reconstruction and evaluation of 
the trial.  Source data are contained in source documents (original records of certified copies). (ICH) 
 
Source Documents 
Original documents, data, and records (e.g., hospital records, clinical and office charts, laboratory 
notes, memoranda, subjects’ diaries or evaluation checklists, pharmacy dispensing records, 
recorded data from automated instruments, copies or transcriptions certified after verification as 
being accurate and complete, microfiches, photographic negatives, microfilm or magnetic media, x-
rays, subject files, and records kept at the pharmacy, at the laboratories, and at medico-technical 
departments involved in the clinical trial). (ICH) 
3. Protocol Summary 
Type 1 diabetes (T1D) is a complex, chronic, T cell-mediated autoimmune disease in which the 
insulin producing β cells are destroyed. In T1D, there is a loss of tolerance to pancreatic β cells that 
results in a CD4 and CD8 T cell-dependent autoimmune process that culminates in complete 
destruction of insulin-producing β cells, leading to insulin deficiency and dysregulated glucose 
metabolism. Diabetic patients manage their hyperglycemia by daily insulin injections, however this 
treatment does not cure the disease or prevent the possibility of the disease’s serious effects, which 
may include kidney failure, blindness, nerve damage, heart attack, stroke, and pregnancy 
complications (JDRF website). It is important then that other therapies to treat T1D be identified and 
tested. As discussed below, immunotherapeutic approaches that effectively prevent and or treat 
T1D have been moving into the clinic recently. Over the years, studies using the non-obese diabetic 
(NOD) mouse model have provided important insights into the immunology of T1D and have 
demonstrated the feasibility of preventing or curing T1D in an antigen-specific manner by 
specifically manipulating disease-relevant autoreactive T cell populations without compromising the 
immune system at large. Avotres, represents one such approach to manipulate autoreactive T-cell 


 
Data and Safety Monitoring Plan 
 
Page 5 of 7 
 
Protocol Name: A Phase I/II, Double-Blind, Placebo Controlled, Parallel Design of AVOTRES Cell-Based Therapy as a Treatment 
for Human Type 1 Diabetes 
responses against beta-cell antigens, that we would like to test under this new IND. Modulating the 
activity of self-reactive CD8+ T cells to damper the activation level may be one approach to treat 
T1D.  
 
Our working model for maintenance of T cell self-tolerance holds that a subset of HLA-E restricted 
regulatory CD8+ T cells that specifically recognize the target structure of hHsp60sp associated with 
HLA-E expressed on the pathogenic self-reactive T cells and selectively down-regulate these 
pathogenic T cells which are responsible for autoimmune diseases. The failure of this HLA-E-
restricted CD8+ T cell subset to recognize, or to suppress, its target population results in a 
permissive state in which organ-specific autoimmunity can emerge. We have identified defects in 
this CD8+ T cell system among nine of ten T1D subjects evaluated, and found the defect to be 
reversible in most of these in vitro by co-incubation with hHsp60sp-loaded autologous immature 
dendritic cells (AVOTRES).  
4. Monitor Selection and Training 
The sponsor will designate a qualified individual monitor for regulatory oversight of the entire trial, 
as per FDA guidelines for INDs.   One monitor will be assigned for this trial and will be responsible to 
complete the monitoring process.  The monitor will be selected by the sponsor based on experience 
and qualifications.  The monitor will be qualified both by education and understanding of Good 
Clinical Practice requirements as will as regulatory compliance. 
 
A CV for the monitor will be obtained and updated annually.  The CV will be kept on file in the 
Sponsor section of the Regulatory Binder to document the qualifications of the monitor. 
 
Prior to study initiation, the monitor will be trained on the protocol and case report forms.   
 
5. Monitoring Process 
5.1 
Frequency 
Monitoring will be conducted on the first three subjects enrolled. The frequency of monitoring 
additional subjects will be determined after the review of the first three subjects. The Principal 
Investigator or a member of the study staff must inform the monitor when a subject is enrolled.  
 
5.2 
Monitoring Visits  
Informed consent, enrollment logs, regulatory documentation, subject source data, and case report 
forms will be reviewed 100% for all subjects.  This will be conducted in accordance with applicable 
regulations and University policies 
 
The monitor will perform 100% drug accountability in an ongoing manner at the time of each 
monitoring visit.  The monitor will confirm the receipt, use and return or disposal of drug. 
 


 
Data and Safety Monitoring Plan 
 
Page 6 of 7 
 
Protocol Name: A Phase I/II, Double-Blind, Placebo Controlled, Parallel Design of AVOTRES Cell-Based Therapy as a Treatment 
for Human Type 1 Diabetes 
5.3 
Anticipated Adverse Events  
Side effects of the investigational agent is not completely known.  However, based on previous 
clinical studies of vaccines in T1D patients, vaccines are well tolerated with little to no aide-effects. 
 
Adverse effects associated with the blood draws include: (common, greater than 10%) local pain or 
swelling; (uncommon, 1-10%) vein damage or irritation/phlebitis, excessive bleeding or bruising, 
feeling faint, infections. 
 
5.4 
Monitoring Reports  
Monitoring reports will be provided to the Regulatory Sponsor within two weeks of completion of 
monitoring visit. The Principal Investigator will take action as required.  
 
5.5 
Reporting of AEs/SAEs 
 
The Sponsor will act as the central reporting entity to the FDA. The PI will be responsible for 
contacting the Sponsor within 24 hours of an SAE, and completing both a SAE form and a MedWatch 
Form documenting the event within the following 48 hours. Copies of these forms will be kept on 
file at Columbia University.  The Sponsor will submit to the FDA a 7-day report for any unexpected 
fatal or life-threatening experience associated with use of the investigational agents or a 15 day 
report for AEs that are serious and unexpected associated with the use of the investigational agent 
under Dr. Jiang’s IND. On an annual basis, a summary of all AEs will be provided to the FDA. 
 
5.6 
Corrective action 
 
If non-compliance is determined, the PI will be notified in writing. This notification will include a 
statement of the corrective action that is required to resolve the situation and the consequences to 
be taken or contemplated should the action not be implemented. The PI will have 10 business days 
to respond to the notification, at which time, no additional subjects can be enrolled.  
6. Data Management 
The PI and Research Coordinator will be responsible for CRFs. Clinical and laboratory data  
will be 
entered directly into the CRF by the Research Coordinator. The monitor will perform data queries 
and ensure database corrections.  
 
CRFs are maintained and stored according to GCP. The clinical records are stored according  to 
policies and procedures established Columbia University. 
7. Drug Accountability 
Sponsor and Investigator will maintain drug accountability in accordance with General 
Responsibilities of Sponsors (21 CFR 312.50) and Investigators (21 CFR 312.60).  The monitor will 
perform 100% drug accountability in an ongoing manner at the time of each monitoring visit.  The 
monitor will confirm the receipt, use and return or disposal of drug. 


 
Data and Safety Monitoring Plan 
 
Page 7 of 7 
 
Protocol Name: A Phase I/II, Double-Blind, Placebo Controlled, Parallel Design of AVOTRES Cell-Based Therapy as a Treatment 
for Human Type 1 Diabetes 
8. Close Out Visit 
The monitor will conduct the Close Out Visit within 1 month after the last subject has completed the 
study. 
 
The following activities will be completed by the monitor to close out the study: 
 
Ensure all data has been reviewed and collected; 
 
Ensure all outstanding queries are answered; 
 
Confirm all Serious Adverse Events, MedWatch Reports and IND Safety Reports, if 
applicable, have been reported to the IRB(s), Sponsor and/or FDA; 
 
Review the Regulatory documentation and Subject Files for completeness and compliance 
with GCP and all applicable federal regulations; 
 
Ensure initial and revised 1572 forms were submitted to the IRB (if applicable); 
 
Ensure all protocol violations were submitted to the IRB; 
 
Ensure that all continuing review reports were submitted to the IRB;  
 
Perform drug accountability;  
 
Review requirements for record retention with the investigator and the clinical staff. 
 
A copy of the report will be sent to the Regulatory Sponsor.  
 


  
 
 
AVOTRES DSMB Charter (FINAL.rev1) 
 
 
 
 
 
Page 1 of 6 
Data Safety Monitoring Board Charter 
 
A Phase I/II, Double-Blind, Placebo Controlled, Parallel Design of AVOTRES Cell-
Based Therapy as a Treatment for Human Type 1 Diabetes 
 
Principal Investigators: Daniel S. Donovan Jr. 
 
 
This Data and Safety Monitoring Board (DSMB) Charter describes the Board responsibilities, 
membership, meeting processes, reports and reporting process.  The DSMB will act in an advisory 
capacity to the study sponsor (Hong Jiang, MD, PhD.) and Columbia University Medical Center, to 
monitor patient safety of the intervention of the above-noted study. Dr. Daniel Donovan is the 
investigator conducting the above-noted clinical trial. 
 
Reports will be provided to the DSMB by the investigative staff.  
DSMB RESPONSIBILTIES 
The initial responsibility of the DSMB will be to review the research protocol, informed consent 
documents and plans for data safety and monitoring, and to approve the initiation of this clinical trial if 
all the above are satisfactory. After this approval, and at periodic intervals (described below) during the 
course of the trial, the DSMB responsibilities are to: 
 
evaluate the progress of the trial, including periodic assessments of data quality and timeliness, 
participant recruitment, accrual and retention, participant risk versus benefit, aspects of trial 
conduct, and other factors that can affect study outcome; 
 
consider factors external to the study when relevant information becomes available, such as 
scientific or therapeutic developments that may have an impact on the safety of the participants 
or the scientific value or ethics of the trial; 
 
ensure that the safety of the study participants is not being compromised; 
 
make recommendations to the sponsor, the PI, the Safety Advisor, the institution, and 
Institutional Review Board (IRB) concerning continuation, termination or other modifications of 
the trial based on the observed beneficial or adverse effects of the treatment under study 
 
consider whether the data exceed any early stopping boundaries which have been clearly 
defined in advance of data analysis; 
 
maintain the confidentiality of the trial data and the results of monitoring; and, 
 
assist the sponsor by making recommendations regarding any problems with study conduct, 
enrollment, and sample size and/or data collection.  
MEMBERSHIP 
 
The DSMB will consist of five voting members. The DSMB voting members have no financial, 
scientific, or other conflict of interest with the trial.  The DSMB coordinator is [TBD] and is 
employed by [TBD]. The role of the DSMB coordinator is to schedule and coordinate the 
meetings, assure a quorum is constituted and provide minutes of the meeting. The members 
have been selected by Dr. Jiang, the study sponsor. The DSMB includes members with expertise 


  
 
 
AVOTRES DSMB Charter (FINAL.rev1) 
 
 
 
 
 
Page 2 of 6 
Data Safety Monitoring Board Charter 
in the clinical area under study and/or clinical trial methodology.  All voting members will 
provide written documentation regarding absence of any disqualifying conflicts of interest.  
[TBD] has been selected by the sponsor to serve as the Chairperson. S/he is responsible for 
overseeing the meetings and developing the agenda in consultation with the non-voting 
coordinating member and the PI.  [TBD] will serve as the liaison between the study sponsor and 
investigator, and the DSMB, and will provide the logistical management for and support of the 
DSMB.  
 
Contact Information 
 
Name 
Role 
Contact Information 
Hong Jiang, MD, PhD 
Sponsor 
Columbia University Medical Center 
PH - 8 East, Ste. 101,  
622 West 168th St - New York, NY 10032 
Phone: 212-305-9984/201-944-3352     
Fax: 212-305-6070  
Email: hj4@columbia.edu 
Daniel Donovan, MD, 
MS 
Principal Investigator 
Columbia University Medical Center 
161 Fort Washington Ave. Room 2-212 
Phone: 212-305-0504 
Email: dsd8@columbia.edu 
TBD 
Clinical Trials Coordinator 
TBD 
TBD 
DSMB Coordinator 
TBD 
TBD 
Chair, and Voting DSMB 
Member 
TBD 
TBD 
Voting DSMB Member 
TBD 
TBD 
Voting DSMB Member 
 
TBD 
TBD 
Voting DSMB Member 
TBD 
TBD 
Voting DSMB Member 
TBD 
Rajasekhar 
Ramakrishnan, 
Eng.Sc.D. 
 
Study Statistician 
Director, Biomathematics Division 
Department of Pediatrics  
Address: 601 West 168 Street #54 
New York NY 10032 
Phone: 212 305 7989  
Fax: 212 305 5834  
Email: rr6@columbia.edu  
BOARD PROCESS  
The first meeting of the DSMB will be devoted to a discussion of the protocol and associated documents.  
The DSMB will consider whether any modifications of the trial would be desirable, discuss guidelines for 
monitoring the study, and discuss, finalize and approve the DSMB charter. The Sponsor (or designated 
representative), will prepare the agenda to address the review of the manual of operating procedures, 


  
 
 
AVOTRES DSMB Charter (FINAL.rev1) 
 
 
 
 
 
Page 3 of 6 
Data Safety Monitoring Board Charter 
modification of the study design, initiation of the trial, monitoring roles, reporting of adverse events, 
stopping rules, and interim analysis plan.   
 
The DSMB meeting schedule will be based on safety events and study milestones. The DSMB will have 
an initial meeting prior to study initiation.  Subsequent meetings may occur via teleconference after the 
first three participants complete his/her three vaccinations. If necessary, additional meetings of the 
Board may be held if there are safety issues between scheduled meetings.  Meetings will also be held at 
a minimum of every six months.  Meetings may be closed at the committee’s request.  The initial and 
subsequent meetings may occur either via teleconference or face-to-face. 
 
The DSMB will convene at any time during the study in the event of a protocol specific, study related 
serious adverse event that is considered to be possibly, likely or definitely related to treatment. 
 
Meetings shall be closed to the public because discussions may address confidential patient data. The 
principal investigator and members of his staff may attend portions of the meeting as appropriate, but 
the DSMB may conduct some sessions without presence of the sponsor or study personnel.   
MEETING FORMAT AND FREQUENCY 
An appropriate format for DSMB meetings consists of an open and a closed session. The open sessions 
may be attended by the principal investigator(s), and institution staff.  Issues discussed at open sessions 
will include conduct and progress of the study, including patient accrual, compliance with protocol, and 
problems encountered. Closed sessions will be attended only by voting members and will include 
discussion of safety data.  The DSMB will vote on the continuation or termination of the study during the 
closed session. 
 
At any time, the committee may request an executive closed session.   
 
A minimum of four (4) voting members of the DSMB will be required to determine continuance or 
discontinuance of the study. In the event of a split vote (to be defined by the DSMB), study enrollment 
will be interrupted but participants already enrolled and dosed will continue to be closely followed.    
The sponsor will immediately notify OHR, and the IRB with copies to the FDA.  In such a case, the 
sponsor, and FDA will need to decide the final disposition of the study 
 
The DSMB will provide advice to the investigator, and will consult with Sponsor as necessary. The initial 
responsibility of the DSMB will be to approve the initiation of this clinical trial. At later meetings, the 
DSMB will evaluate patient-subject safety and vote on continuation of the study at the following study 
milestones: 
 
 
 
 
If necessary, additional meeting of the DSMB may be held if safety issues arise between scheduled 
meetings. The DSMB's findings and recommendations regarding patient safety and continuation or 
termination of the trial will be shared with the Sponsor, Columbia University IRB, and FDA.  


  
 
 
AVOTRES DSMB Charter (FINAL.rev1) 
 
 
 
 
 
Page 4 of 6 
Data Safety Monitoring Board Charter 
 
PROTOCOL STOPPING RULES (same for all protocols) 
The study will be stopped if: 
 
Premature study termination may occur if the Investigator, Study Funder, Sponsor, DSMB or any 
independent review board or regulatory body decides for any reason that subject safety may be 
compromised by continuing the study. 
 
Premature study termination may occur if the Sponsor or Study Funder decides to discontinue 
the development of the intervention to be used in this study. 
The protocol will also be stopped if a third or more of the patients have ≥ grade 3 toxicity as determined 
by CTC, which is unmanageable, unexpected and unrelated, and attributable to protocol therapy. 
Depending on the nature of the toxicities, the protocol may be amended to lower the AVOTRES dose or 
make other modifications that would limit toxicities observed.  Dose limiting toxicity (DLT) is defined as a 
≥ grade 3 toxicity as determined by CTC, which is unmanageable, unexpected and unrelated to 
chemotherapy and attributable to protocol therapy.  
The safety assessment will be made after completion of the first three subjects enrolled and again after 
the second three (subjects 4-6) subjects are enrolled.  In the event of DLT in the first 6 subjects, the 
study will continue or be stopped in accordance with the following: 
 
1. The study will continue if < 2 out of the first 3 patients experience DLT; the study will stop if  2 or 
more in the first 3 patients experience DLT. Based on this rule, if 1 of 3 subjects experiences a DLT, 
the 95% CI for the true toxicity rate is 0.8% to 91%. The probability to observed 1 of 3 subjects 
experiences a DLT is <44% if the true DLT rate is >33%, and 
2. The study will continue if <3 out of the first 6 patients experience DLT; the study will stop if 3 or 
more in the first six patients experience a DLT.  Based on this rule, if 2 of 6 subjects experience a 
DLT, the 95% CI for the true toxicity rate is 4% to 78%. The probability to observed 2 of 6 subjects 
experiences a DLT is <33% if the true DLT rate is >33%. 
If the study continues enrollment after 6 subjects (i.e. DLT is observed in fewer than 3 of 6 subjects), the 
study will be stopped in the event that a total of three subjects in the study have experienced a DLT. 
The study will be paused if:  
 
The protocol will be paused pending review by IRB, FDA and the DSMB if any patient experiences 
any of the following events within two weeks of the CART-19 infusion   
o respiratory failure requiring mechanical ventilation; 
o any adverse event, exclusive of elective surgical procedures, requiring admission to an 
intensive care unit for management;   
o life-threatening (grade 4) toxicity attributable to protocol therapy; or 
o death. 
REPORTS  
1. Immediate response 
In the event of a decision to close the study, the Principal Investigator, study sponsor, and IRB 
will be immediately notified.  FDA will be subsequently notified in writing as soon as possible. 
 


  
 
 
AVOTRES DSMB Charter (FINAL.rev1) 
 
 
 
 
 
Page 5 of 6 
Data Safety Monitoring Board Charter 
2. Serious Adverse Event Reporting 
In the event of a Serious Adverse Event as defined in the protocol Section 10 that is considered 
to be possibly, likely or definitely related to treatment, the following process will occur.  The 
principal investigator will notify the IRB and the study sponsor.  The research coordinator/nurse 
will fax a completed Serious Adverse Event form to the Sponsor.  The IRB will be notified via 
RASCAL.  A phone call and/or e-mail will also be sent to the Sponsor.  The sponsor will notify the 
FDA.  The DSMB Coordinator will notify the DSMB and convene an immediate teleconference 
with the chair and board.   The outcome of the DSMB review and discussion of the SAE will be 
provided to the sponsor who will notify the FDA, and will assure that the IRB is notified of the 
outcome.  
 
3. Interim Reports: Interim reports will be prepared and distributed to the DSMB at least 10 
working days prior to the scheduled meeting if at all possible. These interim reports are 
numbered and provided by email unless otherwise instructed by the DSMB Chair. The format 
and content of the report will be proposed by the sponsor and may be modified by request of 
the DSMB.   
 
 
Proposed data to be included in the report are as follows: 
 
Summary of Previous Meeting 
 
Provided by DSMB Coordinator 
 
Executive Summary 
 
Enrollment and important findings as determined by Sponsor and PI 
 
Demographics, Study Status and Manufacturing Summary (Includes data on all enrolled 
subjects and any available data on screened but not enrolled subjects) 
 Enrollment (total screened,  total enrolled, pending enrollment, total active) 
 Demographics by subject, no totals or ranges   
 Overview of subjects enrolled (last visit, date of visit, infusion date, all protocol 
deviations, AEs including laboratory AEs [yes/no]) 
 Cell manufacturing and release by subject All Protocol Deviations by subject 
(approved by sponsor) 
 
Data Summaries (The following tables include data from enrolled subjects only) 
 Overview and table/figure of potency assay 
 Overview and table/figure of patient response   
 Summary of Adverse Events (includes laboratory toxicities) 
o Table of adverse events by body system and severity 
o Table of adverse events by body system and relatedness 
o Listings of adverse events by subject (includes AE term, severity and 
relationship) 
 Table of other concomitant medications by patient with start/stop dates 
 
 
4. Reports from the DSMB:  
As previously stated, the formal DSMB report will conclude with a recommendation to continue 
or to terminate or pause the study.  A termination or pause recommendation may be made by 
the DSMB at any time. The DSMB Chair is responsible for notifying the sponsor, and the PI if the 
decision is to terminate or pause.  In the event of a split vote in favor of continuation, a report 


  
 
 
AVOTRES DSMB Charter (FINAL.rev1) 
 
 
 
 
 
Page 6 of 6 
Data Safety Monitoring Board Charter 
will be sent to the IRB with copies to the FDA.  In such a case, the sponsor and FDA will need to 
decide the final disposition of the study.   
 
5. A formal report containing the meeting minutes and any recommendations for continuation or 
modifications of the study will be prepared by the DSMB Coordinator in consultation with the 
DSMB Chairperson.  As noted above, sponsor will have communicated immediate meeting 
outcome to required entities.  This report will be sent to the full DSMB within 2 weeks of the 
meeting. Once approved by the DSMB, the OHR representative will forward the formal DSMB 
recommendation report to the sponsor and PI within 3 weeks of each meeting. It is the 
responsibility of the sponsor and PI to distribute the formal DSMB recommendation report to 
the UPENN IRB.   
CONFIDENTIALITY  
All materials, discussions and proceedings of the DSMB are completely confidential. Members and other 
participants in DSMB meetings are expected to maintain confidentiality.  


AVOTRES: Autologous Dendritic cells loaded with Hsp60sp peptide 
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Appendix C AVOTRES IC and HIPAA Form (FINAL.rev1) 
  
 
COLUMBIA UNIVERSITY 
RESEARCH SUBJECT  
INFORMED CONSENT AND  
HIPAA AUTHORIZATION FORM 
 
 
Protocol Title:   
A Phase I/II Double-Blind, Placebo Controlled, Parallel 
Design of AVOTRES Cell-Based Therapy as a Treatment 
for Human Type 1 Diabetes 
Principal 
Investigator: 
Daniel S. Donovan, Jr., MD, MS, CDE 
Columbia University Medical Center 
161 Fort Washington Ave. Room 2-212 
New York, NY 10032 
Phone: 212-305-0504 
Email: dsd8@columbia.edu 
Emergency 
Contact: 
Name: Leonard Chess MD  
Address: 630 W.168th Street PH 8-east Ste 101 
Phone: 212-305-9984 
Email: lc19@columbia.edu 
 
  
 
 
Why am I being asked to volunteer? 
 
You are being invited to participate in a research study. Your participation is voluntary 
which means you can choose whether or not you want to participate. If you choose not 
to participate, there will be no loss of benefits to which you are otherwise entitled. 
Before you can make your decision, you will need to know what the study is about, the 
possible risks and benefits of being in this study, and what you will have to do in this 
study. The research team is going to talk to you about the research study, and they will 
give you this consent form to read. You may also decide to discuss it with your family, 
friends, or family doctor. You may find some of the medical language difficult to 
understand. Please ask the study doctor and/or the research team about this form. If 
you decide to participate, you will be asked to sign this form. 
 
 


AVOTRES: Autologous Dendritic cells loaded with Hsp60sp peptide 
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Appendix C AVOTRES IC and HIPAA Form (FINAL.rev1) 
  
 
 
What is the purpose of this research study? 
 
We are doing this research study to ﬁnd out if AVOTRES works as a vaccine and can 
correct Type 1 diabetes. 
 
AVOTRES is an investigational drug. This means that the drug has not been approved by 
the Food and Drug Administration (FDA) for medical use in patients, but has only been 
approved for use in research.  
 
You are being asked to take part in this study because you have type 1 diabetes. There 
are 21 people who will be enrolled in this study.  
 
 
How long will I be in the study?  
 
This study takes a total of four (4) years to finish.  You will need to schedule a visit with 
the study physician to see if you can be on this study.  The study outlines special 
circumstances, called Inclusion and Exclusion Criteria, that help the doctor determine if 
you can be on the study.  In order for the doctor to determine if you can be on the 
study, the doctor will need your consent to draw blood and conduct a physical and 
retinal exam.   
 
If you can be included in this study, then you will need to come to the doctor’s office 
many times over the next 6 months.  After 6 months, the visits slow down.  After two 
years, you can speak to your doctor (or another person from the study team) over the 
telephone. 
 
What am I being asked to do? 
 
You are being asked to participate in a study which will see if the AVOTRES vaccine can 
reverse type 1 diabetes.   
 
Fourteen (14) patients will receive the vaccine.  Seven (7) patients will receive saline 
(type of water) only.  This is called randomization, where the patient is randomly given 
AVOTRES or saline.  You will not know which you are getting.   
 
 
What are the possible risks or discomforts?  
 
WE do not know of any risks associated with AVOTRES.  Other studies that looked at 
something similar to AVOTRES had no complications. 
 


AVOTRES: Autologous Dendritic cells loaded with Hsp60sp peptide 
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Appendix C AVOTRES IC and HIPAA Form (FINAL.rev1) 
  
 
You will have a procedure done called APHERESIS, where your blood is taken from one 
arm, filtered, and then put back in your other arm at the same time.  The most common 
side effects are dizziness, mild chest tightness, or cough related to the reinfusion of your 
blood. 
 
Reproductive risks:  Because of the effects of this drug/device, there could be harm to 
unborn children or children who are breast-feeding. These effects could also harm the 
mother.  It is also possible that harmful side effects that are not yet known could 
happen to both the mother and unborn or breast-feeding child.  If you are currently 
pregnant, it is important that you inform the investigator because you will not be able 
participate in the study.  If you are able to become pregnant, you will be given a serum 
pregnancy test before entry into the study.  You are asked to use a medically accepted 
method of birth control while you participate in the study.  You should not become 
pregnant while you are taking AVOTRES.  If you do become pregnant, you must tell the 
investigator and consult an obstetrician or maternal-fetal specialist. 
 
What if new information becomes available about the study? 
During the course of this study, we may find more information that could be important 
to you.  This includes information that, once learned, might cause you to change your 
mind about being in the study.  We will notify you as soon as possible if such 
information becomes available. 
 
What are the possible benefits of the study?  
 
This is the first time that AVOTRES is being tested in human beings.  However, because 
this is the first time AVOTRES is being tested in humans, there is a chance you may not 
get any benefit from being in this research study. 
 
 
What other choices do I have if I do not participate?  
 
You may investigate other ongoing studies and you can discuss alternatives with your 
physician. 
 
Will I be paid for being in this study? 
 
You will not be paid for being in this study. 
 


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Appendix C AVOTRES IC and HIPAA Form (FINAL.rev1) 
  
 
Will I have to pay for anything? 
 
You are still responsible for any deductibles or applicable co-pays for routine office 
visits, scans and blood work.  Please talk to your doctor and study team about putting 
you in touch with a financial counselor to determine exactly what the deductible and co-
pay will be for you; this is highly variable depending on your type of insurance. 
 
 
What happens if I am injured from being in the study?  
 
We will offer you the care needed to treat injuries directly resulting from taking part in 
this research.  We may bill your insurance company or other third parties, if 
appropriate, for the costs of the care you get for the injury, but you may also be 
responsible for some of them. 
 
There are no plans for the Columbia University to pay you or give you other 
compensation for the injury.  You do not give up your legal rights by signing this form.   
 
If you think you have been injured as a result of taking part in this research study, tell 
the person in charge of the research study as soon as possible.  The researcher’s name 
and phone number are listed in the consent form. 
 
 
When is the Study over?  Can I leave the Study before it ends? 
 
This study is expected to end after all participants have completed all visits, and all 
information has been collected.  This study may also be stopped at any time by your 
physician, the study Sponsor, or the Food and Drug Administration (FDA) without your 
consent because: 
 The Primary Investigator feels it is necessary for your health or safety.  Such an 
action would not require your consent, but you will be informed if such a 
decision is made and the reason for this decision. 
 You have not followed study instructions.  
 The Sponsor, the study Principal Investigator, or the Food and Drug 
Administration (FDA) has decided to stop the study. 
 
If you decide to participate, you are free to leave the study at anytime.  Withdrawal will 
not interfere with your future care.   
 
What information about me may be collected, used or shared 
with others?   
 
You will be assigned a unique number that tells the physician and the study team who 
you are, but no one else will know who you are.   


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Appendix C AVOTRES IC and HIPAA Form (FINAL.rev1) 
  
 
 
Why is my information being used? 
 
Your information is used by the research team to contact you during the study. Your information 
and results of tests and procedures are used to: 
 do the research 
 oversee the research 
 to see if the research was done right.   
 
Who may use and share information about me?   
 
The following individuals may use or share your information for this research study: 
 The investigator for the study and the study team  
 Other authorized personnel at Columbia University  
 
Who, outside of Columbia University, might receive my 
information?  
 
 Those working under the direction of the investigator for the study, (e.g. under 
subcontracts). 
 The funding sponsor and organizations supporting the sponsor  
 
Oversight organizations: 
 The Food and Drug Administration 
 The Office of Human Research Protections 
 The study data and safety monitoring board 
 
The Principal Investigator or study staff will inform you if there are any additions to the 
list above during your active participation in the trial. Any additions will be subject to 
Columbia University procedures developed to protect your privacy. 
 
How long may Columbia University use or disclose my personal 
health information?   
 
Your authorization for use of your personal health information for this specific study 
does not expire.  
 
Your information may be held in a research database.  However, the School of Medicine 
may not re-use or re-disclose information collected in this study for a purpose other 
than this study unless: 
 You have given written authorization  
 The Columbia University’s Institutional Review Board grants permission 
 As permitted by law   
 


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Appendix C AVOTRES IC and HIPAA Form (FINAL.rev1) 
  
 
Can I change my mind about giving permission for use of my 
information? 
 
Yes. You may withdraw or take away your permission to use and disclose your health 
information at any time.  You do this by sending written notice to the investigator for the study.  
If you withdraw your permission, you will not be able to stay in this study. 
 
What if I decide not to give permission to use and give out my 
health information? 
 
Then you will not be able to be in this research study. 
 
You will be given a copy of this Research Subject HIPAA Authorization describing your 
confidentiality and privacy rights for this study.  
 
By signing this document you are permitting the School of Medicine to use and disclose personal 
health information collected about you for research purposes as described above. 
 
Who can see or use my information?  How will my personal 
information be protected?   
 
We will do our best to make sure that the personal information obtained during the 
course of this research study will be kept private.  However, we cannot guarantee total 
privacy.  Your personal information may be given out if required by law.  If information 
from this study is published or presented at scientific meetings, your name and other 
personal information will not be used. If this study is being overseen by the Food and 
Drug Administration (FDA), they may review your research records.   
       
Who can I call with questions, complaints or if I’m concerned 
about my rights as a research subject? 
 
If you have questions, concerns or complaints regarding your participation in this 
research study or if you have any questions about your rights as a research subject, you 
should speak with the Principal Investigator listed on page one of this form.   
 
 
 
 
 
 
 


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Appendix C AVOTRES IC and HIPAA Form (FINAL.rev1) 
  
 
 
When you sign this form, you are agreeing to take part in this research study. This 
means that you have read the consent form, your questions have been answered, and 
you have decided to volunteer.  Your signature also means that you are permitting the 
Columbia University to use your personal health information collected about you for 
research purposes within our institution. You are also allowing the Columbia University 
to disclose that personal health information to outside organizations or people involved 
with the operations of this study. 
 
A copy of this consent form will be given to you. 
 
 
________________________       ____________________________________ 
Name of Subject (Please Print)  
Signature of Subject   
           Date 
 
 
 
 
 
________________________ 
_____________________________________ 
Name of Person Obtaining 
 
Signature                                   
Date 
Consent (Please Print) 
 
 


Amendment 3 for Protocol AVT001-T1D-01 
1Feb2021 
AVT001:  Hsp60sp-loaded Autologous Immature Dendritic Cells 
Avotres, Inc. 
Page 1 of 50 
Investigational Product: 
AVT001 Suspension for Intravenous Infusion  
Hsp60sp-loaded Autologous Immature Dendritic Cells 
(7x106-10x106 cells/dose) 
Study Title: 
A Phase 1 / 2 Double-Blind, Randomized, Placebo Controlled 
Study of Safety, Tolerability and Potential Efficacy of 
AVOTRES Cell-Based Therapy (AVT001) in Patients with 
Type 1 Diabetes
Sponsor:
Avotres, Inc. 
140 East Hanover Avenue 
Cedar Knolls, NJ 07927 
Phone: 973-487-6972 
Protocol Number: AVT001-T1D-01 
Initial protocol version:  July 15, 2013 
Amendment 1: February 14, 2019 
Amendment 2: January 22, 2020 
Amendment 3: February 1, 2021 


Amendment 3 for Protocol AVT001-T1D-01 
 
1Feb2021 
AVT001:  Hsp60sp-loaded Autologous Immature Dendritic Cells 
Avotres, Inc. 
Page 2 of 50 
 
1 
Table of Contents 
1 
Table of Contents ........................................................................................................................ 2 
2 
List of Abbreviations .................................................................................................................. 5 
3 
Synopsis........................................................................................................................................ 7 
4 
Introduction ............................................................................................................................... 12 
4.1 
Background .................................................................................................................................................... 12 
4.2 
Failure of self-nonself discrimination by the immune system results in pathogenesis of autotimmune 
disease such as T1D ........................................................................................................................................ 13 
4.3 
The Autoimmune Process of Type 1 diabetes .............................................................................................. 13 
4.4 
Autoreactive T cells in T1D ........................................................................................................................... 14 
4.5 
Immunotherapeutic approaches to treat T1D ............................................................................................. 14 
4.6 
Dendritic cell based therapies for T1D ......................................................................................................... 15 
4.7 
Dosing Rationale ............................................................................................................................................ 15 
5 
Study Objectives and Design ................................................................................................... 18 
5.1 
Primary Outcomes ......................................................................................................................................... 18 
5.2 
Secondary Outcomes ...................................................................................................................................... 18 
5.3 
General Study Design .................................................................................................................................... 18 
5.4 
Study Duration ............................................................................................................................................... 20 
5.5 
Subject Enrollment ........................................................................................................................................ 20 
5.6 
Data Safety Monitoring Board (DSMB) ...................................................................................................... 20 
5.6.1 
Stopping Rules ................................................................................................................................................. 20 
5.7 
Study Endpoints ............................................................................................................................................. 21 
6 
Subject Selection and Withdrawal .......................................................................................... 22 
6.1 
Inclusion Criteria ........................................................................................................................................... 22 
6.2 
Exclusion Criteria .......................................................................................................................................... 22 
6.3 
Early Withdrawal of Subjects ....................................................................................................................... 23 
7 
Description of Study Drug ....................................................................................................... 24 
7.1 
Administration of Study Drug. ...................................................................................................................... 24 
7.2 
Reconciliation of Drug  Product ..................................................................................................................... 25 
8 
Study Procedures ...................................................................................................................... 26 
8.1 
Informed Consent .......................................................................................................................................... 26 
8.2 
Screening/Randomization/Leukapheresis .................................................................................................... 26 
8.2.1 
Screening Study Day ≤ - 90 ............................................................................................................................. 26 
8.2.2 
Randomization ................................................................................................................................................. 27 
8.2.3 
Leukapheresis .................................................................................................................................................. 27 
8.3 
Double-Blind Treatment Period ................................................................................................................... 27 
8.3.1 
Study Days 1, 30 ± 7 days, and 60 ± 7 days .................................................................................................... 27 
8.3.2 
Study Day 90 ± 7 days ..................................................................................................................................... 28 
8.4 
Long-Term Follow-Up Period ....................................................................................................................... 28 
8.4.1 
Study Day 150 ± 7 days ................................................................................................................................... 29 
8.4.2 
Study Day 270 ± 14 days, 360 ± 14 days, 540 ± 14 days, and 720/Early Term ± 14 days .............................. 29 
8.5 
ECGs ............................................................................................................................................................... 29 


Amendment 3 for Protocol AVT001-T1D-01 
 
1Feb2021 
AVT001:  Hsp60sp-loaded Autologous Immature Dendritic Cells 
Avotres, Inc. 
Page 3 of 50 
 
8.6 
MMTT Description ........................................................................................................................................ 29 
8.7 
Prior and Concomitant Therapy and Insulin Usage ................................................................................... 30 
8.8 
Immunoassessment “Potency” Assay Procedure ........................................................................................ 30 
9 
Statistical Methodology ............................................................................................................ 31 
9.1 
Analysis Timing and Blinding Considerations ............................................................................................ 31 
9.2 
Sample Size Determination ........................................................................................................................... 31 
9.3 
Analysis Populations ...................................................................................................................................... 31 
9.4 
Baseline characteristics, Patient Disposition and Medication .................................................................... 31 
9.5 
Adverse Events ............................................................................................................................................... 31 
9.6 
Severe Hypoglycemic Events and Insulin Log ............................................................................................. 32 
9.7 
Clinical Laboratories, ECGs, and Vital Signs ............................................................................................. 32 
9.8 
Pharmacodynamic Outcomes ....................................................................................................................... 32 
9.8.1 
CD8+ T-cell reg system ................................................................................................................................... 32 
9.8.2 
C-Peptide AUC Calculation ............................................................................................................................. 32 
9.8.3 
HbA1c, Antibodies .......................................................................................................................................... 33 
10 
Risks and Discomforts .............................................................................................................. 34 
10.1 
Venipuncture .................................................................................................................................................. 34 
10.2 
Leukapheresis................................................................................................................................................. 34 
10.3 
Response to AVT001 ....................................................................................................................................... 34 
11 
Safety and Adverse Events ....................................................................................................... 35 
11.1 
Definitions ....................................................................................................................................................... 35 
11.1.1 
Unanticipated Problems Involving Risk to Subjects ........................................................................................ 35 
11.1.2 
Adverse Event .................................................................................................................................................. 35 
11.1.3 
Serious Adverse Event (SAE) .......................................................................................................................... 36 
11.1.4 
General Physical Examination Findings .......................................................................................................... 37 
11.1.5 
Post-study Adverse Event ................................................................................................................................ 37 
11.1.6 
Abnormal Laboratory Values........................................................................................................................... 37 
11.1.7 
Severe Hypoglycemia ...................................................................................................................................... 38 
11.2 
Recording of Adverse Events ........................................................................................................................ 38 
11.3 
Reporting of Adverse Events ......................................................................................................................... 38 
11.3.1 
Adverse Event Reporting Period ...................................................................................................................... 38 
11.3.2 
IRB Notification .............................................................................................................................................. 39 
11.3.3 
FDA Notification by Sponsor .......................................................................................................................... 40 
11.4 
Protocol Deviations ........................................................................................................................................ 41 
11.5 
Medical Monitoring ....................................................................................................................................... 41 
12 
Data Handling and Record Keeping ....................................................................................... 42 
12.1 
Confidentiality ................................................................................................................................................ 42 
12.2 
Source Documents .......................................................................................................................................... 42 
12.3 
Case Report Forms ........................................................................................................................................ 42 
12.4 
Records Retention .......................................................................................................................................... 42 
13 
Study Monitoring, Auditing, and Inspecting .......................................................................... 43 
13.1 
Study Monitoring Plan .................................................................................................................................. 43 
13.2 
Auditing and Inspecting ................................................................................................................................ 43 


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14 
Ethical Considerations .............................................................................................................. 44 
15 
References .................................................................................................................................. 46 
16 
Declarations of Sponsor and Investigator .............................................................................. 50 
 
 


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2 
List of Abbreviations 
Abbreviation
Definition 
A1c 
Hemoglobin A1c 
APC 
Antigen Presenting Cells 
ATG 
Anti-thymocyte Globulin 
CBC 
Complete Blood Count 
CD4, CD8 
Cluster of Differentiation 
CFR 
Code of Federal Regulations 
CMP 
Complete Metabolic Panel 
CRF 
Case Report Form 
CTCAE 
Common Terminology Criteria for Adverse Events 
CTL 
Cytotoxic T Cell 
DC 
Dendritic Cell 
DFCI-CMCF
Dana Farber Cancer Institute, Cell Manipulation Core Facility 
DIFF 
Differential 
DMSO 
Dimethyl sulfoxide 
DMK 
Dystrophia Myotonica Kinase 
DLT 
Dose Limiting Toxicity 
DSMB 
Data Safety Monitoring Board 
GAD 
Glutamic Acid Decarboxylase 
GCP 
Good Clinical Practice 
GFAP 
Glial Fibrillary Acidic Protein 
GM-CSF 
Granulocyte Macrophage-Colony Stimulating Factor 
HbA1c 
Hemoglobin A1c 
HBsAg 
Hepatitis B Virus Surface Antigen 
HBV 
Hepatitis B Virus 
HCV 
Hepatitis C Virus 
HIV 
Human Immunodeficiency Virus 
HLA 
Human Leukocyte Antigen 
Hsp60sp 
Heat Shock Protein 60 Signal Peptide 
ICA 
Islet Cell Antibody 
iDC 
Immature Dendritic Cell 
IFNγ 
Interferon γ 
IGRP 
Islet-Specific Glucose-6-Phosphate Catalytic Subunit Related Protein


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IL 
Interleukin 
IP 
Investigational Product 
i.v. 
Intravenous 
mcg 
Micrograms 
MHC 
Major Histocompatability Complex 
MMRM 
Mixed model for repeated measures 
MMTT 
Mixed Meal Tolerance Test 
MOP 
Manual of Operating Procedures 
MRI 
Magnetic Resonance Image 
mRNA 
Messanger Ribonucleic Acid 
NOD 
Non-Obese Diabetic (murine model) 
ppIAPP 
Pre-Pre-Islet Amyloid Polypeptide Protein 
Qa-1 
Murine MHC class I molecule 
RNA 
ribonucleic acid 
SAP 
Statistical analysis plan 
s.c. 
Subcutaneous 
T1D 
Type 1 diabetes 
TNFα 
Tumour Necrosis Factor α 
Treg cells 
Regulatory T Cells 
 


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3 
Synopsis 
Name of Sponsor/Company: 
Avotres, Inc. 
Name of Investigational Product (IP): 
AVT001 suspension for intravenous infusion (AVT001) 
Name of Active Ingredient: 
The study drug represents an individualized preparation of autologous immature dendritic cells, derived 
by culture from the subject’s adherent primary monocytes with GM-CSF and IL-4 for 6 days, and 
loaded passively with a synthetic oligopeptide from Hsp60sp. 
Title of Study:  
A Phase 1 / 2 Double-Blind, Randomized, Placebo Controlled Study of Safety, Tolerability and 
Potential Efficacy of AVOTRES Cell-Based Therapy (AVT001) in Patients with Type 1 diabetes 
Study center(s):  This study is planned as a single-center study, at the Joslin Diabetes Center.  
However, an additional center may be added as enrollment and accrual needs dictate. 
Principal Investigator:  
Jason Gaglia, MD, MMSc 
Joslin Diabetes Center  
1 Joslin Place, Room 478 
Boston, MA 02215 
Admin: 617-309-4024 
Direct: 617-309-4214 
Fax: 617-309-4310 
Email: Jason.Gaglia@joslin.harvard.edu 
Studied period (years):  
Duration of the study is approximately 3 years, from 
enrollment of the first subject until last subject completes 2 
years of follow up. 
Phase of development:  
Phase 1 / 2 
Objectives:  
Primary Objectives: 
 
To determine the safety and tolerability profile of three doses of AVT001 (as compared to 
placebo) in T1D subjects with in vitro-reversible defects in the CD8+ T cell regulatory 
system 
Secondary Objectives 
 
To determine whether, among T1D subjects with in vitro-reversible defects in the CD8+ T 
cell regulatory system, infusion of AVT001 will lead to an in vivo reconstitution of this 
system. 
 
To determine whether, among T1D subjects with in vitro-reversible defects in the CD8+ T 
cell regulatory system, infusion of AVT001 will result in  cell preservation assessed by 
stimulated C-peptide. 
Methodology:  
This is a double-blind, randomized, placebo-controlled study to evaluate the safety and tolerability of 
AVT001, and to assess AVT001 as a potential treatment for type 1 diabetes (T1D).  The trial will 
involve approximately 24 new-onset T1D subjects who have been identified as having: (a) a defect in 


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HLA-E-restricted CD8+ T cell function associated with pancreatic β cell destruction; and (b) evidence 
that this defect of HLA-E-restricted CD8+ T cells can be corrected by in vitro exposure to the 
proprietary AVT001 procedure.  
Each subject will be randomized to one of two treatment groups: 
 
AVT001:  16 subjects to receive AVT001 through i.v. administration.   
 
Placebo control:  8 subjects to receive placebo infusion solution only through i.v. 
administration.   
Subjects will be treated with 3 consecutive treatments, administered with one month (+/-7 days) 
intervals between doses. Infusions must be at least 21 days apart. The primary time point for assessment 
and statistical analysis will be 3 months post-last dose (Month 5), with longer-term follow-up through 
22 months post-last dose (Month 24). 
This is a first-in-man study, and therefore, exposure will be managed using a staggered enrollment that 
will allow for review of each subject’s outcomes following the initial i.v. adminstration.  This staggered 
enrollment and dosing will be in-place for the first 6 subjects treated.  The staggering will be performed 
as follows: 
 
The second subject will be treated (dosed) only after the first subject has completed the Month 
1 safety visit and the Investigator and Medical Monitor have reviewed those data.   
 
Treatment (dosing) of subsequent subjects will be managed in a similar fashion, with subjects 
treated at a rate of one subject per month, with Investigator and Medical Monitor review of the 
Month 1 safety and tolerability outcomes for each subject before treatment of the subsequent 
subject is performed.   
This will continue through the 6th subject.  Assuming an acceptable safety and tolerability profile is 
observed, enrollment of the remaining subjects will be determined by the site’s accrual rate. 
Data Safety Monitoring Board (DSMB) 
A DSMB will be established to monitor subject safety. The DSMB will initially meet after 3 subjects 
and then after 6 subjects and then again after 15 subjects have received at least one dose of double-blind 
study medication and have completed their in-clinic visit through at least one month post-dose.  The 
DSMB will review unblinded data to assess the safety and tolerability of the treatments given.  The 
DSMB will, following each of these meetings, provide the Sponsor with a summary of their findings 
and recommendations (if any) for protocol mediation/amendment.   
The DSMB Charter will describe the full scope of DSMB activities, roles and responsibilities. 
Number of subjects (planned):  
Up to approximately 24 subjects will be randomized (in a 2:1 ratio, AVT001:Placebo), and thus up to 
approximately 16 subjects will be treated with AVT001 and 8 subjects will be treated with placebo. 
Diagnosis and main criteria for inclusion:  
Inclusion Criteria 
Subjects who meet all of the following criteria are eligible for enrollment as study participants: 
1. 
Diagnosis of type 1 diabetes, within 12 months of first dosing, confirmed by positive lab 
result for one or more of the following types of  autoantibodies: 
• 
Glutamic acid decarboxylase (GAD65) 
• 
Insulinoma associated protein 2 (IA-2, also known as ICA-512) 
• 
Zinc transporter 8 (ZnT8) 


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2. 
Age 16 or older and able to provide informed consent/assent. 
3. 
   Has a CD8+ T cell defect that is correctable in vitro by co-culture with immature dendritic 
cells loaded with a synthetic oligopeptide from Hsp60sp. 
4. 
If a participant is female with reproductive potential, willing to avoid pregnancy through the 
duration of the trial.  
5. 
Signed and dated written informed consent/assent. 
Exclusion Criteria 
Subjects who meet any of the following criteria are not eligible for enrollment as study participants: 
1. 
Poorly controlled diabetes despite insulin therapy, who in the opinion of the investigator 
would not be a good candidate for participation in a clinical trial. 
2. 
Screening hemoglobin <10.0 g/dL; leukocytes <3,000/uL; neutrophils <1,500/uL; 
lymphocytes <800/uL; platelets <100,000/uL 
3. 
Screening Urine Albumin Excretion > 300mg/gmCr 
4. 
Screening eGFR < 60 mL/min/1.73m
2
 
5. 
Screening ALT or AST > 1.5x upper limit of normal (ULN) 
6. 
Screening bilirubin > 2.0 mg / dL, or > 3.0 mg / dL for participants with Gilbert’s Syndrome 
7. 
Current use of immunosuppressive or immunomodulatory therapies, including 
pharmacologic doses of systemic steroids (topical steroidal creams and inhaled steroids 
without large systemic absorption are allowed). 
8. 
Coincident medical condition likely to require immunosuppressive or immunomodulatory 
therapies 
9. 
Coincident medical condition likely to limit short term (5 year) life expectancy (malignancy, 
symptomatic coronary artery disease, recent stroke) 
10. 
Prior radiation therapy, immunotherapy (within 1 year of screening), or chemotherapy 
11. 
Serologic evidence of current HIV-1or HIV-2 infection 
12. 
Serologic evidence of hepatitis C infection 
13. 
Serologic evidence of acute or chronic active hepatitis B as measured by Core Ab positive 
and / or Surface Antibody antigen positive  
14. 
Subjects with other autoimmune conditions (except compensated or treated autoimmune 
thyroid, celiac, alopecia, or vitiligo diseases) 
15. 
Women who are pregnant (pregnancy testing during screening), breastfeeding, or planning 
pregnancy during the study period. 
16. 
Inadequate venous access to support leukapheresis 
17. 
Any condition that in the opinion of the investigator would preclude the subject from 
participating in a clinical trial. 
18. 
Abnormal screening ECG that in the opinion of the investigator or sponsor would pose a 
safety risk 
Investigational product, dosage and mode of administration:  
Formulation of dosage: AVT001 will be cryopreserved in infusible cryomedia in cryopreservation 20 
mL bags. Each infusion bag will contain between 7 x 106 and 10 x 106 cells, and each subject 
randomized to AVT001 will have 3 such bags manufactured for infusion. The cryopreserved cells will 
be manufactured at the Dana Farber Cancer Institute Cell Manipulation Core Facility (DFCI-CMCF) 
and will be transported to the site for infusion. 


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Route of administration: The infusions will be administered intravenously. 
Frequency of administration: Three (3) infusions will be administered approximately 30 (+/- 7) days 
apart.  Infusions must be at least 21 days apart. 
Duration of treatment:  
There are three pre-defined periods in this study. 
 
Screening and cell collection period will last up to 3 months. 
 
Treatment period will consist of 3 doses, each ~30 days apart (Baseline, Month 1, Month 2) 
and a 1-month post-last dose assessment period (through Month 3).  Thus, the treatment period 
is defined to be 3 months in duration. 
 
Post-Treatment Follow-up Period will extend approximately 21 additional months (and thus 
through Month 24 of the study).  
Reference therapy, dosage and mode of administration:  
Matching placebo will have similiar intravenous infusion 
Criteria for evaluation: 
Safety (Primary Endpoint): 
Consistent with the study objectives, the primary endpoints will focus on the safety and tolerability of 
AVT001 versus placebo.   
The primary endpoints are: 
(a) The incidence of treatment-emergent adverse events (TEAEs).   
(b) Clinically significant changes from baseline in clinical laboratory parameters  
(c) The incidence and severity of local i.v.-site reactions 
(d) Changes from baseline in vital signs and electrocardiograms 
(e) Incidence of severe hypoglycemic events  
Efficacy (Secondary/Exploratory Endpoints): 
Secondary objectives focus on the CD8+ T-cell regulatory system and therapeutic outcomes associated 
with T1D.  These pharmacodynamic effects of AVT001 will be assessed in an exploratory manner over 
the course of the study period. 
Secondary endpoints will include: 
(a) Assessment of the HLA-E-restricted CD8+ T cell regulatory activity (“potency assay”) 
(b) Changes from baseline in the area under the curve (AUC) of the stimulated C-peptide levels 
over a 4-hour mixed meal tolerance test (MMTT) 
(c) Changes from baseline in HbA1c 
(d) Change from baseline in insulin usage 
(e) Changes from baseline in autoantibody levels  
Statistical methods: 
Sample Size 
The primary objective of this study is to assess the safety and tolerability of AVT001, as compared to 
placebo, in subjects with type 1 diabetes.  A sample size of 14 subjects treated with AVT001 will 
provide at least 80% power to observe at least one occurrence of treatment-emergent adverse events 
with a true underlying incidence of 12%.  The sample size provides the same power to detect at least 
one occurrence of local tolerability events relating to the i.v. administration of study drug.   


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Secondary objectives of this study include assessment of changes in pharmacodynamic markers, 
including increases of the function in the CD8+ T-cell regulatory system and improvements of C-
peptide levels, and HbA1c.  The sample size of 14 subjects treated with AVT001 and 7 subjects treated 
with placebo will provide estimates of the mean and standard deviation of the treatment benefit of 
AVT001 as compared to placebo in these endpoints.   
To account for an estimated 10% potential dropout prior to the completion of the Month 5 
safety/efficacy assessments, a total sample size of approximately 24 subjects will be randomized, in a 
2:1 ratio of AVT001:Placebo.   
Analysis Timing 
The primary analysis for this study will be performed after the last subject has completed the Month 5 
study visit, at which time the study database will be locked and the treatment allocation codes 
unblinded for analysis.  This analysis will form the primary basis for the assessment of the study 
objectives. 
Continuing data collection through the long-term follow-up will be performed through Month 24.  
While Investigators and subjects will remain blinded to the treatment allocations, assessment and 
analysis of outcomes will be performed by the Sponsor in an on-going manner, to better understand the 
longer-term effects (on both safety and efficacy) of the therapy. 
Safety Assessment 
Adverse events will be reported using the MedDRA coding dictionary.  Tabulations will include an 
overall incidence of at least one adverse event, incidence within body system, and incidence by 
preferred term.  Each subject may only contribute once (ie, first occurrence) to each of the incidence 
rates, regardless of the number of occurrences.  The incidence of adverse events will be presented as 
follows: 
 
The incidence of treatment-emergent adverse events (TEAEs), ie, those events that start on 
or after the first dose of study medication, will be tabulated by treatment for the Treatment 
Phase.  This will include events starting within 30 days following the last dose. 
 
The incidence of post-treatment events (those events that start more than 30 days following 
the last dose) will be tabulated. 
Subjects with serious adverse events (including deaths) and subjects who discontinue due to adverse 
events will be listed.  Subjects who have other significant serious adverse events deemed to be of 
special interest because of clinical importance will also be listed.   
Laboratory, vital signs, and ECG outcomes will be assessed for changes over time as well as clinically 
significant shifts from pre-treatment.  The incidence of severe hypoglycemic events will be tabulated. 
Efficacy Assessment 
Changes over time in C-peptide levels, HbA1C, total daily insulin dose and antibody values will be 
assessed.  For purposes of data presentation, a mixed model for repeated measures (MMRM) will be 
used to present changes in these parameters over time.  Details regarding this MMRM will be provided 
in the SAP.  The SAP will also detail the HLA-E restricted CD8+ regulatory T cell function analysis.


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4 
Introduction 
This document is a protocol for a human research study to be conducted according to US and 
international standards of Good Clinical Practice (FDA Title 21 part 312 and International  
Conference on Harmonization guidelines), applicable government regulations and Institutional 
research policies and procedures. 
4.1 
Background 
Type 1 diabetes (T1D) is one of the most common chronic diseases of childhood. It accounts for 
approximately two-thirds of all cases of diabetes in subjects ≤19 years of age. According to 
statistics reported by the Juvenile Diabetes Research Foundation (JDRF), it is estimated that as 
many as 3.0 million subjects have T1DM in the United States, with more than 15,000 children and 
15,000 adults diagnosed each year.  
T1D is a complex, chronic, T cell-mediated autoimmune disease in which the insulin producing 
pancreatic β cells are destroyed. In T1D, there is a loss of tolerance to β cells that results in a CD4 
and CD8 T cell- dependent autoimmune process that culminates in complete destruction of insulin-
producing β cells, leading to insulin deficiency and dysregulated glucose metabolism. Diabetic 
subjects manage their hyperglycemia by daily insulin injections, however this treatment does not 
cure the disease or prevent the possibility of the disease’s serious effects, which may include kidney 
failure, blindness, nerve damage, heart attack, stroke, and pregnancy complications (JDRF website). 
It is important to develop other therapies able to target the primary defect of the immune function 
that causes T1D. As discussed below, immunotherapeutic approaches that could potentially treat and 
or prevent T1D have been moving into the clinic recently. Over the years, studies using the non-
obese diabetic (NOD) mouse model have provided important insights into the immunology of T1D 
and have demonstrated the feasibility of preventing or curing T1D in an antigen-specific manner by 
specifically manipulating disease-relevant autoreactive T cell populations without compromising the 
immune system at large. 
Conceptually speaking, selective control of unwanted immune responses could be achieved by 
induction of “antigen-specific tolerance”, provided that the specific antigens that elicit the unwanted 
immune responses have been identified. However, for decades, it has always been a tremendous 
challenge to identify the exact pathogenic peptide/s for any given human organ specific 
autoimmune diseases, or specific allo-antigen peptide/s responsible for graft rejection. In reality, no 
definitive  answers have ever been obtained for any human autoimmune diseases yet. Furthermore, 
induction of “antigen specific tolerance” requires precise knowledge of the peptide associated with 
particular MHC molecules and highly polymorphic human HLA system inevitably increases the 
uncertainty of searching for such MHC/peptide complex in each subject. Thus, the necessity of 
identification of a precise MHC/peptide complex for each individual from an unknown and 
countless antigen  specificity  pool makes the induction of “antigen-specific tolerance” an 
unfeasible approach to specifically and effectively treat organ specific autoimmune diseases and/or 
control graft rejection in clinical immunology. In contrast, Qa-1/HLA-E restricted CD8+ T cells, 
provide a simple and unified “class action” of self-nonself discrimination, regulating the unwanted 
immune responses, independent of the knowledge of  the particular pathogenic peptides in 
autoimmune diseases and/or graft rejection for each subject, currently largely undetermined. (1, 2). 
Avotres has developed a novel approach to manipulate autoreactive T-cell responses against all self 
antigens, including -cell antigens (1, 2), that we would  like to evaluate in this clinical protocol. 
Modulating the activity of self-reactive T cells to damper autoimmunity at a system level without 
damaging normal on-going anti-infection and anti-tumor immunity represents an ideal new 


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approach to treat T1D. 
4.2 
Failure of self-nonself discrimination by the immune system results in pathogenesis of 
autotimmune disease such as T1D 
The pioneering work of Burnet and Medawar clearly indicated that self-nonself discrimination is 
not achieved by recognizing the structural differences between self antigens and foreign antigens, 
because introducing a foreign allo transplantation antigen into the immune system early enough in 
life renders the recipient animals tolerant to that foreign antigen which is subsequently treated as if 
it were a self antigen (3, 4). It is well known now that thymic negative selection that deletes high 
avidity T cells  specific to any antigen, self-antigens as natural cases, is the major mechanism of the 
specific tolerance established (5-8). However, as a biological consequence of natural thymic 
negative selection, self-reactive T cells with intermediate and low avidity escape into the periphery 
(9-11). The intermediate avidity self-reactive T cell pool in the periphery represent a potential 
danger of pathogenic  autoimmunity (1, 12), which could be accidentally activated by a sufficiently 
high level of self-antigens to initiate organ-specific autoimmune diseases during infections or 
injuries (1, 11, 13-15) and must be effectively controlled. We have thus postulated that self-nonself 
discrimination must continue in the periphery after the thymic negative selection to maintain self-
tolerance (1, 12). In this regard, it has been shown that Qa-1/HLA-E restricted CD8+ T cell 
mediated regulatory pathway is important in maintaining peripheral selftolerance (16-24). We have 
further proposed and tested an “Avidity Model Of Peripheral T Cell Regulation”, in which Qa-
1/HLA-E restricted CD8+ T cells, by selectively down-regulating the intermediate avidity T cell 
pool, play a central role in control of organ specific autoimmune diseases in the conventional 
soluble protein antigen system in both mice (19, 20, 22) and humans (23). The peripheral self-
reactive T cell repertoire is devoid of high avidity T cells due to thymic negative selection. 
Therefore, selective down-regulation of the intermediate avidity T cell pool enables the selective 
suppression on the overall immune responses to self-antigens, mediated by the intermediate avidity 
T cells, but not on effective immune responses to foreign antigens dominated by the high avidity T 
cells. This way, the Qa-1/HLA-E restricted CD8+ T cells are able to specifically control organ-
specific autoimmune diseases or establish donor specific transplantation tolerance without 
damaging ongoing effective anti-infection and anti-tumor immunity (2, 19, 20, 22, 23, 25). The 
selective down-regulation of the intermediate avidity T cell pool by the Qa-1/HLA-E restricted 
CD8+ T cells is accomplished by a specific recognition of the unique common target structure, the 
signal peptide of heat shock protein 60 (Hsp60sp) associated with the non-polymorphic MHC Class 
1b molecule Qa- 1/HLA-E, preferentially expressed on the intermediate avidity T cells (19, 20, 22, 
23). 
More relevant, we have shown a defect in the T cell regulatory pathway  which  normally controls 
autoreactive T cells that attack the body’s own tissues and organs (23). A majority of people with 
Type 1 diabetes who were tested in the study were found to have this newly-identified 
cellular/molecular defect on HLA-E restricted CD8+ T cells, and we were able to successfully 
correct the defect in-vitro in most of the subjects tested by retriggering the CD8+ T cells with 
Avotres in vitro made of the autologous DC loaded with the synthetic oligopeptide from Hsp60sp 
(23). 
Our preclinical human in vitro studies and murine in vivo studies formed the scientific basis for this 
IND application for trying AVT001 in vivo. 
4.3 
The Autoimmune Process of Type 1 diabetes 


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Development of autoimmune disorders is a result of breakdown in mechanisms that maintain 
unresponsiveness to self. Specifically, T1D is an automimmune disorder in which T cells are self-
reactive or activated T cells that are activated against major antigenic components of pancreatic β 
cells are out of the control by the normal regulatory mechanisms. While these self-reactive T cells 
are under the control of peripheral regulatory mechanisms in healthy individuals, failure of control 
leads to the destruction of the β cells and consequent T1D. The autoimmune attack on pancreatic β 
cells is orchestrated by a  variety of cells that produce cytokines and other toxic mediators. These 
self-reactive T cells work together with other lymphocytes and antigen-presenting cells to mediate 
this damage and have been shown in animal models to be important both in the early stages of 
diabetes development and in the final effector stages. They kill self-reactive T cells specific to self-
antigens, which include  cell antigens, insulin and other key proteins associated with beta cell 
functions. 
4.4 
Autoreactive T cells in T1D 
As reviewed in Wong 2008, many studies carried out in the NOD mouse model of T1D have 
indicated that both CD4+ and CD8+ T-cells inflict islet β-cell damage both at an early stage in 
diabetes development and at the final effector phase (26-28). There appears to be considerable 
similarity between the antigens recognized in humans and in the NOD animal models. In human 
studies, so far limited to testing peripheral blood lymphocytes, a variety of reactivities to putative 
autoantigens have been found, including proinsulin, GAD65, ppIAPP (pre-pro-islet amyloid 
polypeptide protein) and GFAP (glial fibrillary acidic protein). By the time diabetes occurs, it 
appears that there may be a broad spread of reactivity, which is different for each subject, and it will 
continue to be a major challenge to identify those antigenic epitopes that are important in the early 
phases of pathogenesis. Many of these antigenic epitopes represents attractive targets to go after for 
T1D therapeutic interventions. 
Mechanisms by which T cells cause damage include production of cytotoxic cytokines such as IFNγ 
(interferon γ) and TNFα (tumour necrosis factor α), that have a direct cytotoxic action on the islets. 
In addition, these cytokines have an important role in up-regulation of MHC class I molecules, 
which could increase immune recognition of the cells. These T-cell-generated inflammatory 
cytokines can work together with cytokines produced by macrophages, such as IL (interleukin)-1, to 
have a synergistic damaging effect on islet β-cells. The CD8+ CTLs also exocytose granules from 
secretory lysosomes which contain cytotoxic proteins that include the pore-forming protein, 
perforin, and serine esterases, called granzymes. In summary, autoreactive T cells are considered as 
emerging targets for prospective antigen- specific therapeutic interventions. 
4.5 
Immunotherapeutic approaches to treat T1D 
Many broad based immunosuppressive and antigen-specific immunoregulatory therapies have been 
and are currently being evaluated for their utility in the prevention and treatment of T1D. These 
may be administered as monotherapeis or in combination. Some of the below-mentioned have 
shown efficacy in initial clinical trials. Many tend to be small molecule inhibitors and monoclonal 
antibodies directed against cytokine and cytokine receptors, proteases, CD3 or CD20 receptors (i.e. 
Rituximab), GAD65, CTL14, heat shock proteins, or insulin-related molecules (29). Systemic non-
antigen specific immunomodulator interventions include cycyclosporine A, azatiopirine, and anti-
thymocyte globulin (ATG) plus prednisolone. 
Other cell therapies using dendritic cells are also currently being tested as described briefly below. 
Additional examples of cell therapies include targeting regulatory T cells (Tregs). Approaches to 


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isolate and expand polyclonal and antigen-specific Tregs in vitro have been developed for 
immunotherapy and a clinical trial to test this hypothesis in the US (47) 
Unlike those small molecule therapies and any currently tried cell-based therapies, AVT001 
provides for a potential new type of cell-based therapeutic approach to treat and/or prevent T1D. 
AVT001, used as a vaccine, it may activate and correct the defective regulatory pathway mediated 
by HLA-E restricted CD8+ T cells in the identified subjects that normally function to selectively 
control the pathogenic self-reactive T cells that destroy β cells without damaging the normal on 
going anti-infection and anti-tumor immunity, a process called self-nonself discrimination (19, 20, 
22, 23). 
4.6 
Dendritic cell based therapies for T1D 
Dendritic cells (DCs) were first described by Ralph Steinman (30) nearly forty years ago.  It is 
known that  DCs existed in all lymphoid and most non-lymphoid tissues. DCs are antigen-
presenting cells (APCs),  which play a critical role in the regulation of the adaptive immune 
response. To perform this function,  DCs are capable of capturing antigens, processing them, and 
presenting them on the cell surface along with appropriate costimulation molecules. The crucial 
function of DCs falls broadly into two categories, each of which involves antigen presentation. The 
first category of DCs function is antigen presentation and activation of T cells. The second category 
of DCs function has been suggested that a different class of DCs exist with the function of inducing 
and maintaining immune tolerance. 
Cell therapy using dendritic cells use “immature” dendritic cells that are in vitro derived from 
monocyte precursors isolated from diabetic subjects and then either modified with and reinjected 
into the same individuals with the hope of resetting the immune system balance in the individual. 
There is a great deal of interest in how DCs might be exploited as a form of immunotherapy. DCs 
are being studied as adjuvants for vaccines or as a direct therapy to induce immunity against 
cancer. Human clinical trials are ongoing to use DCs to induce immunity to antigens against 
prostate cancer, lung cancer, melanoma and multiple myeloma etc. The first attempt to use DCs as 
cancer vaccines in humans was made by Edgar Engleman and Ronald Levy (31), who isolated DCs 
from subjects with non-Hodgkin's lymphoma who had failed conventional chemotherapy, loaded 
the cells with immunoglobulin idiotype obtained from the subject's tumor, and re-injected the 
antigen-loaded cells back into the subjects. Four subjects involved in this pilot clinical trial and all 
treatments were well tolerated, no significant side effects were associated with the infusion of 
antigen-loaded DCs. Since then, pilot clinical trials of antigen-pulsed DCs have been conducted in 
various types of cancer, including melanoma, prostate cancer, multiple myeloma, and non- small 
cell lung cancer. DC-based therapy as immunotherapy is also tried in human type 1 diabetes and a 
clinical trial to test this hypothesis has finished its phase I study in the US (Clinical Trials. gov 
identifier NCT00445913) (32). In general, it is widely accepted that DC-based therapy is consider a 
very safe therapy. 
4.7 
Dosing Rationale 
Our preclinical human in vitro studies and murine in vivo studies formed the scientific basis for this 
clinical application to assess AVT001 in vivo. AVT001 is the study product name for autologous 
immature dendritic cells loaded with the synthetic oligopeptide from Hsp60sp.  The study product 
represents an individualized preparation of autologous immature dendritic cells, derived by culture 
of the subject’s adherent primary monocytes with GM-CSF and IL-4 for 6 days, and loaded 
passively with the synthetic oligopeptide from Hsp60sp (QMRPVSRVL). In this study design 


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described in this protocol, three vaccines will be administered every 30 days (+/- 7 days but at least 
21 days apart). 
We feel strongly that the selection of the proposed route of administration, dose levels, dosing 
regimen, timing of vaccination relative to disease state, etc. in our clinical trial should rely on the 
information from similar human clinical trials rather than from animal studies. Particularly, it 
should not be heavily dependent on NOD/T1D model, which is a spontaneous disease animal model 
involving multiple mechanisms other than Qa-1 restricted CD8+ T cells and may not be exactly the 
same as human T1D subjects that we plan to test. For example, the subjects we propose to test are 
clearly defined to be the new onset T1D subjects who have the defect of the HLA-E restricted 
CD8+ T cells and the defect can be corrected in vitro by retriggering with AVT001. This design is 
based on the studies from human T1D subjects and its clinical regimen cannot be exactly mimicked 
and represented by the studies from NOD/T1D model. The best example in this case, which is 
almost identical to our circumstances is an iDC based therapy on T1D subjects (48) (32) versus its 
preclinical studies on NOD/T1D model (33), in that injections of 10 x 106 iDC every 2 weeks for 
total four times in humans versus a single injection of 2-3× 106 iDC in NOD mice. 
In NOD mice we have used 0.5-2 x 106/mouse without any abnormality in protected mice, which 
dose range would be proportionally about 50-100 x 106/person in humans. However, in search for 
the information in other similar human clinical trials, the dosage range used has been reported to be 
between 2-50 x 106/dose every 2 weeks from two to 13 doses and showed no significant side effects 
(see Table below). Particularly, in one recently completed trial using iDC onT1D subjects, 10 x 106 
iDC every 2 weeks for total four times were used (32), which is very close to our proposal, eg, 10 x 
106 iDC every 4 weeks for total three times will be used. Finally, from our own studies, HLA-E 
restricted CD8+ T cells represent a very potent subset of CD8+ T cells, probably account for 1-2% 
of total CD8+ T cells. We calculate if the iDCs needed for activate such CD8+ T cells is at a ratio of 
1:50-100, 10 x 106 of iDC would activate 500-10,000 x 106 of HLA-E restricted CD8+ T cells, 
which is in the range of 1-2% of total 10 x 1012 CD8+ T cells. 
Table 1:  Summary of dosage and route of administration of dendritic cell-base vaccines in 
completed human clinical trails, which have been concluded as safe therapies 
Therapy to 
Dose/schedule 
Route 
Number 
Subjects
Ref 
NHL 
(Non-Hodgkin’s 
lymphoma) 
2-17×10
6 / 4 weeks × 4 
i.v. 
35 
(34) 
 
2-17×10
6 / 4 weeks × 4 
i.v. 
10 
(31) 
Melanoma 
5-50×10
6 / 2 weeks × 4 
i.v./i.d./i.n.
28 
(35) 
 
5-50×10
6 / 2 weeks × 4 
i.v. 
14 
(36) 
 
3×10
6+6/12×10
6 / 2 weeks × 
5 
i.d.+i.v. 
13 
(37) 
Myeloma 
0.5-11×10
6 / 4 weeks × 2 
i.v. 
12 
(38) 
 
3.5-89×10
6 / 2 weeks × 3 
i.v. 
6 
(39) 
Prostate cancer 
1-20×10
6 / 6-8 weeks × 4-5 
i.v. 
51 
(40) 


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10×10
6/ 2 weeks × 4 
i.v.+i.d. 
8 
(41) 
 
0.3-40×10
6 / 4 weeks × 2 
i.v./i.d./i.l.
21 
(42) 
 
10×10
6/ 2 weeks × 3 
i.v.+i.d. 
13 
(43) 
 
0.94-2.02×10
8/ 4 weeks × 3 
i.v. 
14 
(44) 
RCC 
(Renal cell 
carcinoma) 
5-10×10
6 / 4 weeks × 3-13 
i.v./i.d. 
35 
(45) 
 
10-50×10
6 +10×10
6  /2 
weeks × 3 
i.v.+i.d. 
10 
(46) 
Type 1 diabetes 
10×10
6 / 2 weeks × 4 
i.d. 
10 
(32) 
 
Based on the information from our own studies and the published results from clinical trials on DC-
based therapies we have chosen a relatively low dose and regimen of AVT001 for safety 
consideration with a maximum potential for efficacy in vivo in our proposal. 
We choose to vaccinate newly diagnosed T1D subjects that will be determined to have the defect 
of HLA-E restricted CD8+ T cells and the defect could be corrected by co-culture with AVT001 
in vitro.  
According to a 2017–2018 National Health Interview Survey (NHIS), National Center for Health 
Statistics, and Centers for Disease Control and Prevention report, as of 2018 an estimated 187,000 
children and adolescents younger under 20 years of age had been diagnosed with with type 1 diabetes 
(49).  Following the DSMB review of the safety data for the first 6 staggered adult subjects (18 years 
and older), no safety concerns were identified.  Therefore, due to the safety established thus far and 
due to the high prevalence of disease in adolescents, this protocol is being amended (Amendment # 
3) in a step-wise fashion to reduce the lower limit of age for eligibility to age 16.   
 
 


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5 
Study Objectives and Design 
Our working model for maintenance of T cell self-tolerance holds that a subset of HLA-E restricted 
regulatory CD8+ T cells that specifically recognize the target structure of Hsp60sp associated with 
HLA-E expressed on the pathogenic self-reactive T cells and selectively down-regulate these 
pathogenic T cells which are responsible for autoimmune diseases (19, 20, 22, 23). The failure of 
this HLA-E-restricted  CD8+ T cell subset to recognize, or to suppress, its target population results 
in a permissive state in which organ-specific autoimmunity can emerge. We have identified defects 
in this CD8+ T cell system among nine of ten T1D subjects evaluated, and found the defect to be 
reversible in most of these in vitro by co-incubation with the synthetic oligopeptide from Hsp60sp-
loaded autologous immature dendritic cells (AVT001). 
5.1 
Primary Outcomes 
 To determine the safety and tolerability profile of three monthly doses of AVT001 
(as compared to placebo) in subjects with T1D. 
5.2 
Secondary Outcomes 
 To determine whether, among T1D subjects with in vitro-reversible defects in the 
CD8+ T cell regulatory system, infusion of AVT001 will lead to an in vivo 
reconstitution of this system. 
 To determine whether, among T1D subjects with in vitro-reversible defects in the 
CD8+ T cell regulatory system, infusion of AVT001 will result in amelioration of  
cell destruction as defined by preservation of stimulated C-peptide. 
5.3 
General Study Design 
This is a double-blind, randomized, placebo-controlled study to evaluate the safety and tolerability of 
AVT001 cell-based therapy, and to assess its potential efficacy for the treatment of type 1 diabetes 
(T1D).  A brief overview of the trial design is outlined in a schematic form in Figure 1. 
. 
 


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Figure 1:  Study Design Schematic (Protocol AVT001-T1D-01)1 
 
 
 
The trial will involve approximately 24 new-onset T1D subjects who have been identified as having: 
(a) a defect in HLA-E-restricted CD8+ T cell function putatively associated with pancreatic β cell 
destruction and (b) evidence that this defect of HLA-E-restricted CD8+ T cells can be corrected by 
in vitro exposure to AVT001.  
Participants will be randomized, in a 2:1 ratio (AVT001:Placebo) to receive AVT001 or placebo by 
intravenous infusion.  
 AVT001: 16 subjects to receive AVT001 through i.v. administration.   
 Placebo:  8 subjects to receive saline through i.v. administration.   
Subjects will be treated with 3 consecutive doses of AVT001, administered with one month (+/-7 
days) intervals between doses.  The primary time point for assessment will be 3 months post-last 
dose (Month 5), with longer-term follow-up through Month 24. 
Administration of AVT001 will be performed in a double-blinded fashion as follows: the dosage 
regimen will be between 7 x 106 and 10 x 106 cells cells/infusion for each of the three infusions 30 
(+/- 7) days apart.  Each infusion will be administered by qualified clinical personnel in an 
established clinical research facility.  
Subjects will be monitored for physical or reported symptoms as well as for vital signs (including 
temperature, heart rate, blood pressure, and arterial oxygen saturation) throughout the infusion 
procedure and for a minimum of 2 hours following the procedure. Although unlikely to be required, 
the clinical facility will include at the subject’s side all medications and equipment necessary to 
treat cytokine release syndrome, acute anaphylactoid/infusion reactions, respiratory distress, and 
cardiac arrhythmia. 
A placebo control arm using saline and DMSO is being used as the comparator.  
 
1 Amendment # 3 includes participants aged 16 and older.  


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5.4 
Study Duration 
There are three pre-defined periods in this study. 
 Screening and cell collection period will last up to 3 months 
 Treatment period will consist of 3 doses, ~30 days apart (Baseline, Month 1, Month 2) and a 
1-month post-last dose assessment period (through Month 3).  Thus, the treatment period is 
defined to be 3 months in duration. 
 Post-Treatment Follow-up Period will extend 21 additional months (and thus through Month 
24 of the study). 
5.5 
Subject Enrollment 
The justification for the study design derives from the primary outcome of the proposed trial to 
assess the safety and tolerability of this therapy.  This is a first time in man study and therefore 
exposure will be managed using a staggered enrollment that will allow for review of each subjects 
outcomes following the initial IV.  This staggered enrollment and dosing for the first 6 subjects 
treated will be performed as follows: 
 The second subject will be treated (dosed) only after the first subject has completed through 
the Month 1 safety visit and the Investigator and Medical Monitor have reviewed those data.   
 Treatment (dosing) of subsequent subjects will be managed in a similar fashion, with 
subjects treated at a rate of one subject per month, with Investigator and Medical Monitor 
review of the Month 1 safety and tolerability outcomes for each subject before treatment of 
the subsequent subject is performed.   
This will continue through the 6th subject.  Assuming an acceptable safety and tolerability profile is 
observed, enrollment of the remaining subjects will be determined by the sites accrual rate. 
5.6 
Data Safety Monitoring Board (DSMB) 
A DSMB will be established that will monitor subject safety as well as data quality and adherence 
to safety rules. Additionally, the DSMB will review all safety/toxicity data for the trial and 
recommend trial modifications, suspension or termination, as needed.  
The DSMB will meet after 3, 6 and 15 subjects have received at least one dose of double-blind 
study medication and have completed their in-clinic visit through at least one month post-dose.  The 
DSMB will review unblinded data to assess the safety and tolerability of the treatments given.  The 
DSMB will, following each of these meetings, provide the Sponsor with a summary of their 
findings and recommendations (if any) for protocol amendments.   
A DSMB Charter will be written, to include details regarding meeting frequency, format, and 
membership. 
5.6.1 Stopping Rules 
Subjects will be clinically evaluated for evidence of toxicity from ≥ grade 3 toxicity as determined by 
CTCAE which is unmanageable, unexpected and (possibly, probably or definitely) related, and 
attributable to protocol therapy.  Termination  of enrollment would be considered with 2 dose 
limiting toxicities (DLTs) in the first 6 evaluable subjects and 3 in the first 9 evaluable subjects.  In 
the event of a DLT for a given subject, doses will be discontinued, the subject clinically evaluated, 
and taken off study. The study will be stopped for DLT resulting in death of a subject. 


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5.7 
Study Endpoints 
Consistent with the study objectives, the primary endpoints will focus on the safety and tolerability 
of AVT001 treatment versus placebo.   
The primary endpoints are: 
(a) The incidence of treatment-emergent adverse events (TEAEs).   
(b) Changes from baseline in clinical laboratory parameters.  
(c) The incidence and severity of local IV-site reactions.  
(d) Changes from baseline in vital signs and electrocardiograms.  
(e) Incidence of severe hypoglycemic events.  
Secondary objectives focus on the CD8+ T-cell regulatory system and therapeutic outcomes 
associated with T1D.  These pharmacodynamic effects of AVT001 will be assessed in an exploratory 
manner. 
Secondary endpoints will include: 
(a) Assessment of the HLA-E-restricted CD8+ T cell regulatory activity (“potency assay”) 
(b) Changes from baseline in the area under the curve (AUC) of the stimulated C-peptide levels 
over the first 4 hours of a mixed meal tolerance test (MMTT) 
(c) Changes from baseline in HbA1c 
(d) Change from baseline in total daily insulin dose 
(e) Changes from baseline in autoantibody levels 
The primary time point for analysis is Month 5, at which point the primary assessment of the study 
objectives is performed. 


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6 
Subject Selection and Withdrawal 
6.1 
Inclusion Criteria 
Subjects who meet all of the following criteria are eligible for enrollment as study participants: 
1. Diagnosis of type 1 diabetes, within 12 months of first dosing, confirmed by positive lab 
result for one or more of the following types of autoantibodies: 
a. Glutamic acid decarboxylase (GAD65) 
b. Insulinoma associated protein 2 (IA-2, also known as ICA-512) 
c. Zinc transporter 8 (ZnT8). 
2. Age 16 or older and able to provide informed consent/assent. 
3. Has a CD8+ T cell defect that is correctable in vitro by co-culture with immature dendritic 
cells loaded with the synthetic oligopeptide from Hsp60sp. 
4. If a participant is female with reproductive potential, willing to avoid pregnancy through the 
duration of the trial.  
5. Signed and dated written informed consent/assent. 
6.2 
Exclusion Criteria 
Subjects who meet any of the following criteria are not eligible for enrollment as study participants: 
1. 
Poorly controlled diabetes despite insulin therapy, who in the opinion of the investigator 
would not be a good candidate for participation in a clinical trial 
2. 
Screening hemoglobin <10.0 g/dL; leukocytes <3,000/uL; neutrophils <1,500/uL; 
lymphocytes <800/uL; platelets <100,000/uL 
3. 
Screening Urine Albumin Excretion > 300mg/gmCr 
4. 
Screening eGFR < 60 mL/min/1.73m
2 
5. 
Screening ALT or AST > 1.5x upper limit of normal (ULN) 
6. 
Screening bilirubin > 2.0 mg / dL, or > 3.0 mg / dL for participants with Gilbert’s 
Syndrome 
7. 
Current use of immunosuppressive or immunomodulatory therapies, including 
pharmacologic doses of systemic steroids (topical steroidal creams and inhaled steroids 
without large systemic absorption are allowed). 
8. 
Coincident medical condition likely to require immunosuppressive or immunomodulatory 
therapies. 
9. 
Coincident medical condition likely to limit short term (5 year) life expectancy 
(malignancy, symptomatic coronary artery disease, recent stroke) 
10. 
Prior radiation therapy, immunotherapy (within 1 year of screening), or chemotherapy 
11. 
Serologic evidence of current HIV-1 or HIV-2 infection 
12. 
Serologic evidence of hepatitis C infection 
13. 
Serologic evidence of acute or chronic active hepatitis B as measured by Core Ab positive 
and / or Surface Antibody antigen positive  
14. 
Subjects with other autoimmune conditions (except compensated or treated autoimmune 
thyroid, celiac, alopecia, or vitiligo diseases) 


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15. 
Women who are pregnant (pregnancy testing during screening), breastfeeding, or planning 
pregnancy during the study period. 
16. 
Inadequate venous access to support leukapheresis. 
17. 
Any condition that in the opinion of the investigator would preclude the subject from 
participating in a clinical trial. 
18. 
Abnormal screening ECG that in the opinion of the investigator or sponsor would pose a 
safety risk. 
6.3 
Early Withdrawal of Subjects 
If a subject is discontinued from the study prematurely, the Investigator must select the primary 
reason for discontinuation on the End of Study eCRF.  In addition, every effort should be made to 
complete the assessments listed under the  “Early Term” column on the Schedule of Events.  
Subjects withdrawn from the study will be considered evaluable for statistical assessment. 
A subject may be removed from the study for the following medical or administrative reasons: 
 Adverse Event: If a subject experiences an adverse event that the subject finds 
unacceptable or that, in the judgment of the Investigator or the Medical Monitor presents 
an unacceptable consequence or risk to the subject, the subject may be discontinued from 
further participation in the study. 
 Administrative Discontinuation:  After consultation with the Investigator or Medical 
Monitor, a subject may be discontinued from the study for failure to comply with protocol 
requirements.  All instances of noncompliance must be documented in the eCRF. 
 Refusal of Assessments:  If for any reason, following dosing, the subject refuses further 
assessment during the study, the subject shall be discontinued from the study and the 
reasons for refusal documented.  Reasonable efforts shall be made to monitor the subject 
for adverse events following such discontinuation.  Such efforts shall be documented. 
Standard supportive therapy will be maintained for subjects withdrawn from active treatment.   Every 
effort will be made to collect safety data from these subjects through the end of study.   


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7 
Description of Study Drug 
A study drug handling manual and study drug administration manual (Manual of Operating 
Procedures, ie, MOP)  describes study drug management in detail.  The MOP includes procedures for 
collection, transport, processing, storage, preparation (for dosing), and administration of study 
medication, as well as details regarding how the blinding is to be maintained during the infusion 
process.  The following sections provide an overview of the key elements of those activities, but the 
MOP should be referred to for details. 
Drug name and active ingredients: The study drug represents an individualized preparation of 
autologous immature dendritic cells, derived by culture from the subject’s adherent primary 
monocytes with GM-CSF and IL-4 for 6 days, and loaded passively with the synthetic oligopeptide 
from Hsp60sp. 
Formulation of dosage: AVT001  will be cryopreserved in infusible cryomedia in infusion bags. 
Each infusion bag will contain between 7 x 106 and 10 x 106 cells, and each subject randomized to 
AVT001 will have 3 such bags manufactured for infusion. The cryopreserved cells will be 
manufactured at the Dana Farber Cancer Institute Cell Manipulation Core Facility (DFCI-CMCF) 
and will be transported to the investigational site for infusion. 
Route of administration: The infusions will be administered intravenously. 
Frequency of administration: Three (3) infusions will be administered 30 (+/- 7) days apart.  While 
there is a 7-day window for the infusions, they should occur at least 21 days apart. 
Table 2 provides a summary description of the drug products, including the dosage form, the unit 
dose, and a physical description of the products.  
Table 2: 
Investigational Product (Protocol AVT001-T1D-01) 
 
Investigational Product 
Product Name: 
AVT001 
Placebo 
Dosage Form: 
20 mL AVT001 
suspension with 
10% DMSO in 
Human Serum 
Albumin 
20 mL saline with 
10% DMSO in 
Human Serum 
Albumin 
Packaging: 
Cryopreservation 
bags 
Cryopreservation 
bags 
Route of Administration:
Intravenous (i.v.) 
Intravenous (i.v.) 
Physical Description: 
Blinded (covered) 
bottle/tube 
Blinded (covered) 
bottle/tube 
Manufacturer: 
 DFCI-CMCF 
 DFCI-CMCF 
7.1 
Administration of Study Drug. 


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A study drug handling manual and study drug administration manual (Manual of Operating 
Procedures, ie, MOP) describes study drug management in detail.   
Administration of the treatment regimen is as follows: 
 On Study Day 1, the subject will receive study treatment (the AVT001 or placebo  infusion). 
 The interval between injections will be 30 (+/- 7) days.  Subsequent injections will be at Study 
Day 30 and 60 (each ± 7 days and at least 21 days apart), for a total of 3 i.v. injections. 
Subjects will be observed for at least 2 hours post-end of each infusion.  Safety and tolerability 
assessments will be performed, including measurement of vital signs.  Local tolerability at the 
infusion site (such as pain, swelling, tenderness, and erythema) will be assessed via adverse event 
reporting. Please refer to the study drug handling manual and study drug administration manual 
(Manual of Operating Procedures, ie, MOP) for details on monitoring.Receiving, Storage, 
Dispensing and Return 
A study drug handling manual and study drug administration manual (Manual of Operating 
Procedures, ie, MOP) describes study drug management in detail. 
7.2 
Reconciliation of Drug  Product 
A study drug handling manual and study drug administration manual (Manual of Operating 
Procedures, ie, MOP) describes study drug management in detail. 


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8 
Study Procedures 
Study procedures are listed below and can also be found in the Schedule of Events (Table 3). No 
study procedures will be performed prior to obtaining informed consent. 
8.1 
Informed Consent 
The consent process will be conducted by qualified study personnel.  All participants (or their legally 
acceptable representative) must read, sign, and date a consent form prior to participation in the study 
and/or undergoing any study-specific procedures. 
The informed consent form must be updated or revised whenever important new safety information 
is available, when indicated for a protocol amendment, and/or whenever any new information 
becomes available that may affect a subject’s participation in the study. 
8.2 
Screening/Randomization/Leukapheresis  
8.2.1 Screening Study Day ≤ - 90  
Screening assessments may be performed at more than one visit, if needed, and must be completed 
prior to randomization, to establish subject eligibility. Unless otherwise noted, all tests will be 
required within 90 days prior to treatment. 
 Medical history including prior treatment, current medicines, and drug allergies 
 Physical exam including blood pressure, pulse, temperature, weight 
 Screening Laboratory studies including:  
o Complete Metabolic Panel (CMP) to include: glucose, BUN, creatinine, 
electrolytes, calcium, total protein, albumin, total bilirubin, AST, ALT, alkaline 
phosphatase 
o Complete Blood Count with differential (CBC w diff) [CBC may need to be 
repeated if not performed within acceptable safety timeframe prior to 
leukapheresis; see MOP] 
o HbA1c 
o eGFR (calculated) 
o Urine MicroAlbumin panel (urine microalbumin, urine creatinine, 
microalbumin/creatinine ratio) 
o Creatine Clearance (CrCl) (calculated) 
 Measurement of auto-antibodies, including: 
o Insulin associated aAb (IAA) 
o Insulinoma associated protein 2 (IA-2, also known as ICA-512) 
o Glutamic acid decarboxylast (GAD65) 
o Zinc transporter 8 (ZnT8). 
 Evaluation of HLA-E-restricted CD8+ T cells (“potency assay”). 
 A urine pregnancy test will be collected for females of child-bearing potential. 
 ECG 
 Viral studies comprising HIV-1, HIV-2, HBV Core Ab positive and / or Surface 
Antibody antigen positive, and HCV serology. 


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 Vein check 
Screening Lab eligibility requirements  
 Potency Assay results from Screening do not expire. 
 Auto-antibody and Viral testing results from Screening expire 6 months from collection. 
 All other safety labs results from Screening expire after 90 days from collection 
including urine pregnancy tests for women of child bearing potential. 
8.2.2 Randomization 
Upon successful completion of all eligibility requirements, study participants may be randomized to 
the trial.  Each subject will be randomized to treatment in a 2:1 ratio of AVT001:Placebo.  As this is 
a first-in-man study, block sizes of N=3 for the first six subjects will be used to allow for review of 
the initial six subjects with four AVT001 and two placebo-treated.  Thereafter, the block sizes will be 
randomly assigned to ensure the integrity of the randomization schema while maintaining a 2:1 ratio 
of AVT001 to placebo. 
8.2.3 Leukapheresis  
Following randomization, all subjects (including those randomized to placebo arm as well as the 
AVT001 arm) will undergo leukapheresis according to standard clinical procedures.   
The apheresis product will be delivered to Dana Farber Center Cancer Institute (DFCI), Cell 
Manipulation Core Facility (CMCF) for processing to produce AVT001.  Detailed scheduling, 
coordination, communication, handling, and other procedures for all vein-to-vein operations are 
described in the MOP. 
Leukapheresis is to be performed after randomization, with sufficient time for AVT001/placebo 
manufacture prior to the end of window between Screening Visit (Visit 1) and first infusion (Study 
Day 1, Visit 3). Initial manufacture of blinded IP is expected to take a minimum of 28 days. 
8.3 
Double-Blind Treatment Period 
The double-blind treatment period includes 4 scheduled clinic visits. These visits will include study 
drug administration as well as assessment of safety endpoints. 
8.3.1 Study Days 1, 30 ± 7 days, and 60 ± 7 days 
On the first study day, a 4-hour Mixed Meal Tolerance Test (MMTT) will be performed as part of 
baseline evaluations. The meal test will be performed in the morning after an overnight fast, with no 
food or drink (with the exception of water).  At the outset, a mixed meal, consisting of High Protein 
Boost will be administered.  Blood samples for C-peptide and glucose will be collected at the 
following timepoints (in minutes, relative to administration of the mixed-meal): -10, 0, 15, 30, and 
then at 30 minute-intervals until the last time point of 240 minutes.  
1-2 days prior to each study day, study personnel will contact the subject to confirm that they remain 
in generally good health and that there are no symptoms of acute illness.  
Prior to dosing on Study Day 1, a review of eligibility should be performed and confirmed. If, prior 
to dosing, a randomized subject is not suitable (ineligible) for the trial, that subject will be considered 
a “randomization failure” and will be replaced. 
The following procedures will be performed prior to study drug administration on each dosing day: 
 Any changes to medical history including prior treatment, current medicines, and drug 


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allergies, should be recorded. 
 Focused physical exam including blood pressure, pulse, temperature and weight. 
 Laboratory studies including:  
o CMP 
o CBC w diff 
o HbA1c 
o Urine MicroAlbumin panel 
o CrCl (calculation)  
  
 Measurement of auto-antibodies, including: 
o Insulin associated aAb (IAA) 
o Insulinoma associated protein 2 (IA-2, previously known as ICA-512) 
o Glutamic acid decarboxylase (GAD65) 
o Zinc transporter 8 (ZnT8). 
 A urine pregnancy test will be collected for females of child-bearing potential. 
 Any severe hypoglycemic events occurring since the prior study visit will be recorded.  
Note that non-severe events are not being collected in the database in this study, but will 
be monitored by the investigator, consistent with the site’s standard of care. 
Study drug will be administered during these visits.  Laboratory data will be collected prior to 
initiation of the infusion of study drug. 
Subjects will be observed for at least 2 hours post-end of each infusion.  Safety and tolerability 
assessments will be performed during and following the infusion, including measurement of vital 
signs.  Vital sign frequency peri-infusion is monitored per the site’s standard of care and a 
recommendation for the minimal frequency is described in the MOP.  Local tolerability at the 
infusion site (such as pain, swelling, tenderness, and erythema) will be assessed via adverse event 
reporting.  Any adverse events or new concomitant medications will be recorded. 
8.3.2 Study Day 90 ± 7 days 
Subjects will return one month after the last infusion for safety and tolerability assessments. This will 
include a physical exam, insulin usage, vital signs, clinical laboratories CMP, CBC w diff, HbA1c, 
Urine MicroAlbumin panel, CrCl (calculated), adverse events, and concomitant medications.  Local 
tolerability at the infusion site (such as pain, swelling, tenderness, and erythema) will be assessed via 
adverse event reporting.  Samples for the potency assay and other glycemic measures will be 
collected.   
 Measurement of auto-antibodies, including: 
o Insulin associated aAb (IAA) 
o Insulinoma associated protein 2 (IA-2, previously known as ICA-512) 
o Glutamic acid dexarboxylase (GAD65) 
o Zinc transporter 8 (ZnT8). 
8.4 
Long-Term Follow-Up Period 


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Continued data collection will be performed during the long-term follow-up period.  There are 5 
planned in-clinic visits during this period. 
8.4.1 Study Day 150 ± 7 days 
This visit will include a full assessment of safety, tolerability, and efficacy.  This will include a 
physical exam, insulin usage, vital signs, clinical laboratories CMP, CBC w diff, HbA1c, Urine 
MicroAlbumin panel, CrCl (calculated) adverse events, and concomitant medications.  A urine 
pregnancy test will be collected for females of child-bearing potential. 
Samples for the potency assay and for pancreatic β-cell function (MMTT) and other glycemic 
measures will be collected.   
 MMTT C-Peptide 
 Measurement of auto-antibodies, including: 
o Insulin associated aAb (IAA) 
o Insulinoma associated protein 2 (IA-2, previously known as ICA-512) 
o Glutamic acid dexarboxylase (GAD65) 
o Zinc transporter 8 (ZnT8). 
 Total daily insulin usage over the prior 3-7 days in the immediate period prior to the visit  
 Any severe hypoglycemic events occurring since the prior study visit will be recorded. 
8.4.2 Study Day 270 ± 14 days, 360 ± 14 days, 540 ± 14 days, and 720/Early Term ± 14 days 
These visits will include a physical exam (can be symptom-driven except at Study Day 360, in which 
a full physical examination is required), insulin usage, vital signs, clinical laboratories CMP, CBC w 
diff, HbA1c, Urine MicroAlbumin panel, CrCl (calculated), adverse events, and concomitant 
medications.  A urine pregnancy test will be collected for females of child-bearing potential at the 
Study Day 720/Early Term visit only. 
Samples for the potency assay will be collected, and samples for pancreatic β-cell function will be 
collected.   
 MMTT C-Peptide (MMTT will not be performed at the Day 270 visit, but will be 
collected at Day 360, 540, and 720/Early Term visits). 
 Measurement of auto-antibodies, including: 
o Insulin associated aAb (IAA) 
o Insulinoma associated protein 2 (IA-2, previously known as ICA-512) 
o Glutamic acid decarboxylase (GAD65) 
o Zinc transporter 8 (ZnT8). 
 Total daily insulin usage over the 3-7 days in the immediate period prior to the visit  
 Any severe hypoglycemic events occurring since the prior study visit will be recorded. 
8.5 
ECGs 
All scheduled ECGs will be performed after the subject has rested supine for approximately 5 
minutes. ECGs will be collected during Screening and Day 90 ± 7 days 
8.6 
MMTT Description 


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The 4-hour Mixed Meal Tolerance Test (MMTT) will be performed as a measure of beta cell 
function at multiple visits during the study. The MMTT will be performed in the morning after an 
overnight fast, with no food or drink (with the exception of water).  At the outset, a mixed meal, 
consisting of High Protein Boost will be administered.  Blood samples for C-peptide and glucose will 
be collected at the following timepoints (in minutes, relative to administration of the mixed-meal): -
10, 0, 15, 30, and then at 30 minute-intervals until the last time point of 240 minutes. An MMTT 
protocol is provided to the investigational centers that describes the procedures for performing the 
MMTT, including preparation and sampling times.   
8.7 
Prior and Concomitant Therapy and Insulin Usage 
Study participants will continue all regularly prescribed medications, except for any diabetes 
medications other than insulin.  For subjects who are already on diabetes medications and for whom 
it is clinically appropriate to discontinue, they will do a wash off period of at least 3 weeks prior to 
initiation of leukapharesis.  The introduction of systemic immunosuppressive agents, however, will 
constitute grounds for withdrawal from the study.  However, topical creams (e.g., steroidal) and 
inhaled steroids without large systemic absorption are allowed.  
Insulin usage will be recorded, using the insulin log, 3-7 days, each day, prior to visit separately from 
prior/concomitant medication therapies.  Insulin usage will include description of the daily total dose 
and regimen. Additional instructions for use are provided on the insulin log. 
8.8 
Immunoassessment “Potency” Assay Procedure 
“Potency” Assay will be performed by the sponsor with the subject blood sample after the AVT 
treatment (or placebo) to test if the function of Qa-1/HLA-E restricted CD8+ T cell is restored. 
Please see “Schedule of Events” for the dates/frequency.  The visit-specific blood volumes as well as 
specimen collection, handling, and transport procedures are described in the MOP. 
“Potency” Assay will be also performed for “information only” with AVT001 made by DFCI-CMCF 
before applying to the subject. 2-3 vials of frozen ATVT001 (1x106 cells/vial) should be sent to the 
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9 
Statistical Methodology 
A separate document entitled “Statistical Analysis Plan” (SAP) will describe further details about the 
analyses to be conducted in this study.  An overview of the general statistical methods is provided in 
the following sections. 
9.1 
Analysis Timing and Blinding Considerations 
Consistent with the study design, the primary time point for analysis is Month 5.  The primary 
analysis for this study will be performed after the last subject has completed the Month 5 study visit, 
at which time the study database will be locked and the treatment allocation codes unblinded for 
analysis.  This analysis will form the primary basis for the assessment of the study objectives. 
Continuing data collection through the long-term follow-up will be performed through Month 24.  
While Investigators and subjects will remain blinded to the treatment allocations, assessment and 
analysis of outcomes will be performed by the Sponsor in an on-going manner, to better understand 
the longer-term effects (on both safety and efficacy) of the therapy. 
9.2 
Sample Size Determination 
The primary objective of this study is to assess the safety and tolerability of AVT001, as compared to 
placebo, in subjects with type 1 diabetes.  A sample size of 14 subjects treated with AVT001 will 
provide at least 80% power to observe at least one occurrence of treatment-emergent adverse events 
with a true underlying incidence of 12%.  The sample size provides the same power to detect at least 
one occurrence of local tolerability events relating to the i.v. administration of study drug.   
Secondary objectives of this study include assessment of changes in pharmacodynamic markers, 
including (increases) in the CD8+ T-cell reg system, C-peptide levels, and HbA1c.  The sample size 
of 14 subjects treated with AVT001 and 7 subjects treated with placebo will provide estimates of the 
mean and standard deviation of the treatment benefit of AVT001 as compared to placebo in these 
endpoints.   
To account for an estimated 10% potential dropout prior to the completion of the Month 5 
safety/efficacy assessment post-first dose, a total sample size of approximately 24 subjects will be 
randomized, in a 2:1 ratio of AVT001:Placebo.  
9.3 
Analysis Populations 
There are two analysis populations defined for this study.   
 The Safety population is defined as all subjects who receive at least one dose of study 
medication.  Safety outcomes will be assessed using the Safety population. 
 Pharmacodynamic outcomes will be assessed in subjects with at least one post-baseline 
assessment for the evaluation of the CD8+ T-cell reg system biomarker.   
Other analysis populations may be defined in the SAP. 
9.4 
Baseline characteristics, Patient Disposition and Medication 
Key baseline characteristics will be tabulated by treatment group.  Patient disposition will be 
described, including patient completion status and reasons for premature discontinuation from the 
study.  Prior and concomitant medications will be tabulated (with a focus on new-onset medications, 
ie, those that are started after study treatment has commenced).   
9.5 
Adverse Events 


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Adverse events will be reported using the MedDRA coding dictionary.  Tabulations will include an 
overall incidence of at least one adverse event, incidence within body system, and incidence by 
preferred term.  Each subject may only contribute once (ie, first occurrence) to each of the incidence 
rates, regardless of the number of occurrences.  The incidence of adverse events will be presented as 
follows: 
 The incidence of treatment-emergent adverse events (TEAEs), ie, those events that start 
on or after the first dose of study medication, will be tabulated by treatment for the 
Treatment Phase.  This will include events starting within 30 days following the last dose. 
 The incidence of post-treatment events (those events that start more than 30 days 
following the last dose) will be tabulated. 
Subjects with serious adverse events (including deaths) and subjects who discontinue due to adverse 
events will be listed.  Subjects who have other significant serious adverse events deemed to be of 
special interest because of clinical importance will also be listed.   
9.6 
Severe Hypoglycemic Events and Insulin Log 
The number and subject incidence of severe hypoglycemic events (defined in Section 11.1.7) will be  
tabulated by subject incidence (number and percent of subjects) as well as frequency (number of 
events per subject).    
The total daily dose of insulin will be tabulated by visit from the insulin logs.  Daily insulin usage 
will be listed from these logs. 
9.7 
Clinical Laboratories, ECGs, and Vital Signs 
Descriptive statistics at Baseline, on-treatment, and change from Baseline will be calculated for the 
subjects who have both Baseline and on-treatment evaluations. Shifts from Baseline to on-treatment 
will be examined.  Potentially clinically significant laboratory values will be identified and listed. 
9.8 
Pharmacodynamic Outcomes 
Changes over time in CD8+ T-cell reg system, C-peptide levels, HbA1c, and antibody values will be 
assessed.  For purposes of data presentation, where appropriate, a mixed model for repeated 
measures (MMRM) will be used to present changes in these parameters over time.  Details regarding 
this MMRM will be provided in the SAP.   
9.8.1 CD8+ T-cell reg system 
A “potency assay” has been established to measure the specific suppression (inhibition) of the CD8+ 
regulatory T-cells to the target cells as one parameter to evaluate the efficacy of the AVT001 
treatment.  The data of maximum suppression of CD8+ regulatory T-cells from treated T1D 
subjects before the treatment versus after the treatment, with healthy people as a system control, 
determines whether there is a regained suppressive function of the CD8+ regulatory T-cells 
following the AVT001 treatment.  Such values might suggest the effectiveness of 
treatment.  Analyses for these outcomes will be specified in the SAP. 
9.8.2 C-Peptide AUC Calculation  
The area under the stimulated C-peptide curve (AUC) over the first 4-hour period of a mixed meal 
glucose tolerance test will be conducted at baseline and Months 5, 12, 18, and 24. The AUC at each 
visit is computed using the trapezoidal rule that is a weighted sum of the C-peptide values over the 
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transformed using the function: log(C-peptide +1). This provides better normal distributional 
behavior of the random variable.   
9.8.3 HbA1c, Antibodies 
Changes from baseline in HbA1c and antibodies will be tabulated. 


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10 Risks and Discomforts 
The Investigator’s Brochure is to be reviewed by the Investigator prior to initiation of this study.  A 
brief summary of potential risks and discomforts is provided in this section.  The drug handling 
manual includes additional detail as well as rescue medications that may be required.  
10.1 Venipuncture 
Subjects may experience slight discomfort at the site and rarely develop an ecchymosis or superficial 
thrombosis. 
10.2 Leukapheresis 
The most common adverse reactions from leukapheresis are paraesthesias, dizziness, mild chest 
tightness, or cough related to the reinfusion of autologous red blood cells treated with citrate and 
ecchymoses at the catheter sites. 
Potential adverse reactions related to leukapheresis include: 
 Exacerbation of symptoms from an underlying medical condition including migraine 
headache, asthma/emphysema, or cardiovascular disease. 
 Infection at catheter sites 
 Hypotension, hypertension, or bradycardia from citrate toxicity 
 Allergic reactions 
 Anemia, if red blood cells cannot be reinfused 
10.3 Response to AVT001 
The response to AVT001 would be to activate HLA-E restricted CD8+ T cells in vivo, which would 
in turn selectively down-regulate any anti-self responses to control autoimmune diseases.  In pre-
clinical animal toxicology studies, the data showed no adverse effect/reaction to the treatment.  In 
general, DC-based therapies are considered safe and tolerable therapies, with less toxicity 
comparable to checkpoint inhibitors in use or in trial. 
There might be infusion site reactions such as swelling and irritation after the treatment of AVT001. 
In addition, there may be risks/discomforts involved in this study that are not known at this time. 
 
 


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11 Safety and Adverse Events 
The primary objective of this study is to assess the safety and tolerability of AVT001, as compared to 
placebo, in subjects with type 1 diabetes.   
11.1 Definitions 
11.1.1 Unanticipated Problems Involving Risk to Subjects 
Any incident, experience, or outcome that meets all of the following criteria: 
 Unexpected in nature, severity, or frequency (i.e. not described in study-related 
documents such as the IRB-approved protocol or consent form, the investigators 
brochure, etc.) 
 Related or possibly related to participation in the research (i.e. possibly related means 
there is a reasonable possibility that the incident experience, or outcome may have been 
caused by the procedures involved in the research) 
 Suggests that the research places subjects or others at greater risk of harm (including 
physical, psychological, economic, or social harm). 
11.1.2 Adverse Event 
Adverse event means any untoward medical occurrence associated with the use of a drug in humans, 
whether or not considered drug related. Adverse events may include safety findings considered to be 
clinically significant by the Investigator.  An adverse drug event can therefore be any unfavorable 
and unintended sign (including an abnormal laboratory finding), symptom, or disease temporally 
associated with the use of a medicinal product, whether or not it is related to the medicinal product.  
Reporting an adverse event does not necessarily reflect a conclusion by the Investigator that the 
event is causally related to the drug. 
AE is any symptom, sign, illness or experience that develops or worsens in severity during the course 
of the study. Intercurrent illnesses or injuries should be regarded as adverse events. Abnormal results 
of diagnostic procedures are considered to be adverse events if the abnormality: 
 results in study withdrawal 
 is associated with a serious adverse event 
 is associated with clinical signs or symptoms 
 leads to additional treatment or to further diagnostic tests 
 is considered by the investigator to be of clinical significance 
A preexisting condition is one that is present at the start of the study. A preexisting condition should 
be recorded as an adverse event if the frequency, intensity, or the character of the condition worsens 
during the study period. 
Each adverse event in this study will be assessed for Grade, where Grade of an AE refers to the 
severity of the AE.  Grade will be assessed according to CTCAE Version 4.03 or higher.  The 
CTCAE displays Grades 1 through 5 with unique clinical descriptions of severity for each AE that 
are based on this general guideline, with higher grades indicating worse severity.   
The relationship of any AE to study treatment will be assessed using the following definitions: 
Definite (must have all 4) 


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 Has a reasonable temporal relationship to the intervention  
 Could not have readily been produced by the subject's clinical state or have been due to 
environmental or other interventions  
 Follows a known pattern of response to intervention  
 Disappears or decreases with reduction in dose or cessation of intervention and recurs 
with re-exposure 
Probable (must have 3) 
 Has a reasonable temporal relationship to the intervention  
 Could not have readily been produced by the subject's clinical state or have been due to 
environmental or other interventions  
 Follows a known pattern of response to intervention  
 Disappears or decreases with reduction in dose or cessation of intervention 
Possible (must have 2) 
 Has a reasonable temporal relationship to the intervention  
 Could not have readily been produced by the subject's clinical state  
 Could not readily have been due to environmental or other interventions  
 Follows a known pattern of response to intervention 
Unlikely (must have 2) 
 Does not have a temporal relationship to the intervention  
 Could readily have been produced by the subject's clinical state  
 Could have been due to environmental or other interventions  
 Does not follow a known pattern of response to intervention  
 Does not reappear or worsen with reintroduction of intervention 
Unrelated 
 The temporal relationship between treatment exposure and the adverse event is 
unreasonable or incompatible and/or adverse event is clearly due to extraneous causes 
(e.g., underlying disease, environment) 
11.1.3 Serious Adverse Event (SAE) 
Adverse events are classified as serious or non-serious. A serious adverse event is any AE that is: 
 Fatal 
 Life-threatening 
 Requires or prolongs hospital stay 
 Results in persistent or significant disability or incapacity 
 A congenital anomaly or birth defect 
 An important medical event 
Any adverse event that results in hospitalization or prolonged hospitalization should be documented 
and reported as a serious adverse event unless specifically instructed otherwise in this protocol. Any 


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condition responsible for surgery should be documented as an adverse event if the condition meets 
the criteria for an adverse event. 
Neither the condition, hospitalization, prolonged hospitalization, nor surgery are reported as an 
adverse event in the following circumstances: 
 Hospitalization or prolonged hospitalization for diagnostic or elective surgical 
procedures for a preexisting condition. Surgery should not be reported as an outcome of 
an adverse event if the purpose of the surgery was elective or diagnostic and the outcome 
was uneventful. 
 Hospitalization or prolonged hospitalization required to allow efficacy measurement for 
the study. 
 Hospitalization or prolonged hospitalization for therapy of the target disease of the study, 
unless it is a worsening or increase in frequency of hospital admissions as judged by the 
clinical investigator. 
All adverse events that do not meet any of the criteria for serious event should be regarded as non- 
serious adverse events. 
11.1.4 General Physical Examination Findings 
At screening, any clinically significant abnormality should be recorded as a preexisting condition. At 
the end of the study, any new clinically significant findings/abnormalities that meet the definition of 
an adverse event must also be recorded and documented as an adverse event.   
Physical examinations can be symptom-driven except at Screening, Month 5, and Month 12, during 
which full examinations are required. 
11.1.5 Post-study Adverse Event 
All unresolved adverse events should be followed by the investigator until the events are resolved, 
the subject is lost to follow-up, or the adverse event is otherwise explained. At the last scheduled 
visit, the investigator should instruct each subject to report any subsequent event(s) that the subject, 
or the subject’s personal physician, believes might reasonably be related to participation in this study. 
The investigator should notify the study sponsor of any death or adverse event occurring at any time 
after a subject has discontinued or terminated study participation that may reasonably be related to 
this study. The sponsor should also be notified if the investigator should become aware of the 
development of cancer or of a congenital anomaly in a subsequently conceived offspring of a subject 
that has participated in this study. 
11.1.6 Abnormal Laboratory Values 
A clinical laboratory abnormality should be documented as an adverse event if any one of the 
following conditions is met: 
 The laboratory abnormality is not otherwise refuted by a repeat test to confirm the 
abnormality 
 The abnormality suggests a disease and/or organ toxicity 
 The abnormality is of a degree that requires active management; e.g. change of dose, 
discontinuation of the drug, more frequent follow-up assessments, further diagnostic 
investigation, etc. 


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11.1.7 Severe Hypoglycemia 
Subject self-monitoring of blood glucose will be performed according to the standard of care as 
indicated by the investigator.  This may include clinician-prescribed glucose monitoring devices, ie, 
glucose meters or continuous glucose monitoring devices (CGM).  The number of severe  
hypoglycemic events will be collected as part of each study visit.  Events are defined  according to 
American Diabetes Association recommended criteria.   
 Severe hypoglycemia denotes severe cognitive impairment requiring external assistance for 
recovery. Severe hypoglycemia is an event requiring assistance of another person to actively 
administer carbohydrates, glucagon, or take other corrective actions. Plasma glucose 
concentrations may not be available during an event, but neurological recovery following the 
return of plasma glucose to normal is considered sufficient evidence that the event was 
induced by a low plasma glucose concentration. 
 In addition to the above criterion, for subjects using CGM, readings below the <54 mg/dL 
(3.0 mmol/L) for at least 15 minutes will be considered severe hypoglycemia 
Note that non-severe hypoglycemic events will not be collected or recorded in the study database, but 
that the investigator will monitor those types of events according to the site’s standard of care. 
11.2 Recording of Adverse Events 
At each contact with the subject, the investigator must seek information on adverse events by specific 
questioning and, as appropriate, by examination. Information on all adverse events should be 
recorded immediately in the source document, and also in the appropriate adverse event module of 
the case report form (CRF). All clearly related signs, symptoms, and abnormal diagnostic procedures 
results should recorded in the source document, though should be grouped under one diagnosis. 
All adverse events occurring during the study period must be recorded. The clinical course of each 
event should be followed until resolution, stabilization, or until it has been determined that the study 
treatment or participation is not the cause. Serious adverse events that are still ongoing at the end of 
the study period must be followed up to determine the final outcome. Any serious adverse event that 
occurs after the study period and is considered to be possibly related to the study treatment or study 
participation should be recorded and reported immediately. 
11.3 Reporting of Adverse Events 
11.3.1 Adverse Event Reporting Period 
The study period during which adverse events must be reported is defined as the period from the time 
of consent through the end of study or early termination, whichever is sooner.  Reporting of Serious 
Adverse Events – Study Sponsor Notification by Investigator 
A serious adverse event must be reported to the study sponsor by telephone within 24 hours of the 
event. A Serious Adverse Event (SAE) form must be completed by the investigator and faxed to the 
study sponsor within 24 hours.  The investigator will keep a copy of this SAE form on file at the 
study site.  Report serious adverse events by phone, facsimile or email to: 
Name: Dr. Harvey Katzeff, MD  
Address: Iqvia Biotech, 1700 Perimeter Park Drive, Morrisville, NC  
Phone:  919.313.7111 (US Toll-free:  1-866-758-2798) 


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Fax: 
919.313.1412 (US Toll-free:  1-866-761-1274) 
Email: Safety-Inbox@novellaclinical.com 
At the time of the initial report, provide as much of the following information as possible: 
 Study identifier 
 Study Center 
 Subject number 
 A description of the event 
 Date of onset 
 Current status 
 Whether study treatment was discontinued 
 The reason why the event is classified as serious 
 Investigator assessment of the association and expectedness between the event and study 
treatment 
 If the treatment blind was broken. 
Within the following 48 hours, the investigator must provide further information on the serious 
adverse event in the form of a written narrative. This should include a copy of the completed Serious 
Adverse Event form, and any other diagnostic information that will assist the understanding of the 
event. Significant new information on ongoing serious adverse events should be provided promptly 
to the study sponsor. 
11.3.2 IRB Notification 
Each site’s local policy for IRB reporting should be followed.  All serious adverse events (SAEs) 
should be submitted to the Institutional Review Board/Independent Ethics Committee (IRB/IEC) 
within the timing of the local authority.  The investigator should make an effort to obtain all hospital 
medical records including discharge summary confirming final diagnosis.   
The death of a subject must be immediately (within 24 hours) reported to the IRB.  Other Reportable 
Events 
For clinical drug trials, the following events are also reportable to the IRB: 
 Any adverse experience that, even without detailed analysis, represents a serious 
unexpected adverse event that is rare in the absence of drug exposure (such as 
agranulocytosis, hepatic necrosis, Stevens-Johnson syndrome). 
 Any adverse event that would cause the sponsor to modify the investigators brochure, 
protocol or informed consent form, or would prompt other action by the IRB to assure 
protection of human subjects. 
 Information that indicates a change to the risks or potential benefits of the research, in 
terms of severity or frequency. For example: 
o An interim analysis indicates that participants have a lower rate of response to 
treatment than initially expected. 
o Safety monitoring indicates that a particular side effect is more severe, or more 
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o A paper is published from another study that shows that an arm of your research 
study is of no therapeutic value. 
 Change in FDA safety labeling or withdrawal from marketing of a drug, device, or 
biologic used in a research protocol. 
 Breach of confidentiality 
 Change to the protocol taken without prior IRB review to eliminate apparent immediate 
hazard to a research participant. 
 Incarceration of a participant when the research was not previously approved under 
Subpart C and the investigator believes it is in the best interest of the subject to remain on 
the study. 
 Complaint of a participant when the complaint indicates unexpected risks or the 
complaint cannot be resolved by the research team. 
 Protocol violation (meaning an accidental or unintentional deviation from the IRB 
approved protocol) that in the opinion of the investigator placed one or more participants 
at increased risk, or affects the rights or welfare of subjects. 
11.3.3 FDA Notification by Sponsor 
The study sponsor is required to report certain study events in an expedited fashion to the FDA. 
These written notifications of adverse events are referred to as IND safety reports. The following 
describes the safety reporting requirements by timeline for reporting and associated type of event: 
 
Within 7 calendar days 
Any study event that is: 
1. associated with the use of the study drug 
2. unexpected, 
3. fatal or life-threatening, and 
 
Within 15 calendar days 
Any study event that is: 
– associated with the use of the study drug, 
– unexpected, and 
– serious, but not fatal or life-threatening 
-or- 
– a previous adverse event that was not initially deemed reportable but is later found to 
fit the criteria for reporting (reporting within 15 calendar days from when event was 
deemed reportable). 
Any finding from tests in laboratory animals that: 
– suggest a significant risk for human subjects including reports of mutagenicity, 
teratogenicity, or carcinogenicity. 
Sponsors are also required to identify in IND safety reports all previous reports concerning similar 
adverse events and to analyze the significance of the current event in light of the previous reports. 


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11.4 Protocol Deviations 
The investigator will maintain a protocol deviation log, in accordance with the site’s local policy for 
IRB reporting, that describes departures from the study protocol. If the impact on the protocol 
disrupts the study design, may affect the outcome (endpoints) or compromises the safety and welfare 
of the subjects, the deviation will be reported to the Sponsor (and the IRB in accordance with the 
site’s local policy) within 10 business days. 
Any departure from the protocol that meets the following criteria should be submitted to the Sponsor: 
 May/can/have affects/affected subject safety 
 Violate eligibility 
 Dose adjustment 
 Stopping criteria 
 Affect sample size (adding more subjects, decreasing number of subjects, changing the 
number of subject in a specific arm/cohort) 
11.5 Medical Monitoring 
It is the responsibility of the Principal Investigators to oversee the safety of the study at their sites. 
This safety monitoring will include careful assessment and appropriate reporting of adverse events as 
noted above, as well as the construction and implementation of a site data and safety-monitoring  
plan. The Sponsor will be responsible for performing overall medical monitoring, including regular 
assessment of safety outcomes (e.g., review of laboratory data, review of  adverse events and local 
tolerability outcomes across the sites). 


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12 Data Handling and Record Keeping 
12.1 Confidentiality 
Information about study subjects will be kept confidential and managed according to the 
requirements of the Health Insurance Portability and Accountability Act (HIPAA). Those regulations 
require a signed subject authorization informing the subject of the following: 
 What protected health information (PHI) will be collected from subjects in this study 
 Who will have access to that information and why 
 Who will use or disclose that information 
 The rights of a research subject to revoke their authorization for use of their PHI. 
In the event that a subject revokes authorization to collect or use PHI, the investigator, by regulation, 
retains the ability to use all information collected prior to the revocation of subject authorization. For 
subjects that have revoked authorization to collect or use PHI, attempts should be made to obtain 
permission to collect at least vital status (i.e. that the subject is alive) at the end of their scheduled 
study period. 
12.2 Source Documents 
Source data is all information, original records of clinical findings, observations, or other activities in 
a clinical trial necessary for the reconstruction and evaluation of the trial. Source data are contained 
in source documents Examples of these original documents, and data records include: hospital 
records, clinical and office charts, laboratory notes, memoranda, subjects’ diaries or evaluation 
checklists, pharmacy dispensing records, recorded data from automated instruments, copies or 
transcriptions certified after verification as being accurate and complete, microfiches, photographic 
negatives, microfilm or magnetic media, x-rays, subject files, and records kept at the pharmacy, at 
the laboratories, and at medico-technical departments involved in the clinical trial. 
12.3 Case Report Forms 
Clinical study data will be captured / collected in a clinical study database using electronic case 
report forms (eCRFs) in a CFR Title 21 part 11-compliant system. All data requested in the eCRF 
must be recorded, and reasons for missing data should be provided.  
12.4 Records Retention 
It is the investigator’s responsibility to retain study essential documents according to regulatory 
requirements.  These documents may be retained for a longer period if required by an agreement with 
the sponsor. In such an instance, it is the responsibility of the sponsor to inform the 
investigator/institution as to when these documents no longer need to be retained. 


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13 Study Monitoring, Auditing, and Inspecting 
13.1 Study Monitoring Plan 
This study will be monitored according to the study monitoring plan. The investigator will allocate 
adequate time for such monitoring activities. The Investigator will also ensure that the monitor or 
other compliance or quality assurance reviewer is given access to all the above noted study-related 
documents and study related facilities (e.g. pharmacy, diagnostic laboratory, etc.), and has adequate 
space to conduct the monitoring visit. 
13.2 Auditing and Inspecting 
The investigator will permit study-related monitoring, audits, and inspections by the EC/IRB, the 
sponsor, government regulatory bodies, and University compliance and quality assurance groups of 
all study related documents (e.g. source documents, regulatory documents, data collection 
instruments, study data etc.). The investigator will ensure the capability for inspections of applicable 
study-related facilities (e.g. pharmacy, diagnostic laboratory, etc.). 
Participation as an investigator in this study implies acceptance of potential inspection by 
government regulatory authorities and applicable University compliance and quality assurance 
offices. 


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14 Ethical Considerations 
This study is to be conducted according to US and international standards of Good Clinical Practice 
(FDA Title 21 part 312 and International Conference on Harmonization guidelines), applicable 
government regulations and Institutional research policies and procedures. 
This protocol and any amendments will be submitted to a properly constituted independent Ethics 
Committee (EC) or Institutional Review Board (IRB), in agreement with local legal prescriptions, for 
formal approval of the study conduct. The decision of the EC/IRB concerning the conduct of the 
study will be made in writing to the investigator and a copy of this decision will be provided to the 
sponsor before commencement of this study. The investigator should provide a list of EC/IRB 
members and their affiliate to the sponsor. 
All subjects for this study will be provided a consent form describing this study and providing 
sufficient information for subjects to make an informed decision about their participation in this 
study. This consent form will be submitted with the protocol for review and approval by the EC/IRB 
for the study. The formal consent of a subject, using the EC/IRB-approved consent form, must be 
obtained before that subject is submitted to any study procedure. This consent form must be signed 
by the subject or legally acceptable surrogate, and the investigator-designated research professional 
obtaining the consent. 


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Table 3  Schedule of Events (Protocol AVT001-T1D-01) 
 
 
Double-Blind Study Period (Visits 3-7) 
Open-Label Study Period (Visits 8-11) 
Study Period 
Screen/Rand/Leuk 
Double-Blind Treatment Period 
Safety Assessment 
Long-Term Follow-Up Period 
Study Day 
≤ -902 
* 
13 
30±7 
60±7 
90±7 
150±7 
 270±7 
360±7 
540±7, 720±7 / Early Term
Study Month 
-3 
* 
- 
1 
2 
3 
5 
9 
12 
18, 24 
Visit Name 
1 
2 
3 
4 
5 
6 
7 
8 
9 
10, 11 
Informed Consent
X
Vein Check
X
Randomization
X
Medical History4
X
X
Physical Exam5 
X
X
X
X
X
X
X
X
X
Insulin Usage
X
X
X
X
X
X
X
X
Vital Signs6
X
X
X
X
X
X
X
X
X
MMTT for C-peptide 
X
X
X
X
Severe Hypoglycemic Events 
Assessment
 
 
X 
X 
X 
X 
X 
X 
X 
X 
HbA1c and Antibody Samples
X
X
X
X
X
X
X
X
X
Laboratory Studies7
X
X
X
X
X
X
X
X
X
ECGs
X
X
Viral Studies8
X
Urine Pregnancy test
X
X
X
X
X
X (Day 720 /ET only)
Local tolerability of infusion site 
 
 
X 
X 
X 
X 
 
 
 
 
Leukapheresis
X9
Potency Assay10
X (BD)
X (AVT)
X (BD) 
X (BD) X (BD) 
X (BD) 
X (BD) 
Study Drug Administration11
X
X
X
Adverse Events
On-going
Prior/Conmeds
Prior Meds
Conmeds
 
2 Screening assessments may be performed at more than 1 visit, if needed, and must be completed prior to randomization 
3 At Study Day 1, a review of Medical history, symptom-driven PE, and review of eligibility should be performed prior to treatment. Details are included in note 4. 
4 Medical history including prior treatment, current medicines, and drug allergies 
5 Physical exam including blood pressure, pulse, temperature, weight.  Can be symptom-driven physical examinations, except at Screening, Month 5, and Month 12.  
6 Vital sign frequency peri-infusion is determined by site’s standard of care; recommended (minimal) guidelines are provided in the MOP 
7 Specified at each study visit: generally includes CMP, CBC w diff, Urine MicroAlbumin panel, CrCl (calculated); eGFR (calculated at screening only for eligibility) 
8 HIV-1, HIV-2, HCV serology and HBV surface antigen 
9 Leukapheresis to be performed after randomization, with sufficient time for IP/placebo manufacture prior to end of window between Screening Visit (Visit 1) and first infusion (Visit 3). Initial manufacture of blinded IP is expected to take a 
minimum of 28 days. 
10 Blood draw (BD) or AVT001 (AVT) product (collection volumes and directions as per MOP) 
11 Although visit windows allow a ±7 day window around visits, Study Drug administration must be at least 21 days between administration of double-blind study medication. 


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Jiang, H., Zhang, S.I., and Pernis, B. 1992. Role of CD8+ T cells in murine experimental 
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Liu, F., Qi, Y., Wang, X., Yang, J.J., Yang, J., Huang, X., Chess, L., and Jiang, H. 2013. 
Establishment Of A Long-term Donor Specific Transplantation Tolerance By Biologically 
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antigen presentation and induction of CD8+ cytotoxic T-cell responses in autoimmune 
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Rewers, M., and Gottlieb, P. 2009. Immunotherapy for the prevention and treatment of type 
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Giannoukakis, N., B. Phillips, D. Finegold, J. Harnaha, and M. Trucco. 2011. Phase I 


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(Safety) study of autologous tolerogenic dendritic cells in type 1 diabetes patients. Diabetes 
Care 34:2026-2032. 
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Machen, J., Harnaha, J., Lakomy, R., Styche, A., Trucco, M., and Giannoukakis, N. 2004. 
Antisense oligonucleotides down-regulating costimulation confer diabetes-preventive 
properties to nonobese diabetic mouse dendritic cells. Journal of Immunology 173:4331-
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Timmerman, J.M., Czerwinski, D.K., Davis, T.A., Hsu, F.J., Benike, C., Hao, Z.M., Taidi, 
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Bedrosian, I., Mick, R., Xu, S.W., Nisenbaum, H., Faries, M., Zhang, P., Cohen, P.A., 
Koski, G., and Czerniecki, B.J. 2003. Intranodal administration of peptide-pulsed mature 
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36. 
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A., Maczek, C., Schreiner, D., von den Driesch, P., et al. 1999. Vaccination with Mage-3A1 
peptide-pulsed mature, monocyte-derived dendritic cells expands specific cytotoxic T cells 
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E.G., Blume, K.G., and Levy, R. 1999. Idiotype vaccination using dendritic cells after 
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Perambakam, S., Hallmeyer, S., Reddy, S., Mahmud, N., Bressler, L., DeChristopher, P., 


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16 Declarations of Sponsor and Investigator 
 
Protocol Title: A Phase 1 / 2 Double-Blind, Randomized, Placebo Controlled Study of Safety, 
Tolerability and Potential Efficacy of AVOTRES Cell-Based Therapy (AVT001) in Patients with 
Type 1 diabetes 
 
Protocol Number: AVT001-T1D-01 
 
Protocol Version/ Date: Amendment 3 Dated 1Feb2021 
 
Sponsor Name: Avotres Inc. 
 
16. 1. Declaration of Sponsor 
The above study protocol was reviewed and has been approved by the appropriate protocol review 
committee of the Sponsor.  The information it contains is consistent with: 
 The current risk-benefit evaluation of the IP; 
 The moral, ethical, and scientific principles governing clinical research as set out in the 
Declaration of Helsinki and the principles of ICH guidelines for GCP and applicable local 
regulatory requirements. 
The Investigator will be supplied with details of any significant or new findings, including adverse 
events (AE), related to the IP. 
 
Sponsor  
Signature:  ______________________________________ Date:_____________ 
Name (print):  Hong Jiang, MD/Ph.D., Chief Scientific Officer, Avotres Inc. 
 
16. 2. Declaration of Investigator 
I confirm that I have read the above protocol.  I understand it, and I will work according to the 
protocol and moral, ethical, and scientific principles governing clinical research as set out in the 
Declaration of Helsinki and the principles of ICH guidelines for GCP and according to applicable 
local regulatory requirements. 
 
Investigator (Dr. Jason Gaglia) 
 
Signature:  ______________________________________ Date:_____________ 
Name (print):  Jason Gaglia, MD, MMSc 
Institution name: Joslin Diabetes Center 
Institution address: 1 Joslin Place, Boston, MA 02215 
                            
 


Amendment 1 Summary of Changes for Protocol AVT001-T1D-01 
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Page 1 of 18 
 
 
 
 
AMENDMENT 1: SUMMARY OF CHANGES FOR 
PROTOCOL AVT001-T1D-01 
 
Investigational Product: 
 
AVT001 Suspension for Intravenous Infusion  
Hsp60sp-loaded Autologous Immature Dendritic Cells 
(7x106-10x106cells/dose)  
 
Study Title: 
A Phase 1 / 2 Double-Blind, Randomized, Placebo Controlled 
Study of Safety, Tolerability and Potential Efficacy of 
AVOTRES Cell-Based Therapy (AVT001) in Patients with Type 
1 Diabetes 
 
 
This document summarizes the changes from the original protocol 
(dated July 15, 2013) to Amendment 1 (dated January 28, 2019). 
 
Amendment 1 maintains the same general study design as the original protocol.  Modifications and 
clarifications have been made in Amendment 1, included both structural/formatting and content 
updates.  The only key content updates are in reference to additions to the safety monitoring (eg, the 
addition of vital signs, ECGs), and further description of the DSMB reviews.  There have been no other 
meaningful changes to the study design or procedures as described in the original protocol. 
Because the structural changes to the format of the document resulted in renumbered the section 
headers, the first colum of the table in the following pages provides the Section Titles for the amended 
document only.  This column also includes a brief description of the key change being described. 
The second column includes the text/language from the original protocol. 
The third column includes the revised/new language from the amended protocol. 


Amendment 1 Summary of Changes for Protocol AVT001-T1D-01 
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The intent in this Summary of Changes document is not to provide documentation of each and every 
change (as the number of minor edits was substantial and would interfere with the clarity of this 
summary).  Rather, the intent of this Summary of Changes is intended to communicate the key 
modifications in a clear, succinct and practical fashion.  
 


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Page 3 of 18 
 
 
Section Number / Section Title 
Original Protocol 
Amendment 1 
All sections 
 
The protocol document has had extensive 
formatting, structural and stylistic updates 
made throughout to better align with ICH and 
FDA guidances.  The section numbers have 
also been updated and modified. 
 
Much of the detailed chemistry, manufacturing 
and control (CMC) and dosing procedures 
have been shifted to a manual of operating 
procedures (MoP) and allow for the removal of 
sections such as reconciliation and disposition 
processes. 
Section 3:  Synopsis 
A “Study Summary” was provided. 
An ICH-compliant synopsis has replaced the 
original ‘Study Summary’ table. 
 
Synopsis and protocol components have been 
aligned (e.g. objectives, endpoint, and 
inclusion / exclusion criteria.) 
Section 3: (PI/Study Center) 
 
The first study center has been 
changed.  Additional centers may be 
added as enrolment needs dictate. 
Subjects will be recruited for this study from the clinical 
practices of the Columbia University Medical Center and 
its referral sources. Subjects will be required to give written 
informed consent to participate in the study before any 
screening tests or evaluations are conducted. 
The first study center is now the Joslin 
Diabetes Center, Boston, with Jason Gaglia, 
MD, MMSc as the PI. However, an additional 
center may be added as enrollment and accrual 
needs dictate. 
Section 4: Introduction 
 
Minor updates to the background, disease 
description, and study treatment description 
have been made.  AVT001 product name has 
been established. 


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Section Number / Section Title 
Original Protocol 
Amendment 1 
Section 5: Study Objectives and 
Design 
 
The study objectives have been better 
aligned to reflect the primary focus of 
the study on safety and tolerability.  
Pharmacodynamic outcomes have 
been defined as secondary. 
 
These changes are aligned with the 
synopsis as well.  
Primary Objectives 
– To determine the safety and to Tolerability of the 
AVOTRES reagent. 
– To determine whether, among T1D subjects with in 
vitro-reversible defects in the CD8+ T cell regulatory 
system, infusion of AVOTRES will lead to an in vivo 
reconstitution of this system. 
Secondary Objectives 
– To determine whether, among T1D subjects with in vitro-
reversible defects in the CD8+ T cell regulatory system, 
infusion of AVOTRES will result in amelioration of β cell 
destruction. 
Primary Outcomes 
• To determine the safety and tolerability 
profile of three administrations of a single 
dose  AVT001 (as compared to placebo) in 
T1D subjects with in vitro-reversible 
defects in the CD8+ T cell regulatory 
system  
Secondary Outcomes 
• To determine whether, among T1D subjects 
with in vitro-reversible defects in the CD8+ 
T cell regulatory system, infusion of 
AVT001 will lead to an in vivo 
reconstitution of this system. 
• To determine whether, among T1D subjects 
with in vitro-reversible defects in the CD8+ 
T cell regulatory system, infusion of 
AVT001 will result in β cell preservation 
assessed by stimulated C-peptide. 
Section 5.3: Study Design and 
Section 5.4:  Study Duration 
The duration of the study was defined as 24 months, with an 
additional 2 years of ‘follow-up’ to assess survival/disease 
status. 
The duration of the study has been revised via 
the definition of three ‘periods’ being defined 
for the study, ie, Screening, Treatment Period 
(through Month 3), and Post-Treatment 
Follow-up Period (through Month 12).  Thus, 
the primary time point for assessment in the 
study is now Month 5, the duration of the 
‘follow-up’ portion study has been shortened 
to extend out to 12 months post-first dose, and 
the longer-term followup may occur through 
up to Month 24.  


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Section Number / Section Title 
Original Protocol 
Amendment 1 
Section 5.5: Subject Enrollment 
 
The number of patients subject to 
staggered enrollment prior to open 
enrollments is reduced from 9 to 6. 
 
Reducing the number of staggered 
patients: Given the latest evidence 
and track record of safety with 
dendritic cell therapies since the 
initial filing of this IND (references 
1-9), and the fact that in this study 
there is no dose escalation and we are 
evaluating a single concentration 
range of AVT001 therapy for each 
subject, we are proposing to reduce 
the number of staggered subjects to 6 
individuals, separated by 4 weeks 
each. We believe that this, in 
continued oversight of unblinded data 
by the DSMB, will ensure adequate 
safety monitoring during the course 
of the trial.   
 
NOTE:  No specific language in the protocol.  Instead, the 
staggered enrollment applies as follows: 
 
We will monitor the first 9 patients to make sure that at least 
6 subjects in the treatment group will be observed for 4 weeks 
for acute and subacute adverse events.  The information 
obtained will allow the clinical PI and Sponsor to determine if 
the time interval of 2 weeks or 4 weeks is sufficient, we will 
consult with the FDA to lengthen or shorten the 4-week 
observation of patient staggering period in the clinical trial. 
 
See attached: 
 
 
 
 
 
Subject exposure will be managed using a 
staggered enrollment that will allow for 
review of each subjects outcomes following 
the initial IV.  This staggered enrollment will 
be in-place for the first 6 subjects treated.   
 
Subjects treated at a rate of one subject per 
month, with Investigator and Medical Monitor 
review of the Month 1 safety and tolerability 
outcomes for each subject before treatment of 
the subsequent subject is performed.  
 


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Section Number / Section Title 
Original Protocol 
Amendment 1 
Section 5.6: DSMB 
The Data Safety and Monitoring Board (DSMB) will monitor 
the data quality and adherence to safety rules. Additionally, 
the DSMB will review all safety/toxicity data for the trial and 
recommend trial suspension or termination as needed. 
Specific details of monitoring and audit frequency will  be  
included in the Monitoring Plan (see Appendix A)  but will be 
at least every 6 months 
 
Investigators will report all SAEs to the DSMB within 10 
business days regardless of expectedness or attribution. SAE 
will be input into the CRF with follow up email to DSMB. 
A DSMB will be established that will monitor 
subject safety as well as data quality and 
adherence to safety rules. Additionally, the 
DSMB will review all safety/toxicity data for 
the trial and recommend trial modifications, 
suspension or termination, as needed.  
 
The DSMB will initially meet after 3 
subjects and then after 6 subjects have 
received at least one dose of double-blind 
study medication and have completed their in-
clinic visit through at least one month post-
dose.  The DSMB will review unblinded data 
to assess the safety and tolerability of the 
treatments given.  At the second meeting (in 
which 6 subjects are being evaluated), the 
DSMB will also review preliminary efficacy 
data (in an unblinded fashion).  The DSMB 
will, following each of these meetings, provide 
the Sponsor with a summary of their findings 
and recommendations (if any) for protocol 
amendments.   
 
The interval of subsequent meetings of the 
DSMB will be determined by the DSMB Chair 
and Sponsor.  A DSMB Charter will be 
written, to include details regarding meeting 
frequency, format, and membership. 
 


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Section Number / Section Title 
Original Protocol 
Amendment 1 
5.7:  Study Endpoints 
Primary Study Endpoints 
(a) 
Safety and tolerability of the AVOTRES reagent. The 
primary analysis will describe the incidence of adverse events 
and   laboratory abnormalities. 
(b) 
Tolerability of the AVOTRES reagent. 
(c) 
Activity of the HLA-E-restricted CD8+ T cell 
regulatory function over a 1-year period following treatment 
with AVOTRES. 
 
Secondary Study Endpoints 
(a) 
The change in the area under the curve (AUC) of the 
stimulated C-peptide levels over the first 2 hours of a mixed 
meal tolerance test (MMTT) 3 months, 6 months, 12 months, 
18 months, and 24 months 
(b) 
Change in A1c 
(c) 
Change in total daily insulin dose 
(d) 
Number of major hypoglycemic events (defined as 
loss of consciousness, seizure, or requiring assistance from 
another person because of altered state of consciousness) 
(e) 
Reported hypoglycemic events confirmed with 
capillary blood glucose measurement less than 70 mg/dl. 
(f) 
Autoantibody levels 
The study endpoints have been re-ordered to 
better reflect the primary and secondary 
objectives fo the study.  Thus, the primary 
endpoints are related only to safety and 
tolability, while the secondary endpoints are 
related to the pharmacodynamic objectives. 
 
The primary endpoints are: 
(a) The incidence of treatment-emergent 
adverse events (TEAEs).   
(b) Clinically significant changes from 
baseline in clinical laboratory parameters.  
(c) The incidence and severity of local IV-site 
reactions.  
(d) Changes from baseline in vital signs and 
electrocardiograms.  
(e) Incidence of severe hypoglycemic events.  
Secondary endpoints will include: 
(a) Assessment of the HLA-E-restricted 
CD8+ T cell regulatory activity 
(“potency assay”) 
(b) Changes from baseline in the area under 
the curve (AUC) of the stimulated C-
peptide levels over the first 4 hours of a 
mixed meal tolerance test (MMTT) 
(c) Changes from baseline in HbA1c 
(d) Change from baseline in insulin usage 
(e) Changes from baseline in autoantibody 
levels 


A me n d me nt 1 S u m mar y of C ha n ges f or Pr ot oc ol A V T 0 0 1- T 1 D - 0 1 
Ja n uar y 2 8, 2 0 1 9 Versi o n 1  
A V T 0 0 1:  A V O T R E S Cell -Base d T hera p y   
 
A v otres, I nc. 
Pa ge 8  of 1 8  
 
 
Secti o n 6:  S u bject Selecti o n 
(I ncl usi o n / E xcl usi o n Criteri a) 
 
T he sa me u n derl yi n g st u d y 
p o p ulati o n has bee n retai ne d.  Mi n or 
c ha n ges t o i n di vi d ual i ncl usi o n a n d 
e xcl usi o n criteria ha ve bee n ma de t o 
better -defi ne t he eli gi bilit y 
re q uire me nts. 
 
Re m o ve d i ns uli n prere q uisite fr o m 
ori gi nal versi o n. 
 
Re m o vi n g t he i ns uli n t hera p y 
re q uire me nt: T he criteri o n f or 
re q uire me nt of i ns uli n t hera p y per 
trial was re vise d t o defer t o t he 
pri mar y treati n g p h ysicia n t o pr o vi de 
sta n dar d of care i ns uli n t hera p y, i n 
or der t o pr o vi de ma xi mal fle xi bilit y 
f or t he treati n g healt hcare pr o vi ders.   
 
S u bjects w h o meet all of t he f oll o wi n g criteria are eli g i ble f or 
e nr oll me nt as st u d y partici pa nts:  
Dia g n osis of T y pe 1 Dia betes b y o ne or m ore of t he f oll o wi n g 
t y pes of a nti b o dies: a nti- G A D 6 5 ( gl uta mic aci d 
decar b o x ylase), a nti – islet-cell a nti b o d y 5 1 2 (I C A 5 1 2), a nti -
I A 2 a n d a nti-i ns uli n a nti b o d y.s u bjects 
1. 
A ge  1 8 or ol der a n d a ble t o pr o vi de i nf or me d c o nse nt 
f or partici pati o n. 
2. 
M ust ha ve sti m ulate d C - pe pti de le vels 0. 2 p m ol/ ml 
meas ure d d uri n g a mi xe d meal t olera nce test ( M M T T)  
3.  
s u bjects S u bjects Rece nt o nset ( dia g n osis wit hi n t he 
past 1 2 m o nt hs) T 1 D s u bjects  w h o scree n p ositi ve f or a 
C D 8 + T cell defect t hat is c orrecta ble i n vitr o b y c o- c ult ure 
wit h i m mat ure de n dritic cells l oa de d wit h self - pe pti de 
Hs p 6 0s p. 
4. 
All s u bjects will be usi n g i ns uli n at t he ti me of 
e nr oll me nt. 
5. 
If a partici pa nt is fe male wit h re pr o d ucti ve p ote ntial, 
s he m ust be willi n g t o a v oi d pre g na nc y a n d ha ve a ne gati ve 
pre g na nc y test d uri n g t he 1 2 m o nt hs of treat me nt a n d f or a n 
a d diti o nal 3 m o nt hs. 
6. 
Si g ne d a n d date d writte n i nf or me d c o nse nt  
 
 
S u bjects w h o meet a n y of t he f oll o wi n g criteria are n ot 
eli gi ble f or e nr oll me nt as st u d y partici pa nts:  
 
1.  
S u bjects S u bjects S u bjects He m o gl o bi n < 1 0. 0 g/ d L; 
le u k oc ytes < 3, 0 0 0/ u L; ne utr o p hils < 1, 5 0 0/ u L; l y m p h oc ytes 
< 8 0 0/ u L; platelets < 1 0 0, 0 0 0/ u L 
2.  
Uri ne Al b u mi n E xcreti o n < 3 0 0 m g/ g m Cr 
3.  
e G F R > 6 0 m L/ mi n/ 1. 7 3 m 2  
4. 
A L T/ A S T < 3 x n or mal  
5.  
C urre nt use of i m m u n os u p pressi ve or 
i m m u n o m o d ulat or y t hera pies, i ncl u di n g p har mac ol o gic d oses 
of s yste mic ster oi ds  
6.  
C oi nci de nt me dical c o n diti o n li kel y t o re q uire 
i m m u n os u p pressi ve or i m m u n o m o d ulat or y t hera pies 
6. 1  
I ncl usi o n Criteri a 
S u bjects w h o meet all of t he f oll o wi n g criteria 
are eli gi ble f or e nr oll me nt as st u d y 
partici pa nts:  
1.  Di a g n osis of t y pe 1 di a betes, wit hi n t he 
p ast 1 2 m o nt hs b ut at le ast 1 m o nt h 
pri or t o scree ni n g , c o nfir me d b y 
p ositi ve la b res ult f or o ne or m ore of t he 
f oll o wi n g t y pes of a ut oa nti b o dies: 
a. Gl uta mic aci d decar b o x ylase 
( G A D 6 5) 
b.  I ns uli n o ma ass ociate d pr otei n 2 
(I A- 2, als o k n o w n as I C A- 5 1 2) 
c. Zi nc tra ns p orter 8 ( Z n T 8).  
2.  A ge 1 8 or ol der a n d a ble t o pr o vi de 
i nf or me d c o nse nt. 
3.  S u bjects w h o scree n p ositi ve f or a 
C D 8 + T cell defect t h at is c orrect a ble 
i n vitr o b y c o-c ult ure wit h i m mat ure 
de n dritic cells l oa de d wit h t he s y nt hetic 
oli g o pe pti de fr o m Hs p 6 0s p.  
4.  If a partici pa nt is fe male wit h re pr o d ucti ve 
p ote ntial, s he m ust be willi n g t o a v oi d 
pre g na nc y a n d ha ve a ne gati ve pre g na nc y 
test t hr o u g h t he d urati o n of t he trial.  
5.  Si g ne d a n d date d writte n i nf or me d 
c o nse nt. 
 
6. 2 
E xcl usi o n Criteri a  
S u bjects w h o meet a n y of t he f oll o wi n g 
criteria are n ot eli gi ble f or e nr oll me nt as st u d y 
partici pa nts:  
1.  
S u bjects o n e xisti n g treat me nt wit h 
p o orl y c o ntr olle d gl yce mic c o ntr ol 
des pite i ns uli n t hera p y, w h o i n t he 


Amendment 1 Summary of Changes for Protocol AVT001-T1D-01 
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7. 
Coincident medical condition likely to limit short 
term (5 year) life expectancy (malignancy, symptomatic 
coronary artery disease, recent stroke) 
8. 
Prior radiation therapy, immunotherapy, or 
chemotherapy 
9. 
Serologic evidence of current HIV-1 or HIV-2 
infection 
10. 
Evidence of current hepatitis B as demonstrated by 
HBSAg or circulating hepatitis B genomes 
11. 
Serologic evidence of hepatitis C infection 
12. 
SubjectsAutoimmune disease aside from T1D 
13. 
Pregnant or breastfeeding 
14. 
Any chronic illness which, in the opinion of the 
principal investigator, should preclude participation in the 
trial 
15. 
Adequate venous access to support leukapheresis 
opinion of the investigator would not be 
a good candidate for participation in a 
clinical trial 
2. 
Screening hemoglobin <10.0 g/dL; 
leukocytes <3,000/uL; neutrophils 
<1,500/uL; lymphocytes <800/uL; 
platelets <100,000/uL 
3. 
Screening Urine Albumin Excretion > 
300 mg/gmCr 
4. 
Screening eGFR < 60 mL/min/1.73m
2
 
5. 
Screening ALT or AST > 1.5x upper 
limit of normal (ULN) 
6. 
Screening bilirubin > 2.0 mg / dL 
7. 
Current use of immunosuppressive or 
immunomodulatory therapies, including 
pharmacologic doses of systemic 
steroids.  However, topical steroidal 
creams and inhaled steroids without 
large systemic absorption are allowed. 
8. 
Coincident medical condition likely to 
require immunosuppressive or 
immunomodulatory therapies. 
9. 
Coincident medical condition likely to 
limit short term (5 year) life expectancy 
(malignancy, symptomatic coronary 
artery disease, recent stroke) 
10. 
Prior radiation therapy, immunotherapy, 
or chemotherapy 
11. 
Serologic evidence of current HIV-1 or 
HIV-2 infection 
12. 
Serologic evidence of hepatitis C 
infection 
13. 
Serologic evidence of acute or chronic 
active hepatitis B as measured by Core 


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Section Number / Section Title 
Original Protocol 
Amendment 1 
Ab positive and / or Surface Antibody 
antigen positive  
14. 
Subjects with non-T1D autoimmune 
conditions (except autoimmune thyroid 
and/or celiac diseases) 
15. 
Women who are pregnant, 
breastfeeding, or planning pregnancy 
during the study period. 
16. 
Inadequate venous access to support 
leukapheresis. 
17. 
Any condition that in the opinion of the 
investigator(s) would preclude the 
subject from participating in a clinical 
trial. 
18. 
Abnormal baseline ECG that in the 
opinion of the investigator or sponsor 
would pose a safety risk. 
 


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Section 6.3 Early Withdrawal of 
Sujects 
Criteria for Premature Termination from the Study 
Subjects may withdraw from the study prior to the expected 
completion date for the following reasons: 
• 
Unacceptable toxicity and other safety reasons 
• 
Progression or disease relapse 
• 
Subject consent withdrawal 
• 
Decision by the investigators that withdrawing is 
in the patient’s best interest 
• 
Death 
Standard supportive therapy will be maintained for subjects 
withdrawn from active treatment.  
Data Collection and Follow-up of Withdrawn 
ubjects 
Every effort will be made to collect toxicity information on 
withdrawn subjects.  Subjects  withdrawn prematurely from 
the trial will be followed-up for survival and disease status by 
the principal investigator or clinical investigators via clinic 
visit, telephone contact, or medical records review every 6 
months for 2 years. Other methods that will be used for 
follow-up will include contacting next of kin, contacting 
referring or primary physicians, use of certified letters to the 
patient, and social security index. 
 
If a patient is discontinued from the study 
prematurely, the Investigator must select the 
primary reason for discontinuation on the End 
of Study eCRF.  In addition, every effort 
should be made to complete the assessments 
listed under the End of Treatment column on 
the Schedule of Events.  
 
Patients withdrawn from the study will be 
considered evaluable for statistical assessment. 
 
A patient may be removed from the study for 
the following medical or administrative 
reasons: 
• 
Adverse Event: If a patient experiences an 
adverse event that the patient finds 
unacceptable or that, in the judgment of 
the Investigator or the Medical Monitor 
presents an unacceptable consequence or 
risk to the patient, the patient may be 
discontinued from further participation in 
the study. 
• 
Administrative Discontinuation:  After 
consultation with the Investigator or 
Medical Monitor, a patient may be 
discontinued from the study for failure to 
comply with protocol requirements.  All 
instances of noncompliance must be 
documented in the eCRF. 
• 
Refusal of Assessments:  If for any reason, 
following dosing, the patient refuses 
further assessment during the study, the 
patient shall be discontinued from the 
study and the reasons for refusal 
documented.  Reasonable efforts shall be 
made to monitor the patient for adverse 
events following such discontinuation.  


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Section Number / Section Title 
Original Protocol 
Amendment 1 
Such efforts shall be documented. 
 
Standard supportive therapy will be 
maintained for subjects withdrawn from active 
treatment.   Every effort will be made to 
collect safety data from withdrawn subjects.   
Section 7: Description of Study 
Drug 
 
Updated section in alignment with CMC 
procedures of drug supply including selection 
of Dana Farber Cancer Institute Cell 
Manufacturing Core Facility (DFCI-CMCF) 
Section 8:  Study Procedures 
 
The entire Study Procedures section 
has been updated to better-define the 
evaluations to be performed at each 
visit.   
 
The visit timing and assessments at 
each visit have been modified to 
allow for better and more meaningful 
data collection that is consistent with 
the study objectives and design. 
See the original Schedule of Events at the end of this 
document 
See the amended Schedule of Events at the 
end of this document.  More details have been 
provoed regarding the procedures to be 
performed at each study visit. 
 
Additional safety assessments have been 
added, including vital signs and ECGs 
 
Visits have been added (and removed) to 
match the study design. 
 
MMTT Description has been added 
 
Many supply related details have been taken 
out and be placed into a manual of operating 
procedures (MoP). 
 
Study visit ranges have been modified to 
ensure timely yet convenient windows for a 
site visit from patients to maximize adherence 
to the schedule. 
 
Assay procedure clarifications have been 
added. 


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Section Number / Section Title 
Original Protocol 
Amendment 1 
Section 9:  Statistical Methodology 
 
The statistical methods sections have 
been rewritten to better reflect the 
timing of the analysis, to describe the 
analysis populations, and details as to 
how safety and tolerability data will 
be analyzed. 
 
The entire Statistical Methods section has been 
rewritten and now better matches the study 
design and objectives. 
Section 9.1:  Sample Size 
Determination 
 
The sample size methodology has 
been updated to reflect the primary 
objective of the study (safety and 
tolerability). 
Analysis assumes that the proportion of subjects given 
placebo exhibiting reversal at 12 months will be 0 and that at 
least 60% of the subjects receiving AVOTRES will exhibit in 
vivo evidence of reversal of the defect. With a randomization 
of 2:1 subjects to Avotres or Placebo. N = 14 subjects 
randomized to Avotres and N = 7 subjects to placebo. 
There would be 94% power to detect this difference in 
proportions of 0.6 with a one sided α <0.05 using the Fisher’s 
exact test. Allowing for up to 10% of subjects  being 
unavailable for evaluation the total sample size is  increased  
to N = 24. 
The primary objective of this study is to assess 
the safety and tolerability of AVT001, as 
compared to placebo, in subjects with type 1 
diabetes.  A sample size of 14 subjects 
treated with AVT001 will provide at least 
80% power to observe at least one occurrence 
of treatment-emergent adverse events with a 
true underlying incidence of 12%.  The sample 
size provides the same power to detect at least 
one occurrence of local tolerability events 
relating to the i.v. administration of study drug.  
 
Secondary objectives of this study include 
assessment of changes in pharmacodynamic 
markers, including (increases) in the CD8+ T-
cell reg system, C-peptide levels, and HbA1c.  
The sample size of 14 subjects treated with 
AVT001 and 7 subjects treated with placebo 
will provide estimates of the mean and 
standard deviation of the treatment benefit of 
AVT001 as compared to placebo in these 
endpoints.   
 
To account for an estimated 10% potential 
dropout prior to the completion of the Month 
5 safety/efficacy assessment post-first dose, a 
total sample size of approximately 24 subjects 
will be randomized, in a 2:1 ratio of 
AVT001:Placebo. 


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Section Number / Section Title 
Original Protocol 
Amendment 1 
Section 11: Safety and Adverse 
Events 
 
Clarification of safety endpoints has been 
added, as well as more robust description of 
safety data collection. 
Section 11.3.2: Reporting of Serious 
Adverse Events – Study Sponsor 
Notification by Investigator 
 
The information of medical monitor 
for SAE has been updated 
Name:       Hong Jiang, MD, PhD 
Address:   Columbia University Medical Center  
                  PH - 8 East, Ste. 101, 
                  622 West 168th St 
                  New York, NY 10032 
Phone:       212-305-9984/201-944-3352 
Fax:           212-305-6070 
Email:      hj4@columbia.edu 
 
 
 
 
Name:      Dr. Kristy Freeman Woods, MD,        
MPH, FACP  
Address:   Novella Clinical, 1700 Perimeter 
Park Drive, Morrisville, NC  
Phone:      919.313.7111 (US Toll-free:1-866- 
758-2798) 
Fax:          919.313.1412 (US Toll-free:1-866-
761-1274) 
Email:     Safety-Inbox@novellaclinical.com 
Table 3:  Schedule of Events 
The original schedule of events can be found at the end of this 
document. 
The amended schedule of events can be found 
at the end of this document.  Changes included 
the addition of new visits, removal of select 
visits, addition of new safety procedures 
(ECG, vital signs), and more details on safety 
data collection. 
 
 


A me n d me nt 1 S u m mar y of C ha n ges f or Pr ot oc ol A V T 0 0 1- T 1 D - 0 1 
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Pa ge 1 5  of 1 8  
 
 
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2 
V A C #
3 
F/
U 
 
F/
U 
 
F/
U 
 
F/
U 
 
F/
U 
 
F/
U 
 
S T U D Y D A Y  
-6 0  
-3 0  
0  
3 0  
6 0  
9 0  
1 2 0  
1 8 0  
3 6 5  
--- 
--- 
S T U D Y M O N T H  
-1  
0  
--- 
1  
2  
3  
4  
6  
1 2  
1 8  
2 4  
S T U D Y Y E A R  
--- 
--- 
--- 
--- 
--- 
--- 
--- 
--- 
1  
--- 
2  
Hist or y 1  
X  
 
 
 
 
X  
 
 
 
 
 
P h ysical E xa m 2  
X  
 
 
 
 
X  
 
 
X   
X  
Pa ncreatic  cell 
F u ncti o n 3  
X 
 
X 
X 
X 
X 
 
X 
X 
X 
X 
La b orat or
y 
 
X 
 
X 
X 
X 
X 
 
 
X 
 
X 
A d verse 
E ve nt 
 
X 
 
X 
X 
X 
X 
 
 
 
 
 
Viral St u dies 5  
X  
 
 
 
 
 
 
 
 
 
 
S ur vi val a n d 
Disease 
 
 
 
 
 
 
 
 
 
 
 
 
Ser u m Pre g na nc y  
X  
 
 
 
 
 
 
 
 
 
 
Le u ka p heresis  
 
X   
 
 
 
 
 
 
 
 
P ote nc y Assa y 7  
X  
X   
 
 
X  
 
X  
X  
X  
X  
 
VI SI T N A M E  
VI S
I T 
1 1  
VI S
I T 
1 2  
VI S
I T 
1 3  
VI S
I T 
1 4  
P R O C E D U R E S  
L T F U 
 
L T F U 
 
L T F U 
 
L T F U 
 
S T U D Y D A Y  
--- 
--- 
--- 
--- 
S T U D Y M O N T H  
3 0  
3 6  
4 2  
4 8  
S T U D Y Y E A R  
--- 
3  
--- 
4  
Hist or y 1  
 
 
 
 
P h ysical E xa m 2  
 
X  
 
X  


Amendment 1 Summary of Changes for Protocol AVT001-T1D-01 
January 28, 2019 Version 1 
AVT001:  AVOTRES Cell-Based Therapy 
 
Avotres, Inc. 
Page 16 of 18 
 
 
Pancreatic β cell Function3 
X 
X 
Laboratory Studies4 
X 
X 
Adverse Event Assessment 
Viral Studies5 
Survival and Disease 
6
X 
X 
X 
X 
Serum Pregnancy 
Leukapheresis 
Potency Assay7 
 
 


Amendment 1 Summary of Changes for Protocol AVT001-T1D-01 
January 28, 2019 Version 1 
AVT001:  AVOTRES Cell-Based Therapy 
 
Avotres, Inc. 
Page 17 of 18 
 
 
Amended Schedule of Events 
 
Study Period 
Screening 
Double-Blind Treatment Period 
Safety Assessment 
Long-Term Follow-Up Period 
Study Day 
-60±7 
-30±14 
11 
30±7 
60±7 
90±7 
150±7 
270±7 
360±7 
560±7, 720±7 / Early Term 
Study Month 
-2 
-1 
- 
1 
2 
3 
5 
9 
12 
18, 24 
Visit Name 
1 
2 
3 
4 
5 
6 
7 
8 
9 
10, 11 
Informed Consent 
X 
 
 
 
 
 
 
 
 
 
Vein Check 
X 
 
 
 
 
 
 
 
 
 
Randomization2 
 
X 
 
 
 
 
 
 
 
 
Medical History3 
X 
 
X 
 
 
 
 
 
 
 
Physical Exam4 
X 
 
X 
 
 
X 
X 
X 
X 
X 
Insulin Usage 
 
 
X 
X 
X 
X 
X 
X 
X 
X 
Vital Signs 
X 
 
X 
X 
X 
X 
X 
X 
X 
X 
MMTT for C-peptide  
 
 
X 
 
 
 
X 
 
X 
X 
Severe Hypoglycemic Events 
Assessment 
 
 
X 
X 
X 
X 
X 
X 
X 
X 
HbA1c and Antibody Samples 
X 
 
X 
X 
X 
X 
X 
X 
X 
X 
Laboratory Studies5 
X 
 
X 
X 
X 
X 
X 
X 
X 
X 
ECGs 
X 
 
 
 
 
 
 
 
 
 
Viral Studies6 
X 
 
 
 
 
 
 
 
 
 
 Urine Pregnancy test 
X 
 
X 
X 
X 
 
X 
 
 
X (Day 720 /ET only) 
Local tolerability of infusion site 
 
 
X 
X 
X 
X 
 
 
 
 
Leukapheresis 
 
X 
 
 
 
 
 
 
 
 
Potency Assay7 
X (BD) 7 X (AVT) 7 
 
 
 
X (BD) 7 
X (BD) 7 X (BD) 7 X (BD) 7 
X (BD) 7 
Study Drug Administration8 
 
 
X 
X 
X 
 
 
 
 
 
Adverse Events 
 
 
On-going 
Prior/Conmeds 
Prior Meds 
Conmeds 
 
 
 
1 At Study Day 1, a review of Medical history, symptom-driven PE, and review of eligibility should be performed prior to treatment. Details are included in note 4. 
2 Randomization should occur after successful completion of leukapheresis. 
3 Medical history including prior treatment, current medicines, and drug allergies 
4 Physical exam including blood pressure, pulse, temperature, weight.  Can be symptom-driven physical examinations, except at Screening, Month 5, and Month 12.  
5 CBC/differential, creatinine, AST, ALT, and total bilirubin; including creatinine clearance and albuminuria 
6 HIV-1, HIV-2, HTLV-1/2, and HCV serology and HBV surface antigen 
7 Blood draw (BD) ~80 mL, collected into Sodium Heparin blood collection tubes  or AVT001 (AVT) product   
8 Although visit windows allow a ±7 day window around visits, Study Drug administration must be at least 21 days between administration of double-blind study 
medication. 


Amendment 1 Summary of Changes for Protocol AVT001-T1D-01 
January 28, 2019 Version 1 
AVT001:  AVOTRES Cell-Based Therapy 
Avotres, Inc. 
Page 18 of 18 
References 
1.
Phillips , B.E, Garciaflgueroa, Y., Trucco, M., Glannoukakis, N. 2017. Clinical tolerogenic dendritic cells: exploring therapeutic
impact on human autoimmune disease. Frontiers in Immunology, October 201: 8, Article 1279
2.
Herbert, G., Vreeland, T., Clifton, G. , Peoples, G. 2018. Initial phase I/IIa trial results of an auatologous tumor lysate, partical-loaded,
dendritic cell (TLPLDC) vaccine in patients with solid tumors. Vaccine 36 (2018) 3247-3253
3.
Chao, T., Xiaowen, W.,  Zhiqi, L., Qi, Y., Zixiao. Y, et al. 2015. A Systemic review of clinical trials on dendritic-cells based vaccine
against malignant glioma. Journal of Carcinogenesis & Mutagenesis 2015.6:222
4.
Macatangay, B., Riddler, S., Wheeler, N., Spindler, J., Lawani, M., Hong, F., Buffo, M., Whiteside, T., Kearney, M., Mellors, J.,
Rinaldo, C. 2016. Therapeutic vaccination with dendritic cells loaded with autologous HIV type 1-infected apoptotic cells. The
Journal of Infectious Diseases 2016:213 (1 May) 1400-1409
5.
Green, J., Schneble, E., Jackson, D., Hale, D., Vreeland, T., et al. 2016. A phase I/IIa clinical trial in stage IV melanoma of
auntologous tumor-dendritic cell fusion (dendritoma) vaccine with low dose interleukin-2. Cancer Immunol Immunother (2016)
65:383-392
6.
Podrazil, M., Horvath, R., Becht, E., Rozkova, D., Bilkova, P., et al.  2015. Phase I/II clinical trial of dendritic-cell based
immunotherapy (DCVAC/PCa) combined with chemotherapy in patients with metastatic, castration-resistant prostate cacner.
www.impactjournals/oncotarget Oncotarget, Vol.6, No.20
7.
Ogasawara, M., Miyashita, M., Ota, S. 2018. Vaccination of urological cancer patients with WT1 peptide-pulsed dendritic cells in
combination with molecular targeted therapy or conventional chemotherapy induces immunological and clinical responses.
Therapeutic Apheresis and Dialysis 2018:22(3): 266-277
8.
Kim, S., Jung, H., Lee, C. 2018. Generation, characteristics and clinical trials of ex vivo generated tolerogenic dendritic cells. Yonsei
Medical Journal 2018 Sep: 59 (7): 807-815
9.
Subbiah, V., Murthy, R., Hong, D., Prins, R., Hosing, C., Hendricks, K., Kolli. D. et al. 2018. Cytokines produced by dendritic cells
administered intratumorally correlate with clinical outcome in patients with diverse cancers. Clinical caner research. July 2018. CCR-
17-2707.


Amendment 2 for Protocol AVT001-T1D-01
22Jan2020
AVT001:  Hsp60sp-loaded Autologous Immature Dendritic Cells
Avotres, Inc.
CONFIDENTIAL
Summary of Protocol AVT001-T1D-01 From Amendment No. 1 to No. 2 Changes
Below is a by-section summary of changes associated with this protocol amendment. Minor
schedule clarifications, updated versioning, as well as insignificant grammatical or format
changes may not be listed. Specific language changes are either summarized in
underlined/strikethrough text or included in the tracked changes version of the protocol
amendment and are not all entirely duplicated verbatim in this summary.
Section
Summary of Change
DSMB
Updated to include a third DSMB safety meeting and to remove the primary efficacy
evaluation.
Eligibility
Select inclusion and exclusion criteria have been updated in the protocol to ensure
consistency and clarity.  In addition, inclusion criterion # 1 has been revised to reflect a
previously approved administrative protocol change.
Investigational product,
dosage and mode of
administration
Revised to clarify that subjects randomized to AVT001 will have 3 such bags
manufactured for infusion
Duration of treatment
and Figure 1: Study
Design Schematic
Screening and cell collection period revised to up to 3 months.
Efficacy Assessment
The following additional language has been inserted:
The SAP will also detail the HLA-E restricted CD8+ regulatory T cell function analysis.
General Study Design
Removed text as indicated:
Subjects will be monitored for physical or reported symptoms as well as for vital signs
(including temperature, heart rate, blood pressure, and arterial oxygen saturation) and
changes in physical exam throughout the infusion procedure and for a minimum of 2 hours
following the procedure.
Stopping Rules
Added clarification of protocol attribution to include possibly, probably or definitely.
Early Withdrawal
of Subjects
Last sentence in section revised as indicated:
Every effort will be made to collect safety data from withdrawn these subjects through
the end of study.
Table 2:
Investigational
Product
Dosage Form for both IP and Placebo clarified to include 10% DMSO in Human
Serum Albumin
Screening/
Randomization/
Leukapheresis
Entire section revised and reordered to better describe sequencing of screening,
randomization and leukapheresis, to reflect a previously approved administrative
protocol change, to ensure consistency with the Schedule of Events, to refer the reader
to the Manual of Operations for more detailed product handling and procedures, to add
specificity of required lab and other assessments, and to add granularity around the
randomization schema.


Amendment 2 for Protocol AVT001-T1D-01
22Jan2020
AVT001:  Hsp60sp-loaded Autologous Immature Dendritic Cells
Avotres, Inc.
CONFIDENTIAL
Section
Summary of Change
Double-Blind
Treatment Period
The following additional language has been inserted:
If, prior to dosing, a randomized subject is not suitable (ineligible) for the trial, that
subject will be considered a “randomization failure” and will be replaced.
Vital sign frequency peri-infusion is monitored per the site’s standard of care and a
recommendation for the minimal frequency is described in the MOP.
Specificity of required laboratory assessments have been added/clarified.
Long-Term Follow-
Up Period
Specificity of required laboratory assessments have been added/clarified.
Prior and
Concomitant
Therapy and Insulin
Usage
The following additional language has been inserted:
Additional instructions for use are provided on the insulin log.
Immunoassessment
“Potency” Assay
Procedure
Entire section revised to read:
“Potency” Assay will be performed by the sponsor with the subject blood sample after AVT
treatment (or placebo) to test if the function of Qa-1/HLA-E restricted CD8+ T cell is
restored. Please see “Schedule of Events” for the dates/frequency.  The visit-specific blood
volumes as well as specimen collection, handling, and transport procedures are described in
the MOP.
“Potency” Assay will be also performed for “information only” with AVT001 made
by DFCI-CMCF before applying to the subject. 2-3 vials of frozen ATVT001 (1x106
cells/vial) should be sent to the sponsor for such assessment.
Baseline
characteristics,
Patient Disposition
and Medication
Entire section added as follows:
Key baseline characteristics will be tabulated by treatment group.  Patient disposition
will be described, including patient completion status and reasons for premature
discontinuation from the study.  Prior and concomitant medications will be tabulated
(with a focus on new-onset medications, ie, those that are started after study treatment
has commenced).
Severe
Hypoglycemic
Events and Insulin
Log
The following additional language has been inserted:
The total daily dose of insulin will be tabulated by visit from the insulin logs.  Daily
insulin usage will be listed from these logs.
CD8+ T-cell reg
system
Entire section added as follows:
A “potency assay” has been established to measure the specific suppression
(inhibition) of the CD8+ regulatory T-cells to the target cells as one parameter to
evaluate the efficacy of the AVT001 treatment. The data of maximum
suppression of CD8+ regulatory T-cells from treated T1D subjects before the
treatment versus after the treatment, with healthy people as a system control,
determines whether there is a regained suppressive function of the CD8+ regulatory T-
cells following the AVT001 treatment. Such values might suggest the effectiveness of
treatment. Analyses for these outcomes will be specified in the SAP.


Amendment 2 for Protocol AVT001-T1D-01
22Jan2020
AVT001:  Hsp60sp-loaded Autologous Immature Dendritic Cells
Avotres, Inc.
CONFIDENTIAL
Section
Summary of Change
Adverse Event
Reporting Period
First paragraph revised as follows:
The study period during which adverse events must be reported is defined as the
period from the time of consent through the end of study or early termination,
whichever is sooner. as the period from the initiation of dosing (Day 1) to the end of
the study treatment follow-up. For this study, the study treatment follow-up is defined
as 30 days following the last administration of study treatment.  SAEs that occur prior
to Study Day 1 should also be reported.
In addition, the medical monitor is changed from Dr. Kristy Freeman Woods to Dr.
Harvey Katzeff and Novella Clinical is changed to Iqvia Biotech.
Case Report Forms
Added that eCRFs are maintained in a CFR Title 21 part 11-compliant system.
Ethical
Considerations
Removed reference to Appendix 4 for a copy of the Subject Informed Consent Form
as this is a stand-alone document.
Table 3: Schedule
of Events
Clarified which visits are blinded and which are not.
Double-Blind Study Period (Visits 3-7)
Open-Label Study Period (Visits 8-11)
Clarified several footnotes to conform to protocol procedures, to clarify laboratory
assessments, to correct Potency Assay collection requirements, and to reflect a
previously approved administrative protocol change


Amendment 2 for Protocol AVT001-T1D-01 
 
1Feb2021 
AVT001:  Hsp60sp-loaded Autologous Immature Dendritic Cells 
Avotres, Inc. 
 
CONFIDENTIAL 
Summary of Protocol AVT001-T1D-01 From Amendment No. 2 to No. 3 Changes 
 
 
Below is a by-section summary of changes associated with this protocol amendment. Minor 
schedule clarifications, updated versioning, as well as insignificant grammatical or format 
changes may not be listed.  Specific language changes are either summarized in 
underlined/strikethrough text or included in the tracked changes version of the protocol 
amendment and are not all entirely duplicated verbatim in this summary.  
 
 
Section 
Summary of Change 
Inclusion 
Inclusion now permits participants aged 16 and older.  Assent required, where applicable. 
Exclusion 
 
Exclusion criterion for bilirubin cut-off modified to include those with Gilbert’s Syndrome 
as follows:  
Screening bilirubin > 2.0 mg / dL, or > 3.0 mg / dL for participants with Gilbert’s 
Syndrome  
Dosing Rationale 
 
The following additional language has been added for clarification around rationale for 
including older adolescents aged 16-17: 
According to a 2017–2018 National Health Interview Survey (NHIS), National Center for 
Health Statistics, and Centers for Disease Control and Prevention report, as of 2018 an 
estimated 187,000 children and adolescents younger under 20 years of age had been 
diagnosed with with type 1 diabetes (49).  Following the DSMB review of the safety data 
for the first 6 staggered adult subjects (18 years and older), no safety concerns were 
identified.  Therefore, due to the safety established thus far and due to the high prevalence 
of disease in adolescents, this protocol is being amended (Amendment # 3) in a step-wise 
fashion to reduce the lower limit of age for eligibility to age 16.  [With new reference] 
Screening  
 
Clarified that urine pregnancy testing expires after 90 days from collection (instead of 30 
days) 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 


Avotres, Inc. 
Protocol AVT001-T1D-01 
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Author: 
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Version Date:  
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STATISTICAL ANALYSIS PLAN 
AVT001-T1D-01 
A Phase 1 / 2 Double-Blind, Randomized, Placebo 
Controlled Study of Safety, Tolerability and Potential 
Efficacy of AVOTRES Cell-Based Therapy (AVT001) in 
Patients with Type 1 Diabetes
AUTHOR: MARIANA HILDESHEIM 
VERSION NUMBER AND DATE: V1.0, 28MAR2022 


Avotres, Inc. 
Protocol AVT001-T1D-01 
 
Page 2 of 39 
 
Author: 
Mariana Hildesheim 
Version Number: 
1.0 
 
 
Version Date:  
28Mar2022 
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Reference: 
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Effective Date: 02Dec2019 
 
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proprietary to IQVIA Holdings Inc. and its subsidiaries. Unauthorized use, disclosure or reproduction is strictly prohibited. 
STATISTICAL ANALYSIS PLAN SIGNATURE PAGE 
Statistical Analysis Plan v1.0 (dated 28Mar2022) for protocol AVT001-T1D-01. 
 
Name 
Signature 
Date (DDMmmYYYY) 
Author: 
Mariana Hildesheim 
 
 
Position: 
Sr Biostatistician 
Company: 
IQVIA Biotech 
Upon review of this document, the undersigned approves this version of the Statistical Analysis Plan, authorizing 
that the content is acceptable for the reporting of this study. 
 
Name 
Signature 
Date (DDMmmYYYY) 
Approved by: 
Tuochuan Dong 
 
 
Position: 
Biostatistician 
Company: 
Avotres, Inc. 
 
 
28-Mar-2022 | 21:26:26 JST
28-Mar-2022 | 07:20:51 PDT


Avotres, Inc. 
Protocol AVT001-T1D-01 
 
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Version Date:  
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proprietary to IQVIA Holdings Inc. and its subsidiaries. Unauthorized use, disclosure or reproduction is strictly prohibited. 
MODIFICATION HISTORY 
Unique 
Identifier for 
this Version 
Date of the 
Document 
Version 
Author 
Significant Changes from  
Previous Version 
0.1 
22Dec2021 
Mariana Hildesheim 
Not Applicable – First Version 
0.2 
20Jan2022 
Mariana Hildesheim 
Planned analyses revised to condense TLF 
counts 
0.3 
04Mar2022 
Mariana Hildesheim 
Sponsor review comments addressed 
0.4 
16Mar2022 
Mariana Hildesheim 
Sponsor review comments addressed 
1.0 
28Mar2022 
Mariana Hildesheim 
Final version 
 
 
 
 


Avotres, Inc. 
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Author: 
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Version Date:  
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proprietary to IQVIA Holdings Inc. and its subsidiaries. Unauthorized use, disclosure or reproduction is strictly prohibited. 
TABLE OF CONTENTS 
1. 
INTRODUCTION............................................................................................................... 8 
2. 
STUDY OBJECTIVES AND ENDPOINTS ..................................................................... 8 
2.1. 
Primary Objective and Endpoints .................................................................................................................. 8 
2.2. 
Secondary Objectives and Endpoints ............................................................................................................. 9 
2.3. 
Exploratory Objectives and Endpoints .......................................................................................................... 9 
3. 
STUDY DESIGN ............................................................................................................... 10 
3.1. 
General Description ....................................................................................................................................... 10 
3.2. 
Schedule of Events ......................................................................................................................................... 11 
3.3. 
Changes to Analyses from Protocol.............................................................................................................. 11 
4. 
PLANNED ANALYSES ................................................................................................... 11 
4.1. 
Data Safety Monitoring Board (DSMB) ...................................................................................................... 12 
4.2. 
Primary Analysis ........................................................................................................................................... 12 
4.3. 
Final Analysis ................................................................................................................................................. 12 
5. 
ANALYSIS POPULATIONS .......................................................................................... 12 
5.1. 
All Screened Subjects Analysis Population ................................................................................................. 12 
5.2. 
Safety Population ........................................................................................................................................... 13 
5.3. 
Pharmacodynamic Population ...................................................................................................................... 13 
5.4. 
Process for Analysis Population Assignment............................................................................................... 13 
6. 
GENERAL CONSIDERATIONS ................................................................................... 13 
6.1. 
Reference Start Date and Study Day ........................................................................................................... 13 


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6.2. 
Baseline ........................................................................................................................................................... 14 
6.3. 
Unscheduled Visits, Retests, and Early Termination Data ........................................................................ 14 
6.4. 
Windowing Conventions ............................................................................................................................... 14 
6.5. 
Common Calculations ................................................................................................................................... 14 
7. 
STATISTICAL CONSIDERATIONS ............................................................................ 15 
7.1. 
Sample Size Calculation ................................................................................................................................ 15 
7.2. 
Missing data ................................................................................................................................................... 15 
7.3. 
Statistical Tests .............................................................................................................................................. 15 
7.4. 
Multiple Comparisons/ Multiplicity ............................................................................................................. 15 
7.5. 
Multicenter Studies ........................................................................................................................................ 16 
7.6. 
Adjustments for Covariates and Factors to be Included in Analyses ....................................................... 16 
7.7. 
Examination of Subgroups ............................................................................................................................ 16 
7.8. 
Software Version ............................................................................................................................................ 16 
8. 
OUTPUT PRESENTATIONS ......................................................................................... 16 
9. 
DISPOSITION AND WITHDRAWALS ........................................................................ 16 
9.1. 
Disposition ...................................................................................................................................................... 17 
9.2. 
Protocol Deviations ........................................................................................................................................ 17 
10. 
DEMOGRAPHIC AND OTHER BASELINE CHARACTERISTICS ....................... 17 
10.1. Derivations ..................................................................................................................................................... 18 
11. 
SURGICAL AND MEDICAL HISTORY ...................................................................... 18 
12. 
DISEASE HISTORY ........................................................................................................ 18 


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12.1. Derivations ..................................................................................................................................................... 19 
13. 
MEDICATIONS ............................................................................................................... 19 
14. 
EXPOSURE TO STUDY DRUG ..................................................................................... 20 
14.1. Derivations ..................................................................................................................................................... 20 
15. 
COMPLIANCE WITH STUDY DRUG ......................................................................... 21 
16. 
EFFICACY ENDPOINTS................................................................................................ 21 
16.1. Primary Efficacy ............................................................................................................................................ 21 
16.2. Secondary Efficacy ........................................................................................................................................ 21 
16.2.1. 
Secondary Efficacy Endpoint Variables and derivations ......................................................................... 21 
16.2.1.1. 
Change from Baseline in the Area under the Curve (AUC) of the stimulated C-peptide levels over 
the first 4 hours of a mixed meal tolerance test (MMTT) .................................................................................... 21 
16.2.1.2. 
Change from Baseline in HbA1c ..................................................................................................... 22 
16.2.1.3. 
Change from Baseline in Total Daily Insulin Dose, Total Daily Basal Insulin Dose, and Total Daily 
Bolus Insulin Dose ............................................................................................................................................... 22 
16.2.1.4. 
Change from Baseline in Autoantibody Levels ............................................................................... 22 
16.2.2. 
Analysis of Secondary Efficacy Endpoints .............................................................................................. 22 
16.2.2.1. 
Continuous Secondary Efficacy Endpoints ...................................................................................... 22 
16.3. Exploratory Efficacy ..................................................................................................................................... 23 
16.3.1. 
Exploratory Efficacy Endpoint Variables and Derivations ...................................................................... 24 
16.3.1.1. 
Change from Baseline in unstimluated C-peptide of a MMTT........................................................ 24 
16.3.1.2. 
Change from Baseline in the change in AUC of C-peptide (Delta AUC) of a MMTT .................... 24 
16.3.1.3. 
Change from Baseline in the maximum concentration of C-peptide (Cmax) of a MMTT .............. 24 
16.3.1.4. 
Change from Baseline in the maximum change from baseline concentration (Delta Cmax) of the 
stimulated C-peptide levels of a mixed meal tolerance test (MMTT) ................................................................. 24 
16.3.1.5. 
Change from Baseline in the AUC of the stimulated glucose of a MMTT ...................................... 25 
16.3.1.6. 
Change from Baseline in unstimluated glucse of a MMTT ............................................................. 25 
16.3.1.7. 
Change from Baseline in the change in AUC of glucose (Delta AUC) of a MMTT ....................... 25 
16.3.1.8. 
Change from Baseline in the maximum concentration of glucose (Cmax) of a MMTT .................. 25 
16.3.1.9. 
Change from Baseline in the maximum change from baseline concentration (Delta Cmax) of the 
stimulated glucose levels of a MMTT ................................................................................................................. 25 
16.3.2. 
Analysis of Exploratory Efficacy Endpoints ........................................................................................... 25 
16.3.2.1. 
Continuous Exploratory Efficacy Endpoints ................................................................................... 25 
17. 
SAFETY ENDPOINTS .................................................................................................... 26 


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17.1. Adverse Events ............................................................................................................................................... 26 
17.1.1. 
All TEAEs and Post-Treatment AEs ....................................................................................................... 26 
17.1.1.1. 
CTCAE Grading for AEs ................................................................................................................. 27 
17.1.1.2. 
Relationship to Study Drug .............................................................................................................. 27 
17.1.2. 
The listing of all AEs will include a column for relatedness.Adverse Events with an Outcome of Death
 
27 
17.1.3. 
The listing of all AEs will include a column for outcome.Serious Adverse Events ................................ 27 
17.1.4. 
AEs Leading to Discontinuation of Study Drug ...................................................................................... 28 
17.1.5. 
Severe Hypoglycaemic Adverse Events .................................................................................................. 28 
17.1.6. 
Local IV-Site Reactions ........................................................................................................................... 28 
17.2. Deaths ............................................................................................................................................................. 28 
17.3. Laboratory Evaluations ................................................................................................................................ 28 
17.3.1. 
CTCAE Toxicity Grades ......................................................................................................................... 29 
17.3.2. 
Laboratory Normal Ranges ...................................................................................................................... 29 
17.4. Vital Signs ....................................................................................................................................................... 30 
17.4.1. 
Vital Signs Markedly Abnormal Criteria ................................................................................................. 30 
17.5. ECG Evaluations ........................................................................................................................................... 31 
17.5.1. 
ECG Markedly Abnormal Criteria........................................................................................................... 32 
18. 
DATA NOT SUMMARIZED OR PRESENTED .......................................................... 32 
APPENDIX 1. PARTIAL DATE CONVENTIONS ............................................................. 34 
Algorithm for Prior Adverse Events, Treatment-Emergent  Adverse Events, and Post-Treatment Adverse 
Events 34 
Algorithm for Prior / Concomitant Medications .................................................................................................... 35 
APPENDIX 2. LABORATORY ASSESSMENTS ................................................................ 36 
 
 


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1. INTRODUCTION 
This statistical analysis plan (SAP) describes the rules and conventions to be used in the presentation and analysis of 
efficacy and safety data for protocol AVT001-T1D-01. It describes the data to be summarized and analyzed, 
including specifics of the statistical analyses to be performed.  
This SAP is based on protocol amendment 3, dated 01Feb2021, inclusive of 2 administrative change letters to 
amendment 3, dated 16Jun2021 and 03Nov2021.  
2. STUDY OBJECTIVES AND ENDPOINTS 
2.1. PRIMARY OBJECTIVE AND ENDPOINTS 
Objectives  
Endpoints  
To determine the safety and tolerability profile of 
three doses of AVT001 (as compared to placebo) in 
T1D subjects within-vitro-reversible defects in the 
CD8+ T cell regulatory system. 
The incidence of treatment-emergent adverse events 
(TEAEs). 
Clinically significant changes from baseline in clinical 
laboratory parameters. 
The incidence and severity of local IV-site reactions. 
Changes from baseline in vital signs and 
electrocardiograms. 
Incidence of severe hypoglycaemic events. 
 


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2.2. SECONDARY OBJECTIVES AND ENDPOINTS 
Objectives  
Endpoints  
To determine whether, among T1D subjects with in 
vitro-reversible defects in the CD8+ T cell 
regulatory system, infusion of AVT001 will lead to 
an in vivo reconstitution of this system. 
To determine whether, among T1D subjects with in 
vitro-reversible defects in the CD8+ T cell 
regulatory system, infusion of AVT001 will result 
in amelioration of β cell destruction as assessed by 
stimulated C-peptide. 
 
Assessment of the HLA-E-restricted CD8+ T cell 
regulatory activity (“potency assay”). 
Changes from baseline in the area under the curve 
(AUC) of the stimulated C-peptide levels over the first 
4 hours of a mixed meal tolerance test (MMTT). 
Changes from baseline in HbA1c. 
Change from baseline in insulin usage. 
Changes from baseline in autoantibody levels. 
 
2.3. EXPLORATORY OBJECTIVES AND ENDPOINTS 
Although not specifically defined in the protocol, the following endpoints will be examined as exploratory. 


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Objectives  
Endpoints  
To explore the effects of infusion of AVT001 on 
stimulated and unstimulated C-peptide and glucose 
measurements made over the first 4 hours of a 
mixed meal tolerance test (MMTT). 
 
 
Changes from baseline in C-peptide and Glucose 
measured over the first 4 hours of a MMTT: 
• 
Unstimulated C-peptide 
• 
Delta AUC C-peptide 
• 
Cmax C-peptide 
• 
Delta Cmax C-peptide 
• 
Total AUC glucose 
• 
Unstimulated glucose 
• 
Delta AUC glucose 
• 
Cmax glucose 
• 
Delta Cmax glucose 
 
3. STUDY DESIGN 
3.1. GENERAL DESCRIPTION 
This study is a phase 1 / 2 double-blind, randomized, placebo-controlled study to evaluate the safety and tolerability 
of AVT001 cell-based therapy, and to assess its potential efficacy for the treatment of type 1 diabetes (T1D). A brief 
overview of the trial design is outlined in Table A. 
Table A: 
Study Design 
 
 


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3.2. SCHEDULE OF EVENTS 
Schedule of events can be found in Table 3 of the protocol. 
3.3. CHANGES TO ANALYSES FROM PROTOCOL 
Potency assay data will be listed in the final Clinical Study Report.  In addition, as an exploratory endpoint, it will 
be evaluated internally by Avotres and will be reported separately.   
4. PLANNED ANALYSES 
The following analyses will be performed for this study: 
• 
Primary analysis after the last subject completes the Month 5 study visit (after study unblinding). All 
available data will be analyzed.   
• 
Final analysis 


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4.1. DATA SAFETY MONITORING BOARD (DSMB) 
A DSMB was established to monitor subject safety as well as data quality and adherence. Data reports for review by 
the members were generated from the electronic data capture (EDC) system by the IQVIA Biotech Data 
Management team. The IQVIA Biotech unblinded randomization statistician had a copy of the randomization 
schedule on hand at the closed sessions to provide the DSMB members with unblinded treatment assignments if 
needed. 
Refer to the DSMB Charter for more details about the DSMB composition, responsibilities, and meeting frequency. 
4.2. PRIMARY ANALYSIS 
The primary analysis will take place for this study once the last subject completes the Month 5 study visit. The study 
database will be locked at this time and the treatment allocation codes unblinded for analysis. All analyses described 
in this plan will be produced. This analysis will form the primary basis for the assessment of the study objectives. 
All planned analyses identified in the SAP will be performed by IQVIA Biotech Biostatistics following Sponsor 
Authorization of this SAP, Sponsor authorization of the analysis sets, database lock (DBL), and general study 
unblinding. 
4.3. FINAL ANALYSIS  
The final analysis will take place once the last subject completes the Month 24 long-term follow-up visit. All 
analyses described in this plan will be produced. 
5. ANALYSIS POPULATIONS 
5.1. ALL SCREENED SUBJECTS ANALYSIS POPULATION 
The all screened subjects (SCRN) analysis population will contain all subjects who provide informed consent for 
this study. 


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5.2. SAFETY POPULATION 
The safety (SAF) population is defined as all subjects who receive at least one dose of study medication. If there is 
any doubt whether a subject was treated or not, he/she will be assumed treated for the purposes of analysis. 
For analyses and displays based on SAF, subjects will be classified according to the actual treatment received. 
5.3. PHARMACODYNAMIC POPULATION 
The pharmacodynamic (PD) population is defined as all subjects in the safety population with at least one post-
baseline assessment for the evaluation of the CD8+ T-cell reg system biomarker. If there is any doubt whether a 
subject was treated or not, he/she will be assumed treated for the purposes of analysis.  
For analyses and displays based on PD, subjects will be classified according to the actual treatment received. 
5.4. PROCESS FOR ANALYSIS POPULATION ASSIGNMENT 
For the PD population, the identification and agreement of protocol deviations or events which affect PD results will 
be performed between the biostatistician, and the sponsor, prior to DBL, with sponsor authorization of any excluded 
subjects or their data. 
6. GENERAL CONSIDERATIONS 
6.1. REFERENCE START DATE AND STUDY DAY 
Study Day will be calculated from the reference start date and will be used to show start/stop day of assessments and 
events. Reference start date is defined as the day of the first injection of study drug i.e., Day 1. 
Study Day will be computed as follows: 
• 
Study Day = (Date of event –Date of first injection of study drug) + 1 if the date of the event is on or after the 
date of the first injection of study drug 
• 
Study Day = (Date of event – Date of first injection of study drug) if the date of the event is prior to the date of 
the first injection of study drug 


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In the situation where the event date is partial or missing, Study Day and any corresponding durations will appear 
partial or missing in the listings. 
6.2. BASELINE 
Unless otherwise specified, baseline is defined as the last non-missing measurement taken prior to the first injection 
of study drug (including unscheduled assessments). In the case where the last non-missing measurement and the date 
and time of the first injection of study drug coincide, that measurement will be considered pre-baseline, but adverse 
events (AEs) and medications commencing on the date and time of the first injection of study drug will be 
considered post-baseline. 
6.3. UNSCHEDULED VISITS, RETESTS, AND EARLY TERMINATION 
DATA 
For by-visit summaries, data recorded at the nominal visit will be presented. That is, unscheduled, retest (same visit 
number assigned), and early termination measurements will not be included in by-visit summaries but might 
contribute to the baseline timepoint and/or maximum value, where required (e.g. shift table).  
Listings will include scheduled, unscheduled, retest and early discontinuation data. 
6.4. WINDOWING CONVENTIONS  
No visit windowing for analysis purposes will be performed for this study.  
6.5. COMMON CALCULATIONS 
Change from baseline will be calculated as: 
• 
Change from baseline = Test value at post-baseline visit – Baseline value 
Percent change from baseline will be calculated as: 
• 
Percent change from baseline (%) = (Change from baseline at post-baseline visit / Baseline value) * 100% 
For select parameters where data is available, change and percent change will also be calculated from diagnosis. 


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7. STATISTICAL CONSIDERATIONS 
For continuous data, descriptive statistics (i.e., n [number of subjects with available data], mean, standard deviation 
[SD], median, 25th and 75th percentiles, and minimum, and maximum values) will be presented by treatment group 
and visit, when applicable. 
For categorical data, the number and percentages of subjects in each category will be presented by treatment group 
and visit, when applicable 
7.1. SAMPLE SIZE CALCULATION 
The sample size of 14 subjects treated with AVT001 will provide at least 80% power to observe at least one 
occurrence of treatment-emergent adverse events with a true underlying incidence of 12%. The sample size provides 
the same power to detect at least one occurrence of local tolerability events relating to the i.v. administration of 
study drug. 
To account for an estimated 10% potential dropout prior to the completion of the Month 5 safety/efficacy 
assessment post-first dose, a total sample size of approximately 24 subjects will be randomized, in a 2:1 ratio of 
AVT001:Placebo. 
7.2. MISSING DATA 
Missing efficacy data will not be imputed.  
Missing safety data will not be imputed, except for missing AE severity and relationship data (refer to Section 
17.1.1). Partial or completely missing AE and medication dates will be handled as described in APPENDIX 1. 
7.3. STATISTICAL TESTS 
All statistical tests will be conducted at the two-sided 5% significant level, unless otherwise specified in the 
description of the analyses.  Confidence Intervals (CIs) will be two-sided with 95% coverage. 
7.4. MULTIPLE COMPARISONS/ MULTIPLICITY 
No adjustment for multiple comparisons and multiplicity will be performed.  Only nominal p-values will be 


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provided for the efficacy endpoints. 
No statistical testing will be performed for the safety endpoints. 
7.5. MULTICENTER STUDIES 
This study will be conducted by one investigator at one center. Pooling of data for analysis is not applicable.  
7.6. ADJUSTMENTS FOR COVARIATES AND FACTORS TO BE 
INCLUDED IN ANALYSES 
 Mixed-effect Model for Repeated Measurements (MMRM) analyses C-peptide total AUC will be adjusted for 
treatment, HbA1c, total insulin, BMI, potency assay, study visit, and treatment by study visit interaction. Section 
16.2 for further details. 
7.7. EXAMINATION OF SUBGROUPS 
No subgroup analyses are planned.  
7.8. SOFTWARE VERSION 
All analyses will be conducted using SAS version 9.4.  
8. OUTPUT PRESENTATIONS 
The presentation of data in outputs will follow IQVIA Biotech Data Display Standards. Refer to the TFL shells 
document for additional information. 
9. DISPOSITION AND WITHDRAWALS 
All subjects who provide informed consent will be accounted for in this study.  


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9.1. DISPOSITION 
Number of subjects screened will be presented overall for the SCRN analysis population.  Number and percentage 
of subjects with screen failure and reason for screen failure will also be presented overall based on the SCRN 
analysis population.  Number of subjects randomized will be presented overall and by treatment group for the SCRN 
analysis population. 
Number and percentages of subjects treated, who completed/discontinued early from treatment (including reason for 
withdrawal), and who completed/discontinued early from the study (including reason for withdrawal) will be 
provided overall and by treatment group based on the SAF analysis population. 
A listing showing inclusion and exclusion of each subject from each analysis population, including reason for 
exclusion, will be provided. 
9.2. PROTOCOL DEVIATIONS 
A listing of protocol deviations identified by the study team (important or not) will be provided. 
10. DEMOGRAPHIC AND OTHER BASELINE CHARACTERISTICS 
The following demographic and other baseline characteristics collected during the screening visit will be reported 
for this study: 
• 
Age (years) – calculated relative to date of consent 
• 
Age groups 
• 
Sex 
• 
Childbearing potential (for female subjects only) 
• 
Race 
• 
Ethnicity 
• 
Weight (kg) 
• 
Height (cm) 


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• 
Body Mass Index (BMI) (kg/m2) 
Continuous demographic and other baseline characteristics will be summarized using descriptive statistics overall 
and by treatment group based on the SAF analysis population.  For categorical demographic and other baseline 
characteristics, number and percentage of subjects in each category will be provided overall and by treatment group 
based on the SAF analysis population.  No statistical testing will be carried out for demographic or other baseline 
characteristics. 
10.1. DERIVATIONS 
BMI, in kg/m2, will be calculated as follows:  
• 
BMI (kg/ m2) = weight (kg)/ [height (m)2] 
11. SURGICAL AND MEDICAL HISTORY 
Surgical and medical history are defined as any surgeries that happened before the first injection of study drug and 
any medical conditions/diseases that started and stopped before the first injection of study drug.  
Surgical and medical history will be coded using the Medical Dictionary for Regulatory Activities (MedDRA), 
version 22.0, and will be summarized by System Organ Class (SOC) and Preferred Term (PT) overall and by 
treatment group based on the SAF analysis population.  A subject having more than one surgery/medical 
condition/disease within the same SOC/PT will be counted only once for that SOC or PT. 
All surgical and medical history will be listed. 
12. DISEASE HISTORY 
The following disease history characteristics will be summarized overall and by treatment group based on the SAF 
analysis population: 
• 
Time since diagnosis (days) – calculated relative to date of first infusion of study drug 
• 
Use of insulin pump (yes/no) 
• 
HbA1c at diagnosis, screening, and baseline (%) 
• 
Change and percent change in HbA1c from diagnosis to baseline (%) 


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• 
Weight at diagnosis, screening, and baseline (kg) 
• 
Change and percent change in weight from diagnosis to baseline (kg) 
• 
BMI at diagnosis, screening, and baseline (kg/m2) 
• 
Change and percent change in BMI from diagnosis to baseline (kg/m2) 
All disease history characteristics will be listed. 
12.1. DERIVATIONS 
‘Time since’ disease history characteristics, in days, will be calculated as follows: 
• 
Time since diagnosis (days) = (Date of first infusion of study drug - Date of diagnosis) 
Change from diagnosis will be calculated as follows: 
• 
Change from diagnosis = Test value at screening or first dosing visit – Diagnosis value 
Percent change from baseline will be calculated as: 
• 
Percent change from diagnosis (%) = (Change from diagnosis at screening or fist dosing visit / diagnosis value) 
* 100% 
BMI, in kg/m2, will be calculated as follows:  
• 
BMI (kg/ m2) = weight (kg)/ [height (m)2] 
 
13. MEDICATIONS 
All medications will be coded using the World Health Organization (WHO) Drug Global dictionary, version B3 
March 2020. Prior medications are defined as any medication that stopped before the date of first dose of study drug. 
Concomitant medications are defined as any medication that were ongoing or ended at the date of first dose of study 
drug, or any medication that started on or after the date of first dose of study drug. 
Partially or completely missing medication start and stop dates will be handled as described in APPENDIX 1. 


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Prior and concomitant medications will be summarized separately by Anatomical Therapeutic Class (ATC) level 2 
and preferred drug name by treatment arm and overall based on the safety analysis population. A subject having 
more than one medication within the same ATC Level 2 or preferred drug name will be counted only once for that 
ATC Level 2 or preferred drug name. 
All medications (prior and concomitant) will be listed. 
14. EXPOSURE TO STUDY DRUG 
Exposure to the study drug will be summarized by treatment group based on the SAF analysis population for the 
following measures: 
• 
Treatment duration, weeks 
• 
Cumulative drug exposure, mL 
• 
Total number of doses administered, n  
• 
Dose intensity, % 
• 
Any dose modification (dose held, interrupted, reduced, or permanently withdrawn) 
14.1. DERIVATIONS 
Measures of drug exposure will be calculated as follows: 
• 
Treatment duration (weeks) = (Date of last injection of study drug – Date of first injection of study drug + 
30.4167) /7 
• 
Cumulative drug exposure (mL) = the sum of “Volume Administered (mL)” recorded on the Dose eCRF page, 
across all study dosing days 
• 
Total number of doses administered = count of records on the Dose eCRF page where the response to the 
question “Was IP/Placebo administered at this visit” is Yes. 
• 
Dose intensity (%) = (Cumulative drug exposure (mL) / Total planned dose (mL) )*100, where total planned 
dose = (20 mL * 3) 
The dates of first and last AVT001 administration will be taken from the eCRF Dose page. 


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15. COMPLIANCE WITH STUDY DRUG 
Because IP will be administered intravenously to the subjects enrolled in the study in accordance with the protocol, 
compliance will not be assessed. 
16. EFFICACY ENDPOINTS 
Unless otherwise indicated, all efficacy summaries and figures will be presented by treatment group and visit, when 
appropriate, based on the SAF analysis population. 
16.1. PRIMARY EFFICACY 
There are no primary efficacy endpoints in this study. 
16.2. SECONDARY EFFICACY 
The secondary efficacy endpoints analyzed in this SAP are: 
• 
Change from baseline in the area under the curve (AUC) of the stimulated C-peptide levels over the first 4 
hours of a mixed meal tolerance test (MMTT). 
• 
Change from baseline in HbA1c. 
• 
Change from baseline in total, basal, and bolus daily insulin dose. 
• 
Changes from baseline in autoantibody levels. 
16.2.1. 
SECONDARY EFFICACY ENDPOINT VARIABLES AND DERIVATIONS 
16.2.1.1. 
Change from Baseline in the Area under the Curve (AUC) of the stimulated C-
peptide levels over the first 4 hours of a mixed meal tolerance test (MMTT)  
The area under the stimulated C-peptide curve (AUC) over the first 4-hour period of a mixed meal glucose tolerance 
test will be calculated using the trapezoidal rule that is a weighted sum of the C-peptide values over the 240 minutes. 
Missing C-peptide levels at any given timepoint will not be imputed. In the calculation of the AUC when C-peptide 
levels are missing, a line will be drawn from the last timepoint with a non-missing C-peptide to the next timepoint 


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with non-missing C-peptide.  
Change from baseline in C-peptide AUC will be computed for each post-baseline visit where MMTT was a 
scheduled assessment. 
16.2.1.2. 
Change from Baseline in HbA1c 
HbA1c will be measured at the first screening visit and at each dosing and long-term follow-up visit as per the 
schedule of events (refer to protocol Table 3). 
Change from baseline in HbA1c will be computed for each post-baseline visit. 
16.2.1.3. 
Change from Baseline in Total Daily Insulin Dose, Total Daily Basal Insulin Dose, 
and Total Daily Bolus Insulin Dose 
Total daily insulin will be entered by subjects on an insulin log for 3-7 days prior to each study visit starting with 
Study Day 1.  
Change from baseline in total daily insulin dose, total daily basal insulin dose, and total daily bolus insulin dose will 
be computed for each post-baseline visit. 
16.2.1.4. 
Change from Baseline in Autoantibody Levels 
Antibody levels to Glutamic acid decarboxylase (GAD65), Insulinoma associated protein (IA-2), Zinc transporter 8 
(ZnT8), and Insulin associated aAb (IAA) will be measured at the first screening visit and at each dosing and long-
term follow-up visit as per the schedule of event (refer to Protocol Table 3).  
Change from baseline for each antibody type will be computed for each post-baseline visit. 
16.2.2. 
ANALYSIS OF SECONDARY EFFICACY ENDPOINTS 
16.2.2.1. 
Continuous Secondary Efficacy Endpoints 
Observed and change from baseline for C-peptide AUC,  HbA1c, total daily insulin dose, total daily basal insulin 
dose, total daily bolus insulin dose, and each of 4 antibodies will be summarized by treatment group and visit.   
For the analysis of C-peptide AUC, a Mixed-effect Model for Repeated Measurements (MMRM) will be performed 
based on the PD analysis population using the observed-case data.  This model will include the change from baseline 


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variable as the dependent variable, its baseline observed value as covariate, treatment group (active and placebo), 
visit, HbA1c, BMI, total insulin, potency assay, and treatment group-by-visit interaction as fixed effects. A 
backward stepwise variable selection process will be used to remove any non-relevant or highly correlated variables 
before fitting the final model. Restricted Maximum Likelihood (ReML) estimation will be used.  An unstructured 
(UN) covariance structure will be used to model the within-patient error and an adjustment to the degrees of 
freedom will be made using the Kenward-Roger’s approximation.  In the event this model does not converge, the 
heterogeneous Toeplitz, heterogeneous first-order autoregressive (AR(1)), Toeplitz, and AR(1) covariance structures 
will be tested one after the other in the order listed here i.e., the heterogeneous Toeplitz covariance structure will be 
tested first and if this model converges, the other covariance structures will not be tested.  If this model does not 
converge, then the heterogeneous AR(1) will then be tested.  The same process will be repeated for each covariance 
structure one after the other as listed above until the model converges. 
The Least Square (LS) mean estimates will be provided for each treatment group for each visit along with their 
standard error (SE). The difference of LS means between the treatment groups (active vs. placebo) and associated 
SE, two-sided 95% CI, and p-value will also be provided for each visit. 
All remaining secondary endpoints (HbA1c, insulin, and auto-antibody levels) will be summarized univariately by 
generating summaries of the observed values and changes from baseline (both observed and percent changes) by 
study visit. 
 
16.3. EXPLORATORY EFFICACY 
The exploratory efficacy endpoints analyzed in this SAP are: 
• 
Change from baseline in unstimulated C-peptide of a MMTT. 
• 
Change from baseline in the change in the AUC of C-peptide (Delta AUC) over the first 4 hours of a MMTT. 
• 
Change from baseline in the maximum concentration of C-peptide (Cmax) during a MMTT. 
• 
Change from baseline in the maximum C-peptide levels (Delta Cmax) over the first 4 hours of a mixed meal 
tolerance test (MMTT). 
• 
Change from baseline in the total area under the curve (AUC) of the stimulated glucose levels over the first 4 
hours of a mixed meal tolerance test (MMTT). 


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• 
Change from baseline in unstimulated glucose of a MMTT. 
• 
Change from baseline in the change in the AUC of glucose (Delta AUC) over the first 4 hours of a MMTT 
minus the unstimulated glucose AUC prior to MMTT. 
• 
Change from baseline in the maximum concentration of glucose (Cmax) during a MMTT. 
• 
Change from baseline in the maximum change from baseline concentration (Delta Cmax) of the stimulated 
glucose levels over the first 4 hours of a mixed meal tolerance test (MMTT) 
16.3.1. 
EXPLORATORY EFFICACY ENDPOINT VARIABLES AND DERIVATIONS  
16.3.1.1. 
Change from Baseline in unstimluated C-peptide of a MMTT 
Unstimulated C-peptide will be defined as the average of C-peptide at time -10 and 0 minutes of a MMTT. 
16.3.1.2. 
Change from Baseline in the change in AUC of C-peptide (Delta AUC) of a MMTT 
The change in AUC of C-peptide (Delta AUC) will be defined as total AUC C-peptide over the first 4 hours of a 
MMTT minus the unstimulated C-peptide AUC at time -10 and 0 minutes of the same MMTT. 
16.3.1.3. 
Change from Baseline in the maximum concentration of C-peptide (Cmax) of a 
MMTT 
The maximum concentration of C-peptide will be defined as the maximum C-peptide value over the first 4 hours of 
a MMTT. 
16.3.1.4. 
Change from Baseline in the maximum change from baseline concentration (Delta 
Cmax) of the stimulated C-peptide levels of a mixed meal tolerance test (MMTT)  
The maximum change from baseline in C-peptide concentration (Delta Cmax) over the first 4-hour period of a 
mixed meal glucose tolerance test is defined as the maximum concentration of C-Peptide during the MMTT test 
minus the baseline concentration of C-Peptide, which is calculated as the average of C-Peptide at time -10 min and 0 
min of the same MMTT test. Missing C-peptide levels at any given timepoint will not be imputed.  
The maximum delta C-peptide Cmax will be computed for each post-baseline visit where MMTT was a scheduled 
assessment. 


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16.3.1.5. 
Change from Baseline in the AUC of the stimulated glucose of a MMTT  
The area under the stimulated glucose curve (AUC) over the first 4-hour period of a mixed meal glucose tolerance 
test will be calculated the trapezoidal rule as described in Section 16.2.1.  
16.3.1.6. 
Change from Baseline in unstimluated glucse of a MMTT 
Unstimulated glucose will be defined using the approach described in Section 16.3.1.1. 
16.3.1.7. 
Change from Baseline in the change in AUC of glucose (Delta AUC) of a MMTT 
The change in AUC of glucose (Delta AUC) over the first 4-hour period of a MMTT will be defined using the 
approach described in Section 16.3.1.2. 
16.3.1.8. 
Change from Baseline in the maximum concentration of glucose (Cmax) of a 
MMTT 
The maximum glucose concentration (Cmax) over the first 4-hour period of a MMTT will be derived using the 
approach described in Section 16.3.1.3. 
16.3.1.9. 
Change from Baseline in the maximum change from baseline concentration (Delta 
Cmax) of the stimulated glucose levels of a MMTT  
The maximum change from baseline in glucose concentration (Delta Cmax) over the first 4-hour period of a MMTT 
will be derived using the approach described in Section 16.3.1.4.  
16.3.2. 
ANALYSIS OF EXPLORATORY EFFICACY ENDPOINTS 
16.3.2.1. 
Continuous Exploratory Efficacy Endpoints 
Observed and change from baseline for all exploratory endpoints will be summarized by treatment group and visit.  
Univariate summaries of the observed values and changes from baseline (both observed and percent change) will be 
generated by study visit. 


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17. SAFETY ENDPOINTS 
All safety summaries will be presented by treatment group based on the SAF population.  There will be no statistical 
comparisons between the treatment groups for safety data. 
17.1. ADVERSE EVENTS 
• 
Prior adverse events (AEs) are defined as any AE that started or worsened in severity on or after the date of 
signed informed consent but before the first dose of study drug. 
• 
Treatment-emergent AEs (TEAEs) are defined as any AE that starts on or after the first dose of study 
medication. This includes events that start within 30 days following the last dose. 
• 
Post-treatment AEs are defined as adverse events that start more than 30 days following the last dose. 
See APPENDIX 1 for handling of partial dates for AEs.  In the case where it is not possible to define an AE as 
treatment emergent or not, the AE will be classified by the worst case; i.e. treatment emergent.  
AEs will be coded using the MedDRA dictionary, version 22.0. 
An overall summary of number and percentage of subjects within each of the categories described in the sub-
sections below will be provided by treatment group based on the SAF analysis population.  Should a subject 
experience multiple events within a category, the subject will be counted only once for that category. Each adverse 
event summary table will include a sub-table summarizing TEAEs and a sub-table summarizing post-treatment AEs. 
All AEs (prior, TEAE, and post-treatment) will be listed. The listing will include a column to identify potential 
dose-limiting toxicities as collected on the AE CRF page. 
17.1.1. 
ALL TEAES AND POST-TREATMENT AES 
Number and percentage of subjects with at least one TEAE will be presented by SOC and PT.  Should a subject 
experience multiple events within a SOC or PT, the subject will be counted only once for that SOC or PT. Subjects 
with at least one post-treatment AE will be summarized similarly. 
Number and percentage of subjects with at least one TEAE will be broken down further by maximum severity. 
Subjects with at least one post-treatment AE will be summarized similarly. 


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17.1.1.1. 
CTCAE Grading for AEs 
Severity will be classified according to the Common Terminology Criteria for AEs (CTCAE), version 4.03. TEAEs 
or post-treatment AEs starting after the first injection of study drug with a missing CTCAE grade will be classified 
as grade 3. Should a subject experience multiple events within a SOC or PT, only the subject’s worst CTCAE grade 
will be counted for that SOC or PT.  
17.1.1.2. 
Relationship to Study Drug 
Relationship to study drug, as indicated by the Investigator, will be classified as unrelated, unlikely related, possibly 
related, probably related, or definitely related (increasing severity of relationship). A summary of TEAEs related to 
study drug by SOC and PT will be prepared. A sub-table of post-treatment AEs related to study by SOC and PT will 
also be prepared. 
A “Related” AE is defined as a TEAE or a post-treatment AE with a relationship to study drug of possibly related, 
probably related, or definitely related, while a “Non-related” AE is defined as a TEAE or post-treatment AE with a 
relationship to study drug of unrelated or unlikely related.  TEAEs or post-treatment AEs with a missing relationship 
to study drug will be regarded as related to study drug. Should a subject experience multiple events within a SOC or 
PT, only the subject’s worst relationship will be counted for that SOC or PT.  
17.1.2. 
 THE LISTING OF ALL AES WILL INCLUDE A COLUMN FOR RELATEDNESS.ADVERSE 
EVENTS WITH AN OUTCOME OF DEATH 
AEs with an outcome of death are those events which are recorded as “Fatal” on the AE page of the eCRF.  
17.1.3. 
 THE LISTING OF ALL AES WILL INCLUDE A COLUMN FOR OUTCOME.SERIOUS 
ADVERSE EVENTS 
Serious adverse events (SAEs) are those events recorded as “Serious” on the AE page of the eCRF.  A summary of 
serious TEAEs by SOC and PT will be prepared.  A sub-table of post-treatment serious AEs by SOC and PT will 
also be prepared. Should a subject experience multiple events within a SOC or PT, the subject will be counted only 
once for that SOC or PT. 
A listing of all SAEs will be provided 


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17.1.4. 
AES LEADING TO DISCONTINUATION OF STUDY DRUG 
TEAEs leading to permanent discontinuation of study drug are those events recorded as “Drug Permanently 
Discontinued” on the AE pages of the eCRF. A summary of TEAEs leading to permanent discontinuation of study 
drug by SOC and PT will be prepared.  A sub-table of post-treatment AEs leading to discontinuation of study drug 
by SOC and PT will also be prepared. 
A listing of all AEs leading to permanent discontinuation of study drug will be provided. 
17.1.5. 
SEVERE HYPOGLYCAEMIC ADVERSE EVENTS 
Severe hypoglycaemic adverse events will be the AEs coded to the “Hypoglycaemia” Preferred Term (PT). A listing 
of all severe hypoglycaemic events will be provided. A summary table of severe hypoglycaemic adverse events may 
be provided if needed. 
17.1.6. 
LOCAL IV-SITE REACTIONS 
Local IV-site reactions will be the AEs coded to the “Injection site reaction” MedDRA High Level Term (HLT). A 
listing of all IV-site reactions will be provided. A summary table of local IV-site reaction adverse events may be 
provided if needed. 
17.2. DEATHS 
A listing of all deaths will be provided. 
17.3. LABORATORY EVALUATIONS 
A urine pregnancy test will be performed at screening.  Chemistry, hematology, and urinalysis (microalbumin panel) 
will be performed as per the schedule of events (refer to protocol, Table 3).  A list of laboratory parameters to be 
included in the outputs is included in APPENDIX 2.  
Quantitative laboratory parameters reported as “< X”, i.e. below the lower limit of quantification (BLQ) or “> X”, 
i.e. above the upper limit of quantification (ULQ), will be converted to X for the purpose of quantitative summaries, 
but will be presented as recorded, i.e. as “< X” or “> X” in the listings.   
The following summaries will be provided by treatment group based on the SAF analysis population for each 


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chemistry, hematology, and urine microalbumin laboratory parameter: 
• 
Observed and change from baseline in Standard International (SI) units by visit (for quantitative parameters) 
• 
Shift from baseline to the worst post-baseline observed value according to the Common Terminology Criteria 
for Adverse Events (CTCAE) toxicity grades (for quantitative parameters with available CTCAE toxicity 
grades; refer to Section 17.3.11) 
• 
Shifts from baseline to the maximum/minimum post-baseline observed value according to normal range criteria 
(for quantitative parameters without CTCAE toxicity grades; refer to Section 17.3.12) 
All laboratory data will be listed. 
17.3.1. 
CTCAE TOXICITY GRADES 
Quantitative laboratory parameters with available CTCAE toxicity grades will be categorized as follows where 
higher grades representing a more severe toxicity.   
• 
Grade 1 (i.e., mild) 
• 
Grade 2 (i.e., moderate) 
• 
Grade 3 (i.e., severe) 
• 
Grade 4 (i.e., life-threatening) 
• 
Grade 5 (i.e., death) 
Although not defined in the CTCAE toxicity grading system, version 4.03, non-missing laboratory parameter results 
not meeting any of the 5 grades defined in the CTCAE toxicity grading system will be categorized as ‘No Event’ for 
the purpose of the shift from baseline summaries. 
17.3.2. 
LABORATORY NORMAL RANGES 
Quantitative laboratory parameters will be compared with the relevant laboratory normal ranges in SI units and 
categorized as: 
• 
Low: Below the lower limit of the laboratory normal range.  
• 
Normal: Within the laboratory normal range (upper and lower limit included). 


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• 
High: Above the upper limit of the laboratory normal range. 
17.4. VITAL SIGNS 
The following vital sign parameters will be collected for this study as per the schedule of events (refer to protocol, 
Table 3): 
• 
Systolic blood pressure (SBP) (mmHg) 
• 
Diastolic blood pressure (DBP) (mmHg)  
• 
Pulse rate (beats per minute [bpm]) 
• 
Oxygen saturation (%) 
• 
Body temperature (⁰C) 
• 
Weight (kg) 
The following summaries will be provided by treatment group based on the SAF analysis population for each vital 
sign parameter: 
• 
Observed and change from baseline by visit and timepoint 
• 
Number and percentages of subjects with at least one markedly abnormal post-baseline observed value/change 
from baseline (refer to Section 17.4.1) 
All vital sign data will be listed. Vitals signs meeting one of the markedly abnormal criteria in Section 17.4.1 will be 
identified. 
17.4.1. 
VITAL SIGNS MARKEDLY ABNORMAL CRITERIA 
Markedly abnormal vital sign observed values and/or change from baseline will be identified in accordance with the 
following predefined markedly abnormal criteria: 
Variable 
Unit 
Low 
High 
SBP 
mmHg 
≤ 90 mmHg AND  
change from baseline ≤ -20 mmHg 
≥ 180 mmHg AND 
change from baseline ≥ 20 mmHg 


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Variable 
Unit 
Low 
High 
DBP 
mmHg 
≤ 50 mmHg AND 
change from ≤ -15 mmHg 
≥ 105 mmHg AND 
change from baseline ≥ 15 mmHg 
Pulse rate 
bpm 
≤ 50 bpm AND 
change from baseline ≤ -15 bpm 
≥ 120 bpm AND 
change from baseline ≥ 15 bpm 
Oxygen 
saturation 
% 
< 94 % 
Not applicable 
Body 
temperature 
C 
Not applicable 
≥ 38.3 C AND 
change from baseline ≥ 1.1 C 
Weight 
kg 
Percent change from baseline  
≤ -7.0 % 
Percent change from baseline  
> 7.0 % 
 
17.5. ECG EVALUATIONS 
The following electrocardiogram (ECG) parameters will be measured for this study as per the schedule of events 
(refer to protocol, Table 3): 
• 
PR interval (msec) 
• 
RR interval (msec) 
• 
QRS interval (msec) 
• 
QT (uncorrected) interval (msec) 
• 
Overall ECG evaluation (Investigator’s judgment) 
o 
Normal 
o 
Abnormal, not clinically significant (NCS) 
o 
Abnormal, clinically significant (CS) 
The following summaries will be provided by treatment group based on the SAF analysis population for each ECG 


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parameter: 
• 
Observed and change from baseline by visit (for quantitative parameters) 
• 
Number and percentages of subjects with at least one markedly abnormal post-baseline observed value/change 
from baseline (for quantitative parameters; refer to Section 17.5.11) 
• 
Shift from baseline in overall ECG interpretation to the worst post-baseline assessment 
All ECG data will be listed. ECG results meeting one of the markedly abnormal criteria in Section 17.5.1 will be 
identified. 
17.5.1. 
ECG MARKEDLY ABNORMAL CRITERIA 
Markedly abnormal quantitative ECG parameters will be identified in accordance with the following predefined 
markedly abnormal criteria: 
• 
Observed values for QT (uncorrected) interval will be classified as: 
o 
> 450 msec 
o 
> 480 msec 
o 
> 500 msec 
• 
Change from baseline for QT (uncorrected) interval will be classified as: 
o 
>30 msec increase from baseline 
o 
>60 msec increase from baseline 
It is to be noted that the previous categories are not mutually exclusive, but cumulative.  For example, if a subject 
worst post-baseline QT post-baseline observed value is 490 mmHg, then this subject will be reported once under QT 
> 450 msec and once under QT > 480 msec. 
18. DATA NOT SUMMARIZED OR PRESENTED 
Data that will not be summarized or listed are: 
• 
Results of serology tests performed (i.e., HIV, Hepatitis A, B, and C) at screening for the purpose of 


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determining eligibility. 
 
 


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APPENDIX 1. PARTIAL DATE CONVENTIONS 
ALGORITHM FOR PRIOR ADVERSE EVENTS, TREATMENT-EMERGENT  ADVERSE 
EVENTS, AND POST-TREATMENT ADVERSE EVENTS 
START DATE 
END DATE 
ACTION 
Known 
Known/Partial/ 
Missing 
If (AE start date < study drug start date) or AE is a pre-
initial dose event, then PRAE. 
If study drug start date ≤ AE start date and AE start date <= 
last drug date_30 and AE is not a pre-initial dose event, then 
TEAE. 
If study drug start date <= AE start date and AE start date > 
last drug date + 30  then PTAE 
 
 
 
Partial, but known components 
show that AE started before 
study drug start date or AE is a 
pre-initial dose event 
Known/Partial/ 
Missing 
 
PRAE 
 
 
 
Partial and known components 
show that AE started on or after 
study drug start date  
 
Known/Partial/ 
Missing 
Impute missing AE start day as the first of the month or the 
first study drug dose date, whichever is later.  
If (AE start date < study drug start date) or AE is a pre-
initial dose event, then PRAE. 
If study drug start date ≤ AE start date and AE start date <= 
last drug date+30 and AE is not a pre-initial dose event, 
then TEAE. 
If study drug start date <= AE start date and AE start date > 
last drug date+30 then PTAE 
Note: If an AE started prior to first dose of study drug but worsened after treatment started, it will be reported as a 
separate adverse event with a new AE start date. A pre-initial dose event is defined as an AE that is entered in the 
EDC as occurring prior to infusion (Month 0, Day 1). 


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ALGORITHM FOR PRIOR / CONCOMITANT MEDICATIONS 
START DATE 
STOP DATE 
ACTION 
Known 
Known or 
ongoing 
If medication stop date < study drug start date, assign as prior; 
Otherwise, assign as concomitant.  
Partial 
If known components of medication stop date show that medication stopped 
before study drug start date, assign as prior; 
Otherwise, assign as concomitant. 
Missing, not 
ongoing 
Can never be assigned as prior, therefore assign as concomitant. 
 
 
 
Partial 
Known or 
ongoing 
If medication stop date < study drug start date, assign as prior; 
Otherwise, assign as concomitant. 
Partial 
If known components of medication stop date show that medication stopped 
before study drug start date, assign as prior; 
Otherwise, assign as concomitant. 
Missing, not 
ongoing 
Cannot be assigned as prior, therefore assign as concomitant. 
 
 
 
Missing 
Known or 
ongoing 
If medication stop date < study drug start date, assign as prior; 
Otherwise, assign as concomitant. 
Partial 
If known components of medication stop date show that medication stopped 
before study drug start date, assign as prior; 
Otherwise, assign as concomitant. 
Missing, not 
ongoing 
Assign as concomitant. 
 
 
 


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APPENDIX 2. LABORATORY ASSESSMENTS 
Chemistry (SI unit) 
 
• 
Blood urea nitrogen (BUN) (mmol/L) 
• 
Creatinine (µmol/L) 
• 
Calcium (mmol/L) 
• 
Chloride (mmol/L) 
• 
Potassium (mmol/L) 
• 
Sodium (mmol/L) 
• 
Bicarbonate (mmol/L) 
• 
Glucose (mmol/L) 
• 
Alkaline phosphatase (ALP) (U/L) 
 
• 
Alanine transaminase (ALT) (U/L) 
 
• 
Aspartate transaminase (AST) (U/L) 
 
• 
Albumin (g/L) 
• 
Total protein (g/L) 
• 
Total bilirubin (µmol/L) 
 
• 
HbA1c, L/L 
• 
Creatinine Clearance, mL/s 
 
 
 
 
 
 
 
 


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Hematology (SI unit) 
 
• 
Hemoglobin (g/L) 
 
• 
Hematocrit 
 
• 
Red blood cells (RBC) (x10E12/L) 
 
• 
White blood cell (WBC) count total (x10E9/L) 
 
• 
Platelets (x10E9/L) 
 
• 
Absolute neutrophils count (x10E9/L) 
 
• 
Absolute lymphocyte count (x10E9/L) 
 
• 
Absolute monocyte count (x10E9/L) 
 
• 
Absolute eosinophils count (x10E9/L) 
 
• 
Absolute basophils count (x10E9/L) 
 
 
 


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Urine Microalbumin Panel 
 
• 
Urine Microalbumin (mg/dL) 
 
• 
Urine Creatinine (mg/dL) 
 
• 
Microalbumin/Creatinine Ratio (ratio) 
 
 
 
 
 
 
 
 


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STATISTICAL ANALYSIS PLAN 
 
AVT001-T1D-01 
 
A Phase 1 / 2 Double-Blind, Randomized, Placebo 
Controlled Study of Safety, Tolerability and Potential 
Efficacy of AVOTRES Cell-Based Therapy (AVT001) in 
Patients with Type 1 Diabetes 
 
AUTHOR: CARLOS ALEJANDRO DIAZ 
VERSION NUMBER AND DATE: V2.0, 06DEC2022 
 
 
 


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STATISTICAL ANALYSIS PLAN SIGNATURE PAGE 
Statistical Analysis Plan v2.0 (dated 23Nov2022) for protocol AVT001-T1D-01. 
 
Name 
Signature 
Date (DDMmmYYYY) 
Author: 
Carlos Alejandro Diaz 
 
06Dec2022 
 
Position: 
Sr. Biostatistician 
 
Company: 
IQVIA Biotech 
Upon review of this document, the undersigned approves this version of the Statistical Analysis Plan, authorizing 
that the content is acceptable for the reporting of this study. 
 
Name 
Signature 
Date (DDMmmYYYY) 
Approved by: 
Tuochuan Dong 
 
06Dec2022 
 
Position: 
Biostatistician 
 
Company: 
Avotres, Inc. 
 
 


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MODIFICATION HISTORY 
Unique 
Identifier for 
this Version 
Date of the 
Document 
Version 
Author 
Significant Changes from  
Previous Version 
0.1 
22Dec2021 
Mariana Hildesheim 
Not Applicable – First Version 
0.2 
20Jan2022 
Mariana Hildesheim 
Planned analyses revised to condense TLF 
counts 
0.3 
04Mar2022 
Mariana Hildesheim 
Sponsor review comments addressed 
0.4 
16Mar2022 
Mariana Hildesheim 
Sponsor review comments addressed 
1.0 
28Mar2022 
Mariana Hildesheim 
Final version 
1.1 
26Ago2022 
Carlos Alejandro Díaz 
Text updates to include the changes described 
in the memo-to-file (22Apr2022) to amend the 
SAP v1.0, as well as to include the 
‘Supplemental Analysis’ described in section 
4.3, and additional MMRM in section 16.2.2.1. 
1.2 
10Nov2022 
Carlos Alejandro Díaz 
Draft version including Day 360 Analysis 
2.0 
06Dec2022 
Carlos Alejandro Díaz 
Final version sign-off, including Day 360 
Analysis 
 
 
 
 


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TABLE OF CONTENTS 
1. 
INTRODUCTION............................................................................................................... 8 
2. 
STUDY OBJECTIVES AND ENDPOINTS ..................................................................... 8 
2.1. 
Primary Objective and Endpoints .................................................................................................................. 8 
2.2. 
Secondary Objectives and Endpoints ............................................................................................................. 9 
2.3. 
Exploratory Objectives and Endpoints .......................................................................................................... 9 
3. 
STUDY DESIGN ............................................................................................................... 10 
3.1. 
General Description ....................................................................................................................................... 10 
3.2. 
Schedule of Events ......................................................................................................................................... 10 
3.3. 
Changes to Analyses from Protocol.............................................................................................................. 10 
4. 
PLANNED ANALYSES ................................................................................................... 11 
4.1. 
Final analysis after the last subject completes the Month 24 long-term follow-up visit. Data Safety 
Monitoring Board (DSMB) ....................................................................................................................................... 11 
4.2. 
Primary Analysis ........................................................................................................................................... 11 
4.3. 
Supplemental Analysis .................................................................................................................................. 12 
4.4. 
Final Analysis ................................................................................................................................................. 12 
5. 
ANALYSIS POPULATIONS .......................................................................................... 12 
5.1. 
All Screened Subjects Analysis Population ................................................................................................. 12 
5.2. 
Safety Population ........................................................................................................................................... 12 
5.3. 
Pharmacodynamic Population ...................................................................................................................... 13 
5.4. 
Process for Analysis Population Assignment............................................................................................... 13 


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6. 
GENERAL CONSIDERATIONS ................................................................................... 13 
6.1. 
Reference Start Date and Study Day ........................................................................................................... 13 
6.2. 
Baseline ........................................................................................................................................................... 13 
6.3. 
Unscheduled Visits, Retests, and Early Termination Data ........................................................................ 14 
6.4. 
Windowing Conventions ............................................................................................................................... 14 
6.5. 
Common Calculations ................................................................................................................................... 14 
7. 
STATISTICAL CONSIDERATIONS ............................................................................ 14 
7.1. 
Sample Size Calculation ................................................................................................................................ 15 
7.2. 
Missing data ................................................................................................................................................... 15 
7.3. 
Statistical Tests .............................................................................................................................................. 15 
7.4. 
Multiple Comparisons/ Multiplicity ............................................................................................................. 15 
7.5. 
Multicenter Studies ........................................................................................................................................ 16 
7.6. 
Adjustments for Covariates and Factors to be Included in Analyses ....................................................... 16 
7.7. 
Examination of Subgroups ............................................................................................................................ 16 
7.8. 
Software Version ............................................................................................................................................ 16 
8. 
OUTPUT PRESENTATIONS ......................................................................................... 16 
9. 
DISPOSITION AND WITHDRAWALS ........................................................................ 16 
9.1. 
Disposition ...................................................................................................................................................... 16 
9.2. 
Protocol Deviations ........................................................................................................................................ 17 
10. 
DEMOGRAPHIC AND OTHER BASELINE CHARACTERISTICS ....................... 17 
10.1. Derivations ..................................................................................................................................................... 18 
11. 
SURGICAL AND MEDICAL HISTORY ...................................................................... 18 


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12. 
DISEASE HISTORY ........................................................................................................ 18 
12.1. Derivations ..................................................................................................................................................... 19 
13. 
MEDICATIONS ............................................................................................................... 19 
14. 
EXPOSURE TO STUDY DRUG ..................................................................................... 20 
14.1. Derivations ..................................................................................................................................................... 20 
15. 
COMPLIANCE WITH STUDY DRUG ......................................................................... 20 
16. 
EFFICACY ENDPOINTS................................................................................................ 21 
16.1. Primary Efficacy ............................................................................................................................................ 21 
16.2. Secondary Efficacy ........................................................................................................................................ 21 
16.2.1. 
Secondary Efficacy Endpoint Variables and derivations ......................................................................... 21 
16.2.1.1. 
Change from Baseline in the Area under the Curve (AUC) of the stimulated C-peptide levels over 
the first 4 hours of a mixed meal tolerance test (MMTT) .................................................................................... 21 
16.2.1.2. 
Change from Baseline in HbA1c ..................................................................................................... 22 
16.2.1.3. 
Change from Baseline in Total Daily Insulin Dose, Total Daily Basal Insulin Dose, and Total Daily 
Bolus Insulin Dose ............................................................................................................................................... 22 
16.2.1.4. 
Change from Baseline in Autoantibody Levels ............................................................................... 22 
16.2.2. 
Analysis of Secondary Efficacy Endpoints .............................................................................................. 22 
16.2.2.1. 
Continuous Secondary Efficacy Endpoints – C peptide AUC ......................................................... 22 
16.2.2.2. 
Continuous Secondary Efficacy Endpoints – HbA1C ..................................................................... 24 
16.2.2.3. 
Continuous Secondary Efficacy Endpoints – Total Daily Insulin Divided by Weight .................... 24 
16.2.2.4. 
Continuous Secondary Efficacy Endpoints – Antibody Levels to Glutamic Acid Decarboxylase 
(GAD65), Insulinoma Associated Protein (IA-2), Zinc Transporter 8 (ZnT8), and Insulin Associated aAb (IAA)
 
24 
16.3. Exploratory Efficacy ..................................................................................................................................... 25 
16.3.1. 
Exploratory Efficacy Endpoint Variables and Derivations ...................................................................... 26 
16.3.1.1. 
Change from Baseline in unstimluated C-peptide of a MMTT........................................................ 26 
16.3.1.2. 
Change from Baseline in the change in AUC of C-peptide (Delta AUC) of a MMTT .................... 26 
16.3.1.3. 
Change from Baseline in the maximum concentration of C-peptide (Cmax) of a MMTT .............. 26 
16.3.1.4. 
Change from Baseline in the maximum change from baseline concentration (Delta Cmax) of the 
stimulated C-peptide levels of a mixed meal tolerance test (MMTT) ................................................................. 26 
16.3.1.5. 
Change from Baseline in the AUC of the stimulated glucose of a MMTT ...................................... 26 
16.3.1.6. 
Change from Baseline in unstimluated glucose of a MMTT ........................................................... 26 
16.3.1.7. 
Change from Baseline in the change in AUC of glucose (Delta AUC) of a MMTT ....................... 27 
16.3.1.8. 
Change from Baseline in the maximum concentration of glucose (Cmax) of a MMTT .................. 27 
16.3.1.9. 
Change from Baseline in the maximum change from baseline concentration (Delta Cmax) of the 


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stimulated glucose levels of a MMTT ................................................................................................................. 27 
16.3.2. 
Analysis of Exploratory Efficacy Endpoints ........................................................................................... 27 
16.3.2.1. 
Continuous Exploratory Efficacy Endpoints ................................................................................... 27 
17. 
SAFETY ENDPOINTS .................................................................................................... 27 
17.1. Adverse Events ............................................................................................................................................... 27 
17.1.1. 
All TEAEs and Post-Treatment AEs ....................................................................................................... 28 
17.1.1.1. 
CTCAE Grading for AEs ................................................................................................................. 28 
17.1.1.2. 
Relationship to Study Drug .............................................................................................................. 28 
17.1.2. 
AEs with an Outcome of Death ............................................................................................................... 29 
17.1.3. 
Serious Adverse Events ........................................................................................................................... 29 
17.1.4. 
AEs Leading to Discontinuation of Study Drug ...................................................................................... 29 
17.1.5. 
Severe Hypoglycaemic Adverse Events .................................................................................................. 29 
17.1.6. 
Local IV-Site Reactions ........................................................................................................................... 30 
17.2. Deaths ............................................................................................................................................................. 30 
17.3. Laboratory Evaluations ................................................................................................................................ 30 
17.3.1. 
CTCAE Toxicity Grades ......................................................................................................................... 30 
17.3.2. 
Laboratory Normal Ranges ...................................................................................................................... 31 
17.4. Vital Signs ....................................................................................................................................................... 31 
17.4.1. 
Vital Signs Markedly Abnormal Criteria ................................................................................................. 32 
17.5. ECG Evaluations ........................................................................................................................................... 33 
17.5.1. 
ECG Markedly Abnormal Criteria........................................................................................................... 33 
18. 
DATA NOT SUMMARIZED OR PRESENTED .......................................................... 34 
APPENDIX 1. PARTIAL DATE CONVENTIONS AND MISSING DATA 
IMPUTATION 35 
Algorithm for Prior Adverse Events, Treatment-Emergent  Adverse Events, and Post-Treatment Adverse 
Events 35 
Algorithm for Prior / Concomitant Medications .................................................................................................... 36 
Algorithm for Potency Assay .................................................................................................................................... 37 
APPENDIX 2. LABORATORY ASSESSMENTS ................................................................ 38 
 
 


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1. INTRODUCTION 
This statistical analysis plan (SAP) describes the rules and conventions to be used in the presentation and analysis of 
efficacy and safety data for protocol AVT001-T1D-01. It describes the data to be summarized and analyzed, 
including specifics of the statistical analyses to be performed.  
This SAP is based on protocol amendment 3, dated 01Feb2021, inclusive of 2 administrative change letters to 
amendment 3, dated 16Jun2021 and 03Nov2021. The SAP V2.0 amendment was done to include the supplemental 
analysis at Day 360. 
2. STUDY OBJECTIVES AND ENDPOINTS 
2.1. PRIMARY OBJECTIVE AND ENDPOINTS 
Objectives  
Endpoints  
To determine the safety and tolerability profile of 
three doses of AVT001 (as compared to placebo) in 
T1D subjects within-vitro-reversible defects in the 
CD8+ T cell regulatory system. 
Incidence of treatment-emergent adverse events 
(TEAEs). 
Clinically significant changes from baseline in clinical 
laboratory parameters. 
Incidence and severity of local IV-site reactions. 
Changes from baseline in vital signs and 
electrocardiograms. 
Incidence of severe hypoglycaemic events. 
 


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2.2. SECONDARY OBJECTIVES AND ENDPOINTS 
Objectives  
Endpoints  
To determine whether, among T1D subjects with in 
vitro-reversible defects in the CD8+ T cell 
regulatory system, infusion of AVT001 will lead to 
an in vivo reconstitution of this system. 
To determine whether, among T1D subjects with in 
vitro-reversible defects in the CD8+ T cell 
regulatory system, infusion of AVT001 will result 
in amelioration of β cell destruction as assessed by 
stimulated C-peptide. 
 
Assessment of the HLA-E-restricted CD8+ T cell 
regulatory activity (“potency assay”). 
Changes from baseline in the area under the curve 
(AUC) of the stimulated C-peptide levels over the first 
4 hours of a mixed meal tolerance test (MMTT). 
Changes from baseline in HbA1c. 
Change from baseline in insulin usage. 
Changes from baseline in autoantibody levels. 
 
2.3. EXPLORATORY OBJECTIVES AND ENDPOINTS 
Although not specifically defined in the protocol, the following endpoints will be examined as exploratory. 
Objectives  
Endpoints  
To explore the effects of infusion of AVT001 on 
stimulated and unstimulated C-peptide and glucose 
measurements made over the first 4 hours of a 
mixed meal tolerance test (MMTT). 
 
 
Changes from baseline in C-peptide and Glucose 
measured over the first 4 hours of a MMTT: 
• 
Unstimulated C-peptide 
• 
Delta AUC C-peptide 
• 
Cmax C-peptide 
• 
Delta Cmax C-peptide 
• 
Total AUC glucose 
• 
Unstimulated glucose 
• 
Delta AUC glucose 
• 
Cmax glucose 
• 
Delta Cmax glucose 


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3. STUDY DESIGN 
3.1. GENERAL DESCRIPTION 
This study is a phase 1 / 2 double-blind, randomized, placebo-controlled study to evaluate the safety and tolerability 
of AVT001 cell-based therapy, and to assess its potential efficacy for the treatment of type 1 diabetes (T1D). A brief 
overview of the trial design is outlined in Table A. 
Table A: 
Study Design 
3.2. SCHEDULE OF EVENTS 
Schedule of events can be found in Table 3 of the protocol. 
3.3. CHANGES TO ANALYSES FROM PROTOCOL 
Potency assay data will be listed in the final Clinical Study Report (CSR).   
In addition, as an exploratory endpoint, the potency assay data will be evaluated internally by Avotres and will be 
reported separately. 
A supplemental analysis that will occur after the last subject completes the Month 12 study visit (Day 360) has been 
added to SAP v2.0. 


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4. PLANNED ANALYSES 
The following analyses will be performed for this study: 
• 
Primary analysis after the last subject completes the Month 5 study visit (after study unblinding). All 
available data will be analyzed.   
• 
Supplemental analysis after the last subject completes the Month 12 study visit (Day 360). 
4.1. FINAL ANALYSIS AFTER THE LAST SUBJECT COMPLETES THE 
MONTH 24 LONG-TERM FOLLOW-UP VISIT. DATA SAFETY 
MONITORING BOARD (DSMB) 
A DSMB was established to monitor subject safety as well as data quality and adherence. Data reports for review by 
the members were generated from the electronic data capture (EDC) system by the IQVIA Biotech Data 
Management team. The IQVIA Biotech unblinded randomization statistician had a copy of the randomization 
schedule on hand at the closed sessions to provide the DSMB members with unblinded treatment assignments if 
needed. 
Refer to the DSMB Charter for more details about the DSMB composition, responsibilities, and meeting frequency. 
4.2. PRIMARY ANALYSIS 
The primary analysis had taken place for this study after the last subject completes the Month 5 study visit (SAP 
v1.0 was finalized on 28MAR2022 and amended via memo-to-file on 22APR2022). Data used of the primary 
analysis was extracted and locked 29APR2022 04:04, except the potency assay. Unblinding was performed on 
17MAY2022, after the transfer of potency assay data from Avotres had been completed. All analyses described in 
SAP v1.0 and the binding amendment plan were produced and delivered to the sponsor on 10JUN2022. This 
analysis formed the primary basis for the assessment of the study objectives. 
All planned analyses identified in the SAP had been performed by IQVIA Biotech Biostatistics following Sponsor 
Authorization of this SAP, Sponsor authorization of the analysis sets, database lock (DBL), and general study 
unblinding. 


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4.3. SUPPLEMENTAL ANALYSIS 
A supplemental analysis will take place for this study after the last subject completes the Month 12 study visit (Day 
360). The study database will undergo an interim lock at this time. All analyses described in the current document 
will be produced for the supplemental analysis. This analysis will add information to the assessment of the study 
objectives, since data at Day 360 is generally accepted by the T1D medical community as sufficient long-term data 
to evaluate the efficacy and safety of a clinical candidate (AVT001). Moreover, data at Day 360 will facilitate the 
discussion of the development milestones for AVT001. 
All planned analyses identified in the SAP will be performed by IQVIA Biotech Biostatistics following Sponsor 
authorization of this SAP, and database lock (DBL). The study unblinding has already occurred at the time of the 
primary analysis. 
4.4. FINAL ANALYSIS  
The final analysis will take place once the last subject completes the Month 24 long-term follow-up visit. The SAP 
amendment for the final analysis will be updated before the corresponding DBL. 
5. ANALYSIS POPULATIONS 
5.1. ALL SCREENED SUBJECTS ANALYSIS POPULATION 
The all screened subjects (SCRN) analysis population will contain all subjects who provide informed consent for 
this study. 
5.2. SAFETY POPULATION 
The safety (SAF) population is defined as all subjects who receive at least one dose of study medication. If there is 
any doubt whether a subject was treated or not, he/she will be assumed treated for the purposes of analysis. 
For analyses and displays based on SAF, subjects will be classified according to the actual treatment received. 


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5.3. PHARMACODYNAMIC POPULATION 
The pharmacodynamic (PD) population is defined as all subjects in the safety population with at least one post-
baseline assessment for the evaluation of the CD8+ T-cell reg system biomarker. If there is any doubt whether a 
subject was treated or not, he/she will be assumed treated for the purposes of analysis.  
For analyses and displays based on PD, subjects will be classified according to the actual treatment received. 
5.4. PROCESS FOR ANALYSIS POPULATION ASSIGNMENT 
For the PD population, the identification and agreement of protocol deviations or events which affect PD results will 
be performed between the biostatistician, and the sponsor, prior to DBL, with sponsor authorization of any excluded 
subjects or their data. 
6. GENERAL CONSIDERATIONS 
6.1. REFERENCE START DATE AND STUDY DAY 
Study Day will be calculated from the reference start date and will be used to show start/stop day of assessments and 
events. Reference start date is defined as the day of the first injection of study drug i.e., Day 1. 
Study Day will be computed as follows: 
• 
Study Day = (Date of event –Date of first injection of study drug) + 1 if the date of the event is on or after the 
date of the first injection of study drug 
• 
Study Day = (Date of event – Date of first injection of study drug) if the date of the event is prior to the date of 
the first injection of study drug 
In the situation where the event date is partial or missing, Study Day and any corresponding durations will appear 
partial or missing in the listings. 
6.2. BASELINE 
Unless otherwise specified, baseline is defined as the last non-missing measurement taken prior to the first injection 
of study drug (including unscheduled assessments). In the case where the last non-missing measurement and the date 


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and time of the first injection of study drug coincide, that measurement will be considered pre-baseline, but adverse 
events (AEs) and medications commencing on the date and time of the first injection of study drug will be 
considered post-baseline. 
6.3. UNSCHEDULED VISITS, RETESTS, AND EARLY TERMINATION 
DATA 
For by-visit summaries, data recorded at the nominal visit will be presented. That is, unscheduled, retest (same visit 
number assigned), and early termination measurements will not be included in by-visit summaries but might 
contribute to the baseline timepoint and/or maximum value, where required (e.g. shift table).  
Listings will include scheduled, unscheduled, retest and early discontinuation data. 
6.4. WINDOWING CONVENTIONS  
No visit windowing for analysis purposes will be performed for this study.  
6.5. COMMON CALCULATIONS 
Change from baseline will be calculated as: 
• 
Change from baseline = Test value at baseline or post-baseline visit – Baseline value 
Percent change from baseline will be calculated as: 
• 
Percent change from baseline (%) = (Change from baseline at baseline or post-baseline visit / Baseline value) * 
100% 
For select parameters where data is available, change and percent change will also be calculated from diagnosis. 
7. STATISTICAL CONSIDERATIONS 
For continuous data, descriptive statistics (i.e., n [number of subjects with available data], mean, standard deviation 
[SD], median, 25th and 75th percentiles, and minimum, and maximum values) will be presented by treatment group 
and visit, when applicable. 


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For categorical data, the number and percentages of subjects in each category will be presented by treatment group 
and visit, when applicable 
7.1. SAMPLE SIZE CALCULATION 
The sample size of 14 subjects treated with AVT001 will provide at least 80% power to observe at least one 
occurrence of treatment-emergent adverse events with a true underlying incidence of 12%. The sample size provides 
the same power to detect at least one occurrence of local tolerability events relating to the i.v. administration of 
study drug. 
To account for an estimated 10% potential dropout prior to the completion of the Month 5 safety/efficacy 
assessment post-first dose, a total sample size of approximately 24 subjects will be randomized, in a 2:1 ratio of 
AVT001:Placebo. 
7.2. MISSING DATA 
Missing efficacy data will not be imputed, except for missing potency assay on Visit 3/Day 1, as described in 
APPENDIX 1.  
Missing safety data will not be imputed, except for missing AE severity and relationship data (refer to Section 
17.1.1). Partial or completely missing AE and medication dates will be handled as described in APPENDIX 1. 
7.3. STATISTICAL TESTS 
All statistical tests will be conducted at the two-sided 5% significant level, unless otherwise specified in the 
description of the analyses.  Confidence Intervals (CIs) will be two-sided with 95% coverage. 
7.4. MULTIPLE COMPARISONS/ MULTIPLICITY 
No adjustment for multiple comparisons and multiplicity will be performed.  Only nominal p-values will be 
provided for the efficacy endpoints. 
No statistical testing will be performed for the safety endpoints. 


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7.5. MULTICENTER STUDIES 
This study will be conducted by one investigator at one center. Pooling of data for analysis is not applicable.  
7.6. ADJUSTMENTS FOR COVARIATES AND FACTORS TO BE 
INCLUDED IN ANALYSES 
Mixed-effect Model for Repeated Measurements (MMRM) analyses C-peptide total AUC will be adjusted for 
treatment group, visit, HbA1c, BMI, total daily insulin divided by weight, potency assay, and treatment group-by-
visit interaction as fixed effects. See section 16.2 for further details. 
7.7. EXAMINATION OF SUBGROUPS 
No subgroup analyses are planned.  
7.8. SOFTWARE VERSION 
All analyses will be conducted using SAS version 9.4.  
8. OUTPUT PRESENTATIONS 
The presentation of data in outputs will follow IQVIA Biotech Data Display Standards. Refer to the TFL shells 
document for additional information. 
9. DISPOSITION AND WITHDRAWALS 
All subjects who provide informed consent will be accounted for in this study.  
9.1. DISPOSITION 
Number of subjects screened will be presented overall for the SCRN analysis population.  Number and percentage 
of subjects with screen failure and reason for screen failure will also be presented overall based on the SCRN 
analysis population.  Number of subjects randomized will be presented overall and by treatment group for the SCRN 


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analysis population. 
Number and percentages of subjects treated, who completed/discontinued early from treatment (including reason for 
withdrawal), and who completed/discontinued early from the study (including reason for withdrawal) will be 
provided overall and by treatment group based on the SAF analysis population. 
A listing showing inclusion and exclusion of each subject from each analysis population, including reason for 
exclusion, will be provided. 
9.2. PROTOCOL DEVIATIONS 
A listing of protocol deviations identified by the study team (important or not) will be provided. 
10. DEMOGRAPHIC AND OTHER BASELINE CHARACTERISTICS 
The following demographic and other baseline characteristics collected during the screening visit will be reported 
for this study: 
• 
Age (years) – calculated relative to date of consent 
• 
Age groups 
• 
Sex 
• 
Childbearing potential (for female subjects only) 
• 
Race 
• 
Ethnicity 
• 
Weight (kg) 
• 
Height (cm) 
• 
Body Mass Index (BMI) (kg/m2) 
Continuous demographic and other baseline characteristics will be summarized using descriptive statistics overall 
and by treatment group based on the SAF analysis population.  For categorical demographic and other baseline 
characteristics, number and percentage of subjects in each category will be provided overall and by treatment group 
based on the SAF analysis population.  No statistical testing will be carried out for demographic or other baseline 


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characteristics. 
10.1. DERIVATIONS 
BMI, in kg/m2, will be calculated as follows:  
• 
BMI (kg/ m2) = weight (kg)/ [height (m)2] 
11. SURGICAL AND MEDICAL HISTORY 
Surgical and medical history are defined as any surgeries that happened before the first injection of study drug and 
any medical conditions/diseases that started and stopped before the first injection of study drug.  
Surgical and medical history will be coded using the Medical Dictionary for Regulatory Activities (MedDRA), 
version 22.0, and will be summarized by System Organ Class (SOC) and Preferred Term (PT) overall and by 
treatment group based on the SAF analysis population.  A subject having more than one surgery/medical 
condition/disease within the same SOC/PT will be counted only once for that SOC or PT. 
All surgical and medical history will be listed. 
12. DISEASE HISTORY 
The following disease history characteristics will be summarized overall and by treatment group based on the SAF 
analysis population: 
• 
Time since diagnosis (days) – calculated relative to date of first infusion of study drug 
• 
Use of insulin pump (yes/no) 
• 
HbA1c at diagnosis, screening, and baseline (%) 
• 
Change and percent change in HbA1c from diagnosis to baseline (%) 
• 
Weight at diagnosis, screening, and baseline (kg) 
• 
Change and percent change in weight from diagnosis to baseline (kg) 
• 
BMI at diagnosis, screening, and baseline (kg/m2) 


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• 
Change and percent change in BMI from diagnosis to baseline (kg/m2) 
All disease history characteristics will be listed. 
12.1. DERIVATIONS 
‘Time since’ disease history characteristics, in days, will be calculated as follows: 
• 
Time since diagnosis (days) = (Date of first infusion of study drug - Date of diagnosis) 
Change from diagnosis will be calculated as follows: 
• 
Change from diagnosis = Test value at screening or first dosing visit – Diagnosis value 
Percent change from baseline will be calculated as: 
• 
Percent change from diagnosis (%) = (Change from diagnosis at screening or fist dosing visit / diagnosis value) 
* 100% 
BMI, in kg/m2, will be calculated as follows:  
• 
BMI (kg/ m2) = weight (kg)/ [height (m)2] 
13. MEDICATIONS 
All medications will be coded using the World Health Organization (WHO) Drug Global dictionary, version B3 
March 2020. Prior medications are defined as any medication that stopped before the date of first dose of study drug. 
Concomitant medications are defined as any medication that were ongoing or ended at the date of first dose of study 
drug, or any medication that started on or after the date of first dose of study drug. 
Partially or completely missing medication start and stop dates will be handled as described in APPENDIX 1. 
Prior and concomitant medications will be summarized separately by Anatomical Therapeutic Class (ATC) level 2 
and preferred drug name by treatment arm and overall based on the safety analysis population. A subject having 
more than one medication within the same ATC Level 2 or preferred drug name will be counted only once for that 
ATC Level 2 or preferred drug name. 
All medications (prior and concomitant) will be listed. 


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14. EXPOSURE TO STUDY DRUG 
Exposure to the study drug will be summarized by treatment group based on the SAF analysis population for the 
following measures: 
• 
Treatment duration, weeks 
• 
Cumulative drug exposure, mL 
• 
Total number of doses administered, n  
• 
Dose intensity, % 
• 
Any dose modification (dose held, interrupted, reduced, or permanently withdrawn) 
14.1. DERIVATIONS 
Measures of drug exposure will be calculated as follows: 
• 
Treatment duration (weeks) = (Date of last injection of study drug – Date of first injection of study drug + 
30.4167) /7 
• 
Cumulative drug exposure (mL) = the sum of “Volume Administered (mL)” recorded on the Dose eCRF page, 
across all study dosing days 
• 
Total number of doses administered = count of records on the Dose eCRF page where the response to the 
question “Was IP/Placebo administered at this visit” is Yes. 
• 
Dose intensity (%) = (Cumulative drug exposure (mL) / Total planned dose (mL))*100, where total planned 
dose = (20 mL * 3) 
The dates of first and last AVT001 administration will be taken from the eCRF Dose page. 
15. COMPLIANCE WITH STUDY DRUG 
Because IP will be administered intravenously to the subjects enrolled in the study in accordance with the protocol, 
compliance will not be assessed. 


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16. EFFICACY ENDPOINTS 
Unless otherwise indicated, all efficacy summaries and figures will be presented by treatment group and visit, when 
appropriate, based on the SAF analysis population. 
16.1. PRIMARY EFFICACY 
There are no primary efficacy endpoints in this study. 
16.2. SECONDARY EFFICACY 
The secondary efficacy endpoints analyzed in this SAP are: 
• 
Change from baseline in the area under the curve (AUC) of the stimulated C-peptide levels over the first 4 
hours of a mixed meal tolerance test (MMTT). 
• 
Change from baseline in HbA1c. 
• 
Change from baseline in total, basal, and bolus daily insulin dose. 
• 
Changes from baseline in autoantibody levels. 
16.2.1. 
SECONDARY EFFICACY ENDPOINT VARIABLES AND DERIVATIONS 
16.2.1.1. 
Change from Baseline in the Area under the Curve (AUC) of the stimulated C-
peptide levels over the first 4 hours of a mixed meal tolerance test (MMTT)  
The area under the stimulated C-peptide curve (AUC) over the first 4-hour period of a mixed meal glucose tolerance 
test will be calculated using the trapezoidal rule that is a weighted sum of the C-peptide values over the 240 minutes. 
Missing C-peptide levels at any given timepoint will not be imputed. In the calculation of the AUC when C-peptide 
levels are missing, a line will be drawn from the last timepoint with a non-missing C-peptide to the next timepoint 
with non-missing C-peptide.  
Change from baseline in C-peptide AUC will be computed for baseline and for each post-baseline visit where 
MMTT was a scheduled assessment. 


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16.2.1.2. 
Change from Baseline in HbA1c 
HbA1c will be measured at the first screening visit and at each dosing and long-term follow-up visit as per the 
schedule of events (refer to protocol Table 3). 
Change from baseline in HbA1c will be computed for baseline and for each post-baseline visit. 
16.2.1.3. 
Change from Baseline in Total Daily Insulin Dose, Total Daily Basal Insulin Dose, 
and Total Daily Bolus Insulin Dose 
Total daily insulin will be entered by subjects on an insulin log for 3-7 days prior to each study visit starting with 
Study Day 1.  Change from baseline in total daily insulin dose, total daily basal insulin dose, and total daily bolus 
insulin dose will be computed for baseline and for each post-baseline visit. 
Because insulin is heavily dependent on weight at the same visit, the following measures will also be computed for 
baseline and for each post-baseline visit: change from baseline in total daily insulin dose divided by weight, total 
daily basal insulin dose divided by weight, and total daily bolus insulin dose divided by weight. This additional 
summary is included per the 22Apr2022 memo-to-file to amend the SAP v1.0. 
16.2.1.4. 
Change from Baseline in Autoantibody Levels 
Antibody levels to glutamic acid decarboxylase (GAD65), insulinoma associated protein (IA-2), zinc transporter 8 
(ZnT8), and insulin associated aAb (IAA) will be measured at the first screening visit and at each dosing and long-
term follow-up visit as per the schedule of event (refer to Protocol Table 3).  
Change from baseline for each antibody type will be computed for baseline and for each post-baseline visit. 
16.2.2. 
ANALYSIS OF SECONDARY EFFICACY ENDPOINTS 
Observed value and change from baseline for C-peptide AUC, HbA1c, total daily insulin dose, total daily basal 
insulin dose, total daily bolus insulin dose, and each of 4 antibodies will be summarized by treatment group and 
visit.   
16.2.2.1. 
Continuous Secondary Efficacy Endpoints – C peptide AUC 
For the primary analysis of C-peptide AUC, a Mixed-effect Model for Repeated Measurements (MMRM) will be 
performed based on the PD analysis population using the observed-case data.  This model will include the change 


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from baseline variable as the dependent variable, its baseline observed value, age, and time from diagnosis to the 
first dose as covariates, treatment group (active and placebo), visit, HbA1c, BMI, total daily insulin divided by 
weight, potency assay, and treatment group-by-visit interaction as fixed effects. The fixed effects and covariates 
specified in SAP v1.0 were revised per the 22APR2022 memo-to-file to amend the SAP v1.0. 
A backward stepwise variable selection process will be used to remove any non-relevant or highly correlated 
variables before fitting the final model. Restricted Maximum Likelihood (ReML) estimation will be used.  An 
unstructured (UN) covariance structure will be used to model the within-patient error and an adjustment to the 
degrees of freedom will be made using the Kenward-Roger’s approximation.  In the event this model does not 
converge, the heterogeneous Toeplitz, heterogeneous first-order autoregressive (AR(1)), Toeplitz, and AR(1) 
covariance structures will be tested one after the other in the order listed here i.e., the heterogeneous Toeplitz 
covariance structure will be tested first and if this model converges, the other covariance structures will not be 
tested.  If this model does not converge, then the heterogeneous AR(1) will then be tested.  The same process will be 
repeated for each covariance structure one after the other as listed above until the model converges. 
The Least Square (LS) mean estimates will be provided for each treatment group for each visit along with their 
standard error (SE). The difference of LS means between the treatment groups (active vs. placebo) and associated 
SE, two-sided 95% CI for each visit, and p-value will also be provided only for Visits at Day 150 and Day 360. 
In addition, the following MMRMs for C-peptide AUC will be analyzed as exploratory analyses: 
• 
MMRM, based on the data locked for the primary analysis when the last subject completes the Month 5 
study visit (Day 150), with the data at Visit 3/Day 1 included. A separate backward stepwise variable 
selection process will be performed. 
• 
MMRM, based on the data locked for the primary analysis when the last subject completes the Month 5 
study visit (Day 150), with the data at Visit 3/Day 1 excluded. A separate backward stepwise variable 
selection process will be performed. 
• 
MMRM, based on the data locked for the supplemental analysis when the last subject completes the Month 
12 study visit (Day 360), with the data at Visit 3/Day 1 excluded. A separate backward stepwise variable 
selection process will be performed. 
• 
MMRM, based on the data locked for the supplemental analysis when the last subject completes the Month 
12 study visit (Day 360), with the data at Visit 3/Day 1 excluded. The full model will be fitted directly 
without any variable selection process. The primary analysis performed in June 2022 for Day 150 reported 
results from the full model without any variable selection process, and with the data at Visit 3/Day 1 
excluded. 


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16.2.2.2. 
Continuous Secondary Efficacy Endpoints – HbA1C 
For the analysis of HbA1C, a Mixed-effect Model for Repeated Measurements (MMRM) will be performed based 
on the PD analysis population using the observed-case data.  This model will include the change from baseline 
variable as the dependent variable, its baseline observed value, age, and time from diagnosis to the first dose as 
covariates, treatment group (active and placebo), visit, C-peptide AUC, BMI, total daily insulin divided by weight, 
potency assay, and treatment group-by-visit interaction as fixed effects.  
The MMRM modeling and selection procedures described in section 6.2.2.1 will be used to select an appropriate 
model for this endpoint. 
The Least Square (LS) mean estimates will be provided for each treatment group for each visit along with their 
standard error (SE). The difference of LS means between the treatment groups (active vs. placebo) and associated 
SE, two-sided 95% CI for each visit, and p-value will also be provided only for Visits at Day 150 and Day 360. 
16.2.2.3. 
Continuous Secondary Efficacy Endpoints – Total Daily Insulin Divided by Weight 
For the analysis of total daily insulin divided by weight, a Mixed-effect Model for Repeated Measurements 
(MMRM) will be performed based on the PD analysis population using the observed-case data.  This model will 
include the change from baseline variable as the dependent variable, its baseline observed value, age, and time from 
diagnosis to the first dose as covariates, treatment group (active and placebo), visit, C-peptide AUC, BMI, HbA1C, 
potency assay, and treatment group-by-visit interaction as fixed effects.  
The MMRM modeling and selection procedures described in section 6.2.2.1 will be used to select an appropriate 
model for this endpoint. 
The Least Square (LS) mean estimates will be provided for each treatment group for each visit along with their 
standard error (SE). The difference of LS means between the treatment groups (active vs. placebo) and associated 
SE, two-sided 95% CI for each visit, and p-value will also be provided only for Visits at Day 150 and Day 360. 
16.2.2.4. 
Continuous Secondary Efficacy Endpoints – Antibody Levels to Glutamic Acid 
Decarboxylase (GAD65), Insulinoma Associated Protein (IA-2), Zinc Transporter 8 (ZnT8), and 
Insulin Associated aAb (IAA) 
For the analysis of each of the antibody levels to glutamic acid decarboxylase (GAD65), insulinoma associated 
protein (IA-2), zinc transporter 8 (ZnT8), and insulin associated aAb (IAA), a Mixed-effect Model for Repeated 
Measurements (MMRM) will be performed based on the PD analysis population using the observed-case data. This 


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model will include the change from baseline variable as the dependent variable, its baseline observed value, age, and 
time from diagnosis to the first dose as covariates, treatment group (active and placebo), visit, C-peptide AUC, 
HbA1C, BMI, total daily insulin divided by weight, potency assay, and treatment group-by-visit interaction as fixed 
effects.  
The MMRM modeling and selection procedures described in section 6.2.2.1 will be used to select an appropriate 
model for these endpoints. 
The Least Square (LS) mean estimates will be provided for each treatment group for each visit along with their 
standard error (SE). The difference of LS means between the treatment groups (active vs. placebo) and associated 
SE, two-sided 95% CI for each visit, and p-value will also be provided only for Visits at Day 150 and Day 360. 
 
16.3. EXPLORATORY EFFICACY 
The exploratory efficacy endpoints analyzed in this SAP are: 
• 
Change from baseline in unstimulated C-peptide of a MMTT. 
• 
Change from baseline in the change in the AUC of C-peptide (Delta AUC) over the first 4 hours of a MMTT. 
• 
Change from baseline in the maximum concentration of C-peptide (Cmax) during a MMTT. 
• 
Change from baseline in the maximum C-peptide levels (Delta Cmax) over the first 4 hours of a mixed meal 
tolerance test (MMTT). 
• 
Change from baseline in the total area under the curve (AUC) of the stimulated glucose levels over the first 4 
hours of a mixed meal tolerance test (MMTT). 
• 
Change from baseline in unstimulated glucose of a MMTT. 
• 
Change from baseline in the change in the AUC of glucose (Delta AUC) over the first 4 hours of a MMTT 
minus the unstimulated glucose AUC prior to MMTT. 
• 
Change from baseline in the maximum concentration of glucose (Cmax) during a MMTT. 
• 
Change from baseline in the maximum change from baseline concentration (Delta Cmax) of the stimulated 
glucose levels over the first 4 hours of a mixed meal tolerance test (MMTT) 


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16.3.1. 
EXPLORATORY EFFICACY ENDPOINT VARIABLES AND DERIVATIONS  
16.3.1.1. 
Change from Baseline in unstimluated C-peptide of a MMTT 
Unstimulated C-peptide will be defined as the average of C-peptide at time -10 and 0 minutes of a MMTT. 
16.3.1.2. 
Change from Baseline in the change in AUC of C-peptide (Delta AUC) of a MMTT 
The change in AUC of C-peptide (Delta AUC) will be defined as total AUC C-peptide over the first 4 hours of a 
MMTT minus the unstimulated C-peptide AUC at time -10 and 0 minutes of the same MMTT. 
16.3.1.3. 
Change from Baseline in the maximum concentration of C-peptide (Cmax) of a 
MMTT 
The maximum concentration of C-peptide will be defined as the maximum C-peptide value over the first 4 hours of 
a MMTT. 
16.3.1.4. 
Change from Baseline in the maximum change from baseline concentration (Delta 
Cmax) of the stimulated C-peptide levels of a mixed meal tolerance test (MMTT)  
The maximum change from baseline in C-peptide concentration (Delta Cmax) over the first 4-hour period of a 
mixed meal glucose tolerance test is defined as the maximum concentration of C-Peptide during the MMTT test 
minus the baseline concentration of C-Peptide, which is calculated as the average of C-Peptide at time -10 min and 0 
min of the same MMTT test. Missing C-peptide levels at any given timepoint will not be imputed.  
The maximum delta C-peptide Cmax will be computed for baseline and for each post-baseline visit where MMTT 
was a scheduled assessment. 
16.3.1.5. 
Change from Baseline in the AUC of the stimulated glucose of a MMTT  
The area under the stimulated glucose curve (AUC) over the first 4-hour period of a mixed meal glucose tolerance 
test will be calculated the trapezoidal rule as described in Section 16.2.1.  
16.3.1.6. 
Change from Baseline in unstimluated glucose of a MMTT 
Unstimulated glucose will be defined using the approach described in Section 16.3.1.1. 


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16.3.1.7. 
Change from Baseline in the change in AUC of glucose (Delta AUC) of a MMTT 
The change in AUC of glucose (Delta AUC) over the first 4-hour period of a MMTT will be defined using the 
approach described in Section 16.3.1.2. 
16.3.1.8. 
Change from Baseline in the maximum concentration of glucose (Cmax) of a 
MMTT 
The maximum glucose concentration (Cmax) over the first 4-hour period of a MMTT will be derived using the 
approach described in Section 16.3.1.3. 
16.3.1.9. 
Change from Baseline in the maximum change from baseline concentration (Delta 
Cmax) of the stimulated glucose levels of a MMTT  
The maximum change from baseline in glucose concentration (Delta Cmax) over the first 4-hour period of a MMTT 
will be derived using the approach described in Section 16.3.1.4.  
16.3.2. 
ANALYSIS OF EXPLORATORY EFFICACY ENDPOINTS 
16.3.2.1. 
Continuous Exploratory Efficacy Endpoints 
Observed and change from baseline for all exploratory endpoints will be summarized by treatment group and visit.  
Univariate summaries of the observed values and changes from baseline (both observed and percent change) will be 
generated by study visit. 
17. SAFETY ENDPOINTS 
All safety summaries will be presented by treatment group based on the SAF population.  There will be no statistical 
comparisons between the treatment groups for safety data. 
17.1. ADVERSE EVENTS 
• 
Prior adverse events (AEs) are defined as any AE that started or worsened in severity on or after the date of 
signed informed consent but before the first dose of study drug. 
• 
Treatment-emergent AEs (TEAEs) are defined as any AE that starts on or after the first dose of study 


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medication. This includes events that start within 30 days following the last dose. 
• 
Post-treatment AEs are defined as adverse events that start more than 30 days following the last dose. 
See APPENDIX 1 for handling of partial dates for AEs.  In the case where it is not possible to define an AE as 
treatment emergent or not, the AE will be classified by the worst case; i.e. treatment emergent.  
AEs will be coded using the MedDRA dictionary, version 22.0. 
An overall summary of number and percentage of subjects within each of the categories described in the sub-
sections below will be provided by treatment group based on the SAF analysis population.  Should a subject 
experience multiple events within a category, the subject will be counted only once for that category. Each adverse 
event summary table will include a sub-table summarizing TEAEs and a sub-table summarizing post-treatment AEs. 
All AEs (prior, TEAE, and post-treatment) will be listed. The listing will include a column for relatedness (see 
section 17.1.1.2), a column for outcome, and a column to identify potential dose-limiting toxicities as collected on 
the AE CRF page.  Listings of subsets of AEs will include these columns as well. 
17.1.1. 
ALL TEAES AND POST-TREATMENT AES 
Number and percentage of subjects with at least one TEAE will be presented by SOC and PT.  Should a subject 
experience multiple events within a SOC or PT, the subject will be counted only once for that SOC or PT. Subjects 
with at least one post-treatment AE will be summarized similarly. 
Number and percentage of subjects with at least one TEAE will be broken down further by maximum severity. 
Subjects with at least one post-treatment AE will be summarized similarly. 
17.1.1.1. 
CTCAE Grading for AEs 
Severity will be classified according to the Common Terminology Criteria for AEs (CTCAE), version 4.03. TEAEs 
or post-treatment AEs starting after the first injection of study drug with a missing CTCAE grade will be classified 
as grade 3. Should a subject experience multiple events within a SOC or PT, only the subject’s worst CTCAE grade 
will be counted for that SOC or PT.  
17.1.1.2. 
Relationship to Study Drug 
Relationship to study drug, as indicated by the Investigator, will be classified as unrelated, unlikely related, possibly 
related, probably related, or definitely related (increasing severity of relationship). A summary of TEAEs related to 


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study drug by SOC and PT will be prepared. A sub-table of post-treatment AEs related to study by SOC and PT will 
also be prepared. 
A “Related” AE is defined as a TEAE or a post-treatment AE with a relationship to study drug of possibly related, 
probably related, or definitely related, while a “Non-related” AE is defined as a TEAE or post-treatment AE with a 
relationship to study drug of unrelated or unlikely related.  TEAEs or post-treatment AEs with a missing relationship 
to study drug will be regarded as related to study drug. Should a subject experience multiple events within a SOC or 
PT, only the subject’s worst relationship will be counted for that SOC or PT.  
17.1.2. 
AES WITH AN OUTCOME OF DEATH 
AEs with an outcome of death are those events which are recorded as “Fatal” on the AE page of the eCRF.  
17.1.3. 
SERIOUS ADVERSE EVENTS 
Serious Adverse Events (SAEs) are those events recorded as “Serious” on the AE page of the eCRF.  A summary of 
serious TEAEs by SOC and PT will be prepared.  A sub-table of post-treatment serious AEs by SOC and PT will 
also be prepared. Should a subject experience multiple events within a SOC or PT, the subject will be counted only 
once for that SOC or PT. 
A listing of all SAEs will be provided. 
17.1.4. 
AES LEADING TO DISCONTINUATION OF STUDY DRUG 
TEAEs leading to permanent discontinuation of study drug are those events recorded as “Drug Permanently 
Discontinued” on the AE pages of the eCRF. A summary of TEAEs leading to permanent discontinuation of study 
drug by SOC and PT will be prepared.  A sub-table of post-treatment AEs leading to discontinuation of study drug 
by SOC and PT will also be prepared. 
A listing of all AEs leading to permanent discontinuation of study drug will be provided. 
17.1.5. 
SEVERE HYPOGLYCAEMIC ADVERSE EVENTS 
Severe hypoglycaemic adverse events will be the AEs coded to the “Hypoglycaemia” Preferred Term (PT). A listing 
of all severe hypoglycaemic events will be provided. A summary table of severe hypoglycaemic adverse events may 
be provided if needed. 


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17.1.6. 
LOCAL IV-SITE REACTIONS 
Local IV-site reactions will be the AEs coded to the “Injection site reaction” MedDRA High Level Term (HLT). A 
listing of all IV-site reactions will be provided. A summary table of local IV-site reaction adverse events may be 
provided if needed. 
17.2. DEATHS 
A listing of all deaths will be provided. 
17.3. LABORATORY EVALUATIONS 
A urine pregnancy test will be performed at screening.  Chemistry, hematology, and urinalysis (microalbumin panel) 
will be performed as per the schedule of events (refer to protocol, Table 3).  A list of laboratory parameters to be 
included in the outputs is included in APPENDIX 2.  
Quantitative laboratory parameters reported as “< X”, i.e. below the lower limit of quantification (BLQ) or “> X”, 
i.e. above the upper limit of quantification (ULQ), will be converted to X for the purpose of quantitative summaries, 
but will be presented as recorded, i.e. as “< X” or “> X” in the listings.   
The following summaries will be provided by treatment group based on the SAF analysis population for each 
chemistry, hematology, and urine microalbumin laboratory parameter: 
• 
Observed and change from baseline in Standard International (SI) units by visit (for quantitative parameters) 
• 
Shift from baseline to the worst post-baseline observed value according to the Common Terminology Criteria 
for Adverse Events (CTCAE) toxicity grades (for quantitative parameters with available CTCAE toxicity 
grades; refer to Section 17.3.11) 
• 
Shifts from baseline to the maximum/minimum post-baseline observed value according to normal range criteria 
(for quantitative parameters without CTCAE toxicity grades; refer to Section 17.3.12) 
All laboratory data will be listed. 
17.3.1. 
CTCAE TOXICITY GRADES 
Quantitative laboratory parameters with available CTCAE toxicity grades will be categorized as follows where 


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higher grades representing a more severe toxicity.   
• 
Grade 1 (i.e., mild) 
• 
Grade 2 (i.e., moderate) 
• 
Grade 3 (i.e., severe) 
• 
Grade 4 (i.e., life-threatening) 
• 
Grade 5 (i.e., death) 
Although not defined in the CTCAE toxicity grading system, version 4.03, non-missing laboratory parameter results 
not meeting any of the 5 grades defined in the CTCAE toxicity grading system will be categorized as ‘No Event’ for 
the purpose of the shift from baseline summaries. 
17.3.2. 
LABORATORY NORMAL RANGES 
Quantitative laboratory parameters will be compared with the relevant laboratory normal ranges in SI units and 
categorized as: 
• 
Low: Below the lower limit of the laboratory normal range.  
• 
Normal: Within the laboratory normal range (upper and lower limit included). 
• 
High: Above the upper limit of the laboratory normal range. 
17.4. VITAL SIGNS 
The following vital sign parameters will be collected for this study as per the schedule of events (refer to protocol, 
Table 3): 
• 
Systolic blood pressure (SBP) (mmHg) 
• 
Diastolic blood pressure (DBP) (mmHg)  
• 
Pulse rate (beats per minute [bpm]) 
• 
Oxygen saturation (%) 
• 
Body temperature (⁰C) 


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• 
Weight (kg) 
The following summaries will be provided by treatment group based on the SAF analysis population for each vital 
sign parameter: 
• 
Observed and change from baseline by visit and timepoint 
• 
Number and percentages of subjects with at least one markedly abnormal post-baseline observed value/change 
from baseline (refer to Section 17.4.1) 
All vital sign data will be listed. Vitals signs meeting one of the markedly abnormal criteria in Section 17.4.1 will be 
identified. 
17.4.1. 
VITAL SIGNS MARKEDLY ABNORMAL CRITERIA 
Markedly abnormal vital sign observed values and/or change from baseline will be identified in accordance with the 
following predefined markedly abnormal criteria: 
Variable 
Unit 
Low 
High 
SBP 
mmHg 
≤ 90 mmHg AND  
change from baseline ≤ -20 mmHg 
≥ 180 mmHg AND 
change from baseline ≥ 20 mmHg 
DBP 
mmHg 
≤ 50 mmHg AND 
change from ≤ -15 mmHg 
≥ 105 mmHg AND 
change from baseline ≥ 15 mmHg 
Pulse rate 
bpm 
≤ 50 bpm AND 
change from baseline ≤ -15 bpm 
≥ 120 bpm AND 
change from baseline ≥ 15 bpm 
Oxygen 
saturation 
% 
< 94 % 
Not applicable 
Body 
temperature 
C 
Not applicable 
≥ 38.3 C AND 
change from baseline ≥ 1.1 C 
Weight 
kg 
Percent change from baseline  
≤ -7.0 % 
Percent change from baseline  
> 7.0 % 
 


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17.5. ECG EVALUATIONS 
The following electrocardiogram (ECG) parameters will be measured for this study as per the schedule of events 
(refer to protocol, Table 3): 
• 
PR interval (msec) 
• 
RR interval (msec) 
• 
QRS interval (msec) 
• 
QT (uncorrected) interval (msec) 
• 
Overall ECG evaluation (Investigator’s judgment) 
o 
Normal 
o 
Abnormal, not clinically significant (NCS) 
o 
Abnormal, clinically significant (CS) 
The following summaries will be provided by treatment group based on the SAF analysis population for each ECG 
parameter: 
• 
Observed and change from baseline by visit (for quantitative parameters) 
• 
Number and percentages of subjects with at least one markedly abnormal post-baseline observed value/change 
from baseline (for quantitative parameters; refer to Section 17.5.11) 
• 
Shift from baseline in overall ECG interpretation to the worst post-baseline assessment 
All ECG data will be listed. ECG results meeting one of the markedly abnormal criteria in Section 17.5.1 will be 
identified. 
17.5.1. 
ECG MARKEDLY ABNORMAL CRITERIA 
Markedly abnormal quantitative ECG parameters will be identified in accordance with the following predefined 
markedly abnormal criteria: 
• 
Observed values for QT (uncorrected) interval will be classified as: 


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o 
> 450 msec 
o 
> 480 msec 
o 
> 500 msec 
• 
Change from baseline for QT (uncorrected) interval will be classified as: 
o 
>30 msec increase from baseline 
o 
>60 msec increase from baseline 
It is to be noted that the previous categories are not mutually exclusive, but cumulative.  For example, if a subject 
worst post-baseline QT post-baseline observed value is 490 mmHg, then this subject will be reported once under QT 
> 450 msec and once under QT > 480 msec. 
18. DATA NOT SUMMARIZED OR PRESENTED 
Data that will not be summarized or listed are: 
• 
Results of serology tests performed (i.e., HIV, Hepatitis A, B, and C) at screening for the purpose of 
determining eligibility. 
 
 


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APPENDIX 1. PARTIAL DATE CONVENTIONS AND MISSING DATA 
IMPUTATION 
ALGORITHM FOR PRIOR ADVERSE EVENTS, TREATMENT-EMERGENT  ADVERSE 
EVENTS, AND POST-TREATMENT ADVERSE EVENTS 
START DATE 
END DATE 
ACTION 
Known 
Known/Partial/ 
Missing 
If (AE start date < study drug start date) or AE is a pre-
initial dose event, then PRAE. 
If study drug start date ≤ AE start date and AE start date <= 
last drug date_30 and AE is not a pre-initial dose event, then 
TEAE. 
If study drug start date <= AE start date and AE start date > 
last drug date + 30  then PTAE 
 
 
 
Partial, but known components 
show that AE started before 
study drug start date or AE is a 
pre-initial dose event 
Known/Partial/ 
Missing 
 
PRAE 
 
 
 
Partial and known components 
show that AE started on or after 
study drug start date  
 
Known/Partial/ 
Missing 
Impute missing AE start day as the first of the month or the 
first study drug dose date, whichever is later.  
If (AE start date < study drug start date) or AE is a pre-
initial dose event, then PRAE. 
If study drug start date ≤ AE start date and AE start date <= 
last drug date+30 and AE is not a pre-initial dose event, 
then TEAE. 
If study drug start date <= AE start date and AE start date > 
last drug date+30 then PTAE 
Note: If an AE started prior to first dose of study drug but worsened after treatment started, it will be reported as a 
separate adverse event with a new AE start date. A pre-initial dose event is defined as an AE that is entered in the 


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EDC as occurring prior to infusion (Month 0, Day 1). 
ALGORITHM FOR PRIOR / CONCOMITANT MEDICATIONS 
START DATE 
STOP DATE 
ACTION 
Known 
Known or 
ongoing 
If medication stop date < study drug start date, assign as prior; 
Otherwise, assign as concomitant.  
Partial 
If known components of medication stop date show that medication stopped 
before study drug start date, assign as prior; 
Otherwise, assign as concomitant. 
Missing, not 
ongoing 
Can never be assigned as prior, therefore assign as concomitant. 
 
 
 
Partial 
Known or 
ongoing 
If medication stop date < study drug start date, assign as prior; 
Otherwise, assign as concomitant. 
Partial 
If known components of medication stop date show that medication stopped 
before study drug start date, assign as prior; 
Otherwise, assign as concomitant. 
Missing, not 
ongoing 
Cannot be assigned as prior, therefore assign as concomitant. 
 
 
 
Missing 
Known or 
ongoing 
If medication stop date < study drug start date, assign as prior; 
Otherwise, assign as concomitant. 
Partial 
If known components of medication stop date show that medication stopped 
before study drug start date, assign as prior; 
Otherwise, assign as concomitant. 
Missing, not 
ongoing 
Assign as concomitant. 
 


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ALGORITHM FOR POTENCY ASSAY 
As specified in the inclusion criteria 3 in the protocol, a subject is eligible for enrollment as the study participant, 
only if the subject “has a CD8+ T cell defect that is correctable in vitro by co-culture with immature dendritic cells 
loaded with the synthetic oligopeptide from Hsp60sp”.  
 
Therefore, for the potency assay result of each patient on Visit 3/Day 1 (before the study treatment), should be 
imputed as the same as the corresponding potency assay result on Visit 1/Screening.  
 
For potency assay results that are missing on any visits other than Visit 3/Day 1, no imputation will be performed. 


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APPENDIX 2. LABORATORY ASSESSMENTS 
Chemistry (SI unit) 
 
• 
Blood urea nitrogen (BUN) (mmol/L) 
• 
Creatinine (µmol/L) 
• 
Calcium (mmol/L) 
• 
Chloride (mmol/L) 
• 
Potassium (mmol/L) 
• 
Sodium (mmol/L) 
• 
Bicarbonate (mmol/L) 
• 
Glucose (mmol/L) 
• 
Alkaline phosphatase (ALP) (U/L) 
 
• 
Alanine transaminase (ALT) (U/L) 
 
• 
Aspartate transaminase (AST) (U/L) 
 
• 
Albumin (g/L) 
• 
Total protein (g/L) 
• 
Total bilirubin (µmol/L) 
 
• 
HbA1c, L/L 
• 
Creatinine Clearance, mL/s 
 
 
 
 
 
 
 
 


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Hematology (SI unit) 
 
• 
Hemoglobin (g/L) 
 
• 
Hematocrit 
 
• 
Red blood cells (RBC) (x10E12/L) 
 
• 
White blood cell (WBC) count total (x10E9/L) 
 
• 
Platelets (x10E9/L) 
 
• 
Absolute neutrophils count (x10E9/L) 
 
• 
Absolute lymphocyte count (x10E9/L) 
 
• 
Absolute monocyte count (x10E9/L) 
 
• 
Absolute eosinophils count (x10E9/L) 
 
• 
Absolute basophils count (x10E9/L) 
 
 
 


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Urine Microalbumin Panel 
•
Urine Microalbumin (mg/dL)
•
Urine Creatinine (mg/dL)
•
Microalbumin/Creatinine Ratio (ratio)


Summary of Change, SAP v2.0 vs SAP v1.0 
•
Added supplementary analysis for Day 360
•
Deﬁned Change from Baseline at baseline, so that the baseline data can be included in MMRM
•
Deﬁned the imputaƟon rule for Potency Assay results on Day 1 (which was not planned to be
collected on Day 1 per protocol), based on the Potency Assay results from Screening
•
In addiƟon to Total Daily Insulin usage, also added Total Daily Insulin usage divided by weight
as a secondary endpoint
•
Added addiƟonal MMRM analysis for C-pepƟde AUC, HbA1c, Total Daily Insulin usage, and 4
auto-anƟbodies.