Effect of rhIL-7-hyFc on Increasing Lymphocyte Counts in Patients With Newly Diagnosed Non-severe Lymphopenic Gliomas Following Radiation and Temozolomide
Phase: PHASE1
Conditions: Glioma
Sponsor: Washington University School of Medicine

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Effect of rhIL-7-hyFc on increasing lymphocyte counts in patients with newly diagnosed 
non-severe lymphopenic gliomas following radiation and temozolomide 
 
 
Washington University School of Medicine 
Division of Oncology 
660 South Euclid Avenue, Campus Box #8056 
St. Louis, MO 63110 
 
Protocol #: 201810185 
Version date: 5/15/2023  
NCT #:  NCT03687957 
IND #: 141447 
 
Coordinating Center: 
Washington University School of Medicine 
 
PI: Milan G. Chheda, M.D. 
Associate Professor of Medicine and Neurology 
Office: 314-747-2712 
Email: mchheda@wustl.edu 
 
Sub-Investigators:  
 
Institution 
George Ansstas, M.D. 
 
Washington University  
Omar Butt, M.D., Ph.D. 
 
Washington University 
Jiayi Huang, M.D. 
 
 
Washington University 
Tanner Johanns, M.D., Ph.D.  
Washington University 
Richard Hotchkiss, M.D. 
 
Washington University 
John DiPersio, M.D., Ph.D. 
 
Washington University 
Jingqin Rosy Luo, Ph.D. 
 
Washington University 
Sonika Dahiya, M.D.  
 
Washington University 
 
 
 
 
CONFIDENTIAL 
The information contained in this document is regarded as confidential and, except to the extent 
necessary to obtain informed consent, may not be disclosed to another party unless law or 
regulations require such disclosure.  Persons to whom the information is disclosed must be 
informed that the information is confidential and may not be further disclosed by them 
 
 

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Effect of rhIL-7-hyFc on increasing lymphocyte counts in patients with newly diagnosed 
non-severe lymphopenic gliomas following radiation and temozolomide 
 
Protocol Revision History 
 
Initial Approval Version 
 
 
 
 
 
 
10/16/2018 
Amendment #1 Version 
 
 
 
 
 
 
11/07/2019 
Amendment #2 Version 
 
 
 
 
 
 
01/10/2019 
Amendment #3 Version 
 
 
 
 
 
 
04/16/2019 
Amendment #4 Version 
 
 
 
 
 
 
07/17/2019 
Amendment #5 Version 
 
 
 
 
 
 
12/13/2019 
Amendment #6 Version 
 
 
 
 
 
 
02/25/2020 
Amendment #7 Version 
 
 
 
 
 
 
03/04/2020 
Amendment #8 Version 
 
 
 
 
 
 
10/01/2020 
Amendment #9 Version 
 
 
 
 
 
 
02/08/2021 
Amendment #10 Version 
 
 
 
 
 
 
08/05/2021 
Amendment #11 Version 
 
 
 
 
 
 
10/12/2021 
Amendment #12 Version 
 
 
 
 
 
 
04/28/2022 
Amendment #13 Version 
 
 
 
 
 
 
07/18/2022 
Amendment #14 Version 
 
 
 
 
 
 
04/19/2023 
Amendment #15 Version 
 
 
 
 
 
 
05/15/2023 
 
 
 

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Effect of rhIL-7-hyFc on increasing lymphocyte counts in patients with newly diagnosed 
non-severe lymphopenic gliomas following radiation and temozolomide 
 
Principal Investigator Signature Page 
 
 
Principal Investigator 
(printed): 
      
Name of Institution:       
  
 PI Signature  
 
 
 
 
Date 
 
By my signature, I agree to personally supervise the conduct 
of this study and to ensure its conduct in compliance with the 
protocol, informed consent, IRB/HRPO procedures, the 
Declaration of Helsinki, ICH Good Clinical Practices 
guidelines, and the applicable parts of the United States 
Code of Federal Regulations or local regulations governing 
the conduct of clinical studies. 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

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Effect of rhIL-7-hyFc on increasing lymphocyte counts in patients with newly diagnosed 
non-severe lymphopenic gliomas following radiation and temozolomide 
 
SCHEMA 
 
 
Dose Level 
rhIL-7-hyFc Dose 
1 
60 mcg/kg 
2 
120 mcg/kg 
3 
240 mcg/kg 
4 
540 mcg/kg 
5 
720 mcg/kg 
6 
960 mcg/kg 
 
 
 
 

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N= 31 

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TABLE OF CONTENTS 
 
SCHEMA ................................................................................................................................... 4 
1 
BACKGROUND AND RATIONALE ..................................................................................... 8 
1.1 
Treatment-related lymphopenia and high grade gliomas .............................................. 8 
1.2 
Interleukin-7 and immune reconstitution ....................................................................... 8 
1.3 
IL-7 in GBM .................................................................................................................. 9 
1.4 
rhIL-7-hyFc .................................................................................................................10 
1.5 
Study rationale ............................................................................................................10 
1.6 
Protocol Amendment #3..............................................................................................10 
1.7 
Protocol Amendment #4..............................................................................................11 
1.8 
Protocol Amendment #11 ............................................................................................12 
2 
STUDY OBJECTIVES........................................................................................................12 
2.1 
Primary objectives.......................................................................................................12 
2.2 
Secondary objective ....................................................................................................12 
2.3 
Exploratory objectives .................................................................................................13 
3 
PATIENT ELIGIBILITY CRITERIA .....................................................................................13 
3.1 
Inclusion Criteria .........................................................................................................13 
3.2 
Exclusion Criteria ........................................................................................................13 
3.3 
Inclusion of Women and Minorities .............................................................................14 
4 
REGISTRATION PROCEDURES ......................................................................................14 
4.1 
Confirmation of Patient Eligibility .................................................................................14 
4.2 
Patient Registration in the Siteman Cancer Center OnCore Database ........................15 
4.3 
Assignment of UPN .....................................................................................................15 
4.4 
Randomization ............................................................................................................15 
5 
RESEARCH PLAN .............................................................................................................16 
5.1 
Premedications ...........................................................................................................16 
5.2 
Agent Administration ...................................................................................................16 
5.3 
Dose Escalation Schema (Phase I) .............................................................................17 
5.4 
Definition of MTD, DLT, and Dose Escalation Criteria .................................................17 
5.5 
Phase II and Expansion Cohort...................................................................................18 
5.6 
Toxicity, Response, and DLT Evaluations ...................................................................19 
5.7 
Women of Childbearing Potential ................................................................................19 
5.8 
Duration of Therapy ....................................................................................................19 
5.9 
Duration of Follow-up ..................................................................................................20 
6 
DOSE DELAYS / DOSE MODIFICATIONS ........................................................................20 
6.1 
Temozolomide Dose Modifications .............................................................................20 
6.2 
rhIL-7-hyFc Dose Modifications ...................................................................................20 
7 
REGULATORY AND REPORTING REQUIREMENTS ......................................................22 
7.1 
Sponsor-Investigator Reporting Requirements ............................................................22 
7.2 
Secondary Site Reporting Requirements ....................................................................24 
7.3 
Exceptions ..................................................................................................................25 
8 
PHARMACEUTICAL INFORMATION ................................................................................25 
8.1 
rhIL-7-hyFc .................................................................................................................25 
8.2 
rhIL-7hyFc Diluent (Will be used as Placebo in Phase II) ............................................26 
9 
CORRELATIVE STUDIES .................................................................................................27 

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9.1 
T lymphocyte subtypes and other PBMC subsets analysis .........................................27 
9.2 
T cell repertoire diversity .............................................................................................28 
9.3 
Cytokine analysis ........................................................................................................28 
9.4 
Immunogenicity ...........................................................................................................28 
9.5 
Tumor tissue analysis (optional) ..................................................................................28 
9.6 
Lymphocyte functional analysis (optional) ...................................................................28 
9.7 
Specimen Processing for PBMC .................................................................................29 
9.8 
Specimen Processing for ELISPOT (optional) .............................................................29 
9.9 
Anti-drug antibodies and neutralizing anti-drug antibodies (immunogenicity) ..............29 
10 
STUDY CALENDARS ....................................................................................................31 
10.1 Phase I and Randomized Phase II ..............................................................................31 
10.2 Phase II Expansion Cohort .........................................................................................32 
11 
DATA SUBMISSION SCHEDULE ..................................................................................34 
11.1 Adverse Event Collection in Case Report Forms ........................................................34 
12 
DATA AND SAFETY MONITORING ..............................................................................34 
13 
MEASUREMENT OF EFFECT .......................................................................................36 
13.1 rhIL-7-hyFc effect ........................................................................................................36 
13.2 Antitumor Effect – RANO ............................................................................................36 
13.3 Disease Parameters ...................................................................................................37 
13.4 Methods for Evaluation of Measurable Disease ..........................................................38 
14 
AUDITING ......................................................................................................................40 
15 
STATISTICAL CONSIDERATIONS ................................................................................41 
15.1 Study Design ..............................................................................................................41 
15.2 The phase I study .......................................................................................................41 
15.3 The randomized phase II study ...................................................................................43 
15.4 The expansion cohort .................................................................................................44 
15.5 Analysis set.................................................................................................................45 
15.6 Early stopping rules ....................................................................................................46 
15.7 Ad hoc proof-of-concept check....................................................................................46 
16 
MULTICENTER REGULATORY REQUIREMENTS .......................................................46 
17 
BIBLIOGRAPHY AND REFERENCES CITED ................................................................48 
APPENDIX A: NANO Scale ......................................................................................................52 
APPENDIX B: Safety Data ........................................................................................................54 
APPENDIX C: Definitions for Adverse Event Reporting ............................................................56 
APPENDIX D: Reporting Timelines ...........................................................................................58 
APPENDIX E: Washington University SAE Reporting Cover Sheet ..........................................61 
 
 

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1 
BACKGROUND AND RATIONALE  
 
1.1 
Treatment-related lymphopenia and high grade gliomas 
 
Patients with glioblastoma (GBM) routinely 
receive radiation (RT), temozolomide (TMZ), and 
glucocorticoids. However, these treatments cause 
severe (grade III-IV) and long lasting lymphopenia 
in 40% of patients. This treatment-related 
lymphopenia (TRL) (Fig. 1) is associated with 
shorter survival (Grossman, 2011) (Fig. 2). Any 
grade of lymphopenia was seen in up to ~75% of 
patients who were treated with RT and TMZ. The 
survival time is directly associated with the levels 
of CD4 and total lymphocyte counts. These data 
were derived from a prospective multi-center 
study with ~100 patients enrolled and it provides 
strong scientific rationale for this study. A similar 
relationship between TRL and shorter survival 
also exists in patients with newly diagnosed 
pancreatic, non-small cell lung cancer, and head 
and neck cancer (Grossman, 2015). Although 
data were obtained from retrospective analyses, these studies showed strikingly similar 
results. Thus these prior retrospective studies 
provide a strong scientific rationale for designing 
active interventions to enhance lymphocyte 
recovery or minimize TRL in an effort to improve 
outcomes 
for 
patients 
receiving 
chemo-
radiotherapy for GBM and other cancers. 
Recently, we reported that nearly all lymphocytes 
and lymphocyte subsets were significantly 
depressed after RT + TMZ, and remained at low 
levels for more than one year (Campian, 2017). 
Data from computer modeling studies suggest 
that a standard course of partial brain radiation is 
potentially lethal to over 95% of circulating 
lymphocytes (Yovino 2013). Recent data suggest 
that inadvertent radiation exposure to circulating 
lymphocytes passing through a treatment field 
could play a key role in TRL development in pancreatic cancer (Tang 2014; Wild 2013).  
 
It remains to be determined if the immune system can be restored after TRL-induced 
damage. Correction of TRL through an immunotherapeutic approach would potentially 
represent a major advance for the treatment of patients with GBM. However, the best 
way for restoring immune status has not been established. Here, we explore a novel 
approach to prevent or mitigate TRL in GBM in order to improve survival in these cancer 
patients.  
 
1.2 
Interleukin-7 and immune reconstitution 
 
Interleukin-7 (IL-7), a cytokine secreted by stromal cells in the bone marrow and thymus 
 
 
Fig. 1. CD4 Count Trend over Time. The 
median CD4 counts prior to chemoradiation 
were 664 cells/mm3. CD4 counts dropped 
significantly after 2 months. 73% had CD4 
counts <300 cells/mm340% had CD4 counts 
<200 cells/mm3. CD4 counts remained 
persistently low during the 12 months of 
 
Fig. 2. Overall survival by CD4 counts at 2 
months. Survival in patients with CD4 counts 
<200 cells/mm3 at 2 months was shorter than 
those with higher CD4 counts.  

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is required for human thymocyte development and for maintenance of mature peripheral 
lymphocyte  homeostasis.   In addition it is also important in both B cell and NK cell 
proliferation and survival. IL-7 plasma levels are inversely correlated with peripheral T 
CD4 cell counts. Although IL-7 is limited under normal conditions, it accumulates during 
lymphopenic conditions (Sportès 2008). IL-7 administration produces “new T cells”, and 
in preclinical studies IL-7 therapy significantly restored T cell immune function in mice 
(Mackall 1997) and primates (Beq  2006; Fry 2003; Storek 2003). Recent clinical trials 
using recombinant human interleukin (rhIL-7) have demonstrated its potential to expand 
and protect CD4 and CD8 T-cells (Mackall 2011). To date, rhIL-7 has been studied in 
multiple solid tumors with good tolerance and limited toxicity (Mackall 2011). The most 
common side effects reported for rhIL-7 therapy are low-grade fever, malaise, transient 
increases in liver enzyme levels, erythema and induration at the site of administration. 
Because IL-7 is a modest activator of effector T cells, which quickly internalize their IL-
7 receptors after binding (Sportès 2008), rhIL-7 does not induce cytokine storm 
syndrome. In addition, rhIL-7 preferentially expands naïve and memory T cells with a 
significant broadening of circulating T cell receptor (TCR) repertoire diversity (Sportès 
2008).  In vivo half-life of rhIL-7 has been noted to be very short as 6.5-9.8 hours limiting 
its clinical use (Sportès 2010).  In addition to its short in vivo half-life the availability of 
rhIL-7 has been limited to negligible for the past decade. The French pharmaceutical 
company Cytheris (CYT107) has been unable to produce sufficient rhIL-7 for clinical 
trials and in fact went out of business in 2013.  CYT107 was then taken by a US Biotech 
Revimmune Inc in 2014. However, the company has not been able to provide sufficient 
CYT107 due to financial difficulties. This has resulted in a relative and, at times, 
complete lack of rhIL-7 for clinical trials.  There is a lack of clinical grade rhIL-7, coupled 
with the absence of any pharmaceutical companies (up until now) interested in the 
clinical development of this cytokine. In addition, the very short half-life of the Cytheris 
rhIL-7 made it challenging to administer in out-patient settings. These provocative data 
underscore the need for alternative sources of more active and longer-acting IL-7 for 
human clinical trials.  In this study, we will be the first in the United States to test a novel 
long acting recombinant human IL-7 analogue, rhIL-7-hyFc (half-life is 48-92 hours) from 
NeoImmmuneTech Inc. (NIT) for the treatment of patients with high grade glioma (HGG).  
 
1.3 
IL-7 in GBM 
 
We recently reported that the levels of IL-7 were inappropriately low in patients with 
GBM treated with chemoradiation who developed lymphopenia (Campian 2017). 
Similar results were reported in another study (Ellsworth, 2014). These findings suggest 
that IL-7 did not increase appropriately to lymphopenia in these groups of patients. This 
finding raises the question of whether administration of exogenous IL-7 could improve 
CD4 counts and immune reconstitution in this patient population. 
 

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1.4 
rhIL-7-hyFc  
 
rhIL-7-hyFc 
is 
a 
long-acting 
cytokine 
consisting of recombinant human IL-7 fused 
to a hybrid Fc (hyFc) region of IgG (Fig. 3). A 
first-in-human clinical trial of rhIL-7-hyFc  
(Protocol No. GX-I7-HV-001) was performed 
as a single ascending dose escalation study 
in healthy normal volunteers by NIT and 
Genexine. rhIL-7-hyFc was well tolerated in 
all patients without any serious adverse 
events. 
Subcutaneous 
administration 
of 
increasing doses of rhIL-7-hyFc led to a 
dramatic increase in the proliferation markers of T cells and percentage of effector memory 
and central memory T cells within one week, returning to baseline in approximately three 
weeks. In addition, the increasing doses of rhIL-7-hyFc were associated with more rapid 
and robust increases in total lymphocyte counts and Tcm and Tem cell subsets. Currently, 
a Phase I study in solid tumors is ongoing in South Korean (Protocol No.: GX-17-CA-003). 
They have shown that rhIL-7-hyFc was safely administered at 60 mcg/kg, 120 mcg/kg, 
240 mcg/kg, 480 mcg/kg, 720 mcg/kg, 960 mcg/kg and 1200 mcg/kg dose levels without 
DLTs.  In Study GX-I7-CA-004 in GBM, no DLTs were observed at 840 mcg/kg.  Please 
refer to Appendix B for more detailed safety data. 
 
Risks associated with taking rhIL-7-hyFc based on clinical experience include skin toxicity, 
liver toxicity, headaches, and enlarged lymph nodes. 
 
1.5 
Study rationale 
 
Here we have developed a phase I/II clinical trial to evaluate the effect of rhIL-7-hyFc on 
lymphocyte counts in patients with HGG. 
 
A phase I study will test whether rhIL-7-hyFc can be safely administered to patients with 
HGG. Six doses of rhIL-7-hyFc will be tested using a mix of Accelerated Phase and 
standard 3+3 dose-escalation design. The phase II portion to test effect of rhIL-7-hyFc on 
lymphocyte counts will use placebo-controlled randomization in HGG patients whose 
treatment include the standard RT and TMZ.  
 
In order to better understand the impact and benefit of rhIL-7-hyFc  administration, we will 
collect peripheral blood and tumor tissue specimens (when possible) before and after rhIL-
7-hyFc treatment in all patients. Peripheral blood mononuclear cells (PBMC) will be 
collected and cryopreserved for analysis.  Lymphocyte counts and T cell subsets will be 
evaluated by Fluorescence Activated Cell Sorting (FACS). T cell repertoire diversity will 
be characterized using PCR based deep sequencing by ImmunoSEQ assays (Adaptive 
Biotechnologies, Seattle, WA). Immunogenicity such as the occurrence of anti-drug 
antibodies (ADA) and neutralizing ADA will be monitored in collaboration with NIT. 
 
1.6 
Protocol Amendment #3 
 
1.6.1 Synopsis  
 
In this amendment, the dose escalation schema was updated to remove dose level 
 
Fig. 3. IL-7hyFc protein structure 

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360 mcg/kg and update the maximum dose level to 960 mcg/kg. 
 
1.6.2 Rationale 
 
Currently, a Phase 1b trial of single-agent NT-I7 (rhIL-7-hyFc) is being conducted 
in patients with advanced solid tumors in Korea (Study No. GX-I7-CA-003) utilizing 
a 3+3 dose escalation approach to determine the Recommended Phase 2 Dose 
(RP2D). The dose level ranges for this study include 60 µg/kg, 120 µg/kg, 240 
µg/kg, 480 µg/kg, 720 µg/kg, 960 µg/kg and 1200 µg/kg given intramuscularly (IM) 
in multiple doses.  Thus far, the safety profile of NT-I7 in the study has been very 
similar to the safety profile observed in the healthy volunteers study. There have 
been no dose limiting toxicities (DLTs). The most common treatment-related 
adverse events observed to date were injection-site reactions, which were mild to 
moderate in severity and resolved within a few days, usually without treatment 
intervention. 
 
1.7 
Protocol Amendment #4 
 
1.7.1 Synopsis 
 
In this amendment, an additional 3 patients will be enrolled to the dose level 4, 540 
mcg/kg. 
 
1.7.2 Rationale 
 
Due to the AEs seen in the 3 patients enrolled at dose level 4 (540 mcg/kg), 3 
additional patients will be added to that dose level. One out of these patients 
experienced Grade 2 AEs and all 3 of these patients experienced Grade 1 AEs. 
The additional 3 patients will be monitored for toxicity.  
 
In addition, it has been observed in this study and others that patients have 
received a relatively high total dose due to high body weight. With this observation, 
NeoImmuneTech (NIT) has released a pharmacy manual with revised dose 
calculation instructions for obese patients. rhIL-7-hyFc dosing is determined by 
actual body weight for non-obese patients; for obese patients (BMI ≥ 30kg/m²), 
dosing will be determined by adjusted body weight (see below formula). Weight 
must be re-assessed prior to each rhIL-7-hyFc dosing; if the patient’s weight 
changes +/- 10% from the baseline weight measurement, rhIL-7-hyFc dose should 
be based on the new weight measurement.  
 
 
 
BMI= (Weight in Kg)/(Height in meters)² 
 
For obese patients (BMI ≥ 30kg/m²), proceed with the following steps:  
 
1. Determine Ideal Weight (1 kg = 2.2 lbs):  
 
Males: 50 kg + 2.3 kg x (inch over 5 feet)  
Females: 45.5 kg + 2.3 kg x (inch over 5 feet)  
(Patients less than 5 feet: subtract 2.3 kg/inch under 5 feet)  
 

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2. Determine Adjusted Body Weight:  
 
Ideal Weight + 0.4 x (actual weight – ideal weight) = Adjusted Body Weight  
 
This change will be reflected in the next pharmacy manual when revised. 
 
1.8 
Protocol Amendment #11 
 
1.8.1 Synopsis 
 
This amendment adds an expansion cohort of 31 patients to the experimental arm 
(TMZ + rhIL-7-hyFc).  
 
Amendment 11 added an additional 31 patients to the rhIL-7hyFc treatment arm 
as an expansion cohort to evaluate the PFS in patients with IDH wildtype and 
MGMT promoter unmethylated GBM. In the expansion cohort, labs will be drawn 
per SOC and no research specimens will be collected. As of Amendment 14, 
enrollment into the expansion cohort may begin prior to completion of the phase II 
enrollment. 
 
1.8.2 Rationale 
 
Nineteen patients were enrolled to the phase I portion of the study. Among those, 
12 patients were IDH wildtype with MGMT promoter unmethylated GBM. Their 
mPFS to date (as of 7/29/2021) is 11.53 months (range 5.13- 14.8+). The reported 
mPFS is 5.4 months for unfavorable molecular profile and 7.3 months for favorable 
molecular profile in MGMT promoter unmethylated GBM (reference: Gilbert MR, 
2014 NEJM, PMCID: PMC4201043). This preliminary efficacy data suggests a 
potential improvement in mPFS with the study treatment. 
 
The purpose of this amendment is to test whether there is a significant 
improvement in mPFS. 
 
 
2 
STUDY OBJECTIVES 
 
2.1 
Primary objectives 
 
1. Phase I: To evaluate the safety, tolerability, and to determine the maximal tolerated 
dose (MTD) of rhIL-7-hyFc in patients with glioma.  
2. Randomized Phase II: To test the effect of rhIL-7-hyFc on absolute lymphocyte count 
(ALC) compared to control. 
3. Phase II Expansion Cohort: To determine the mPFS of patients with gliomas that are 
IDH1 wildtype, as defined by negative immunohistochemistry using an R132H-specific 
antibody, and MGMT promotor unmethylated who receive rhIL-7-hyFc.   
 
2.2 
Secondary objective 
 
To evaluate anti-drug antibodies (phase I and randomized phase II only) 
  

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2.3 
Exploratory objectives  
 
These objectives apply to the phase I and randomized phase II portions of this study only. 
 
1. To evaluate the impact of ADA on ALC level 
2. To evaluate the effect of concurrent dexamethasone 
3. To evaluate the serial T cell lymphocyte subtypes over time 
4. To evaluate the serial cytokine levels   
5. To evaluate the impact of adjuvant temozolomide on rhIL-7-hyFc effects on ALC 
6. To monitor overall response rate (ORR), progression free survival (PFS) and overall 
survival 
7. To explore the impact on tumor infiltrating lymphocytes (TIL) after rhIL-7-hyFc 
treatment (When tumor tissue specimens are available). 
8. Functional analysis of T cells before and after rhIL-7-hyFc treatment 
 
 
3 
PATIENT ELIGIBILITY CRITERIA 
 
 
3.1 
Inclusion Criteria 
 
1. WHO grade III, grade IV, and high risk grade II gliomas that require RT and TMZ 
treatment. 
2. Phase 2 Expansion Cohort ONLY: Must be IDH1 wildtype, as defined by negative 
immunohistochemistry using an R132H-specific antibody, and MGMT promoter 
unmethylated glioblastoma multiforme (WHO grade IV). 
3. Post-operative treatment must have included radiation and TMZ. Prior Gliadel Wafers 
are allowed. Glucocorticoid therapy is allowed. TTF device is allowed. 
4. Adequate organ and marrow function defined as follows: 
a. Absolute neutrophil count ≥ 1,000/mcL 
b. Platelets ≥ 75,000/mcL 
c. Hemoglobin ≥ 8 g/dL 
d. Total bilirubin ≤ 3.0 x institutional upper limit of normal 
e. AST (SGOT)/ALT (SGPT) ≤ 3.0 × institutional upper limit of normal 
5. ALC ≥ 600/mcL (required for phase I and randomized phase II only) 
6. Karnofsky Performance Status (KPS) ≥ 60% (i.e. the patient must be able to care for 
himself/herself with occasional help from others).  
7. Able to provide written informed consent (or consent from a legally authorized 
representative). 
8. Women of childbearing potential must have a negative serum pregnancy test prior to 
study entry (within 14 days). Patients must be willing to be on adequate contraception 
during treatment as defined in Section 5.5. 
9. ≥ 18 years of age.  
 
3.2 
Exclusion Criteria 
 
1. Receiving any other investigational agents which may affect patient’s lymphocyte 
counts. 
2. Pregnant women are excluded from this study because rhIL-7-hyFc has not been 
evaluated regarding its potential for teratogenic or abortifacients effects.  There is 
a potential risk for adverse events in nursing infants secondary to treatment of the 

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mother with the study drug, breastfeeding should be discontinued if the mother is 
treated with rhIL-7-hyFc. 
3. Has an active viral infection requiring systemic treatment at screening.  
4. Has active autoimmune disease or syndrome (i.e. moderate or severe rheumatoid 
arthritis, moderate or severe psoriasis, multiple sclerosis, myasthenia gravis, 
Guillain-Barre syndrome, systemic lupus erythematosis, scleroderma, ulcerative 
colitis, Crohn’s disease, autoimmune hepatitis, granulomatosis with polyangiitis, 
etc.,) that requires systemic treatment at the time of screening. Replacement 
therapy (e.g., thyroxine, insulin, or physiologic corticosteroid replacement therapy 
for adrenal or pituitary insufficiency) is not considered a form of systemic treatment. 
Subjects are permitted to enroll if they have vitiligo, resolved childhood 
asthma/atopy, type I diabetes mellitus, residual hypothyroidism due to 
autoimmune condition only requiring hormone replacement, psoriasis not requiring 
systemic treatment, or conditions not expected to recur in the absence of an 
external trigger.  
5. Receipt of live attenuated vaccine within 30 days before the first dose of study 
treatment.  Examples of live vaccines include, but are not limited to, the following: 
measles, mumps, rubella, chicken pox, yellow fever, rabies, Bacillus Calmette-
Guérin (BCG), Zoster, and typhoid vaccine.  Seasonal influenza vaccines for 
injection are generally killed virus vaccines and are allowed; however, intranasal 
influenza vaccines (e.g. FluMist) are live attenuated vaccines and are not allowed. 
6. Has clinically significant cardiac enzymes ([Tnl or TnT] or CK) 
7. Patients with a clinically significant EKG on screening triggering a echocardiogram 
which is also clinically significant  
 
3.3 
Inclusion of Women and Minorities 
 
Both men and women and members of all races and ethnic groups are eligible for this trial. 
 
 
4 
REGISTRATION PROCEDURES 
 
Patients must not start any protocol intervention prior to registration through the Siteman 
Cancer Center. 
 
The following steps must be taken before registering patients to this study: 
 
1. Confirmation of patient eligibility by Washington University 
2. Registration of patient in the Siteman Cancer Center database 
3. Assignment of unique patient number (UPN) 
 
Once the patient has been entered in the Siteman Cancer Center OnCore database, the WUSM 
coordinator will forward verification of enrollment and the UPN via email. 
 
4.1 
Confirmation of Patient Eligibility 
 
Confirm patient eligibility by collecting the information listed below and scanning and 
emailing it to the research coordinator listed in the Siteman Cancer Center Clinical Trials 
Core Protocol Procedures for Secondary Sites packet at least one business day prior to 
registering patient: 

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1. Your name and contact information (telephone number, fax number, and email 
address) 
2. Your site PI’s name, the registering MD’s name, and your institution name 
3. Patient’s race, sex, and date of birth (DOB) 
4. Three letters (or two letters and a dash) for the patient’s initials 
5. Currently approved protocol version date 
6. Copy of signed consent form (patient name may be blacked out) 
7. Planned date of enrollment 
8. Completed eligibility checklist, signed and dated by a member of the study team 
9. Copy of appropriate source documentation confirming patient eligibility 
 
4.2 
Patient Registration in the Siteman Cancer Center OnCore Database 
 
Registrations may be submitted Monday through Friday between 8am and 5pm CT. 
Urgent late afternoon or early morning enrollments should be planned in advance and 
coordinated with the Washington University research coordinator.  Registration will be 
confirmed by the research coordinator or his/her delegate by email within one business 
day. Verification of eligibility and registration should be kept in the patient chart. 
 
All patients at all sites must be registered through the Siteman Cancer Center OnCore 
database at Washington University. 
 
4.3 
Assignment of UPN 
 
Each patient will be identified with a unique patient number (UPN) for this study. Patients 
will also be identified by first, middle, and last initials.  If the patient has no middle initial, a 
dash will be used on the case report forms (CRFs).  All data will be recorded with this 
identification number on the appropriate CRFs. 
 
4.4 
Randomization 
 
A total of 20 patients during phase II will be randomized 1:1 to receive either placebo or 
rhIL-7hyFc at the dose determined during phase I. Patients will be stratified by 
concomitant use of steroids (Yes/No). This study is double-blinded (participant, and 
physician/study coordinator are all blinded; pharmacist and study statistician are not 
blinded). Approximately equal number of patients will be assigned to each treatment 
group. The randomization will be performed in OnCore. Randomization will occur on 
eligible patients only and is auto generated after the eligibility is confirmed, the patient has 
an on-study date, and the strata steroid use (Yes/No) is confirmed; which will return either 
an ‘A’ or ‘B’ assignment. Only the pharmacist and statistician will know the treatment 
corresponding to each assignment. 
 
Patients enrolled to the phase II expansion cohort will not be randomized. 
 
 

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5 
RESEARCH PLAN 
 
The Phase I part will begin with an Accelerated Phase with 1 patient per cohort at the first 2 doses 
(60 mcg/kg and 120 mcg/kg), followed by a standard 3+3 design on the remaining four dose levels 
(see Table in Section 5.2), each with a cohort of 3 independent patients each time and with a 
maximum total of 6 patients. The dose escalation rules are summarized in Section 5.3.3. Patients 
enrolled in dose level 1 and dose level 2 (60 mcg/kg and 120 mcg/kg) will have the option to 
increase their dose to 240 mcg/kg if no DLTs are observed at the 240 mcg/kg dose level and they 
are still receiving treatment on study. 
 
The maximum tolerated dose (MTD) if no DLT at the highest dose level will be reviewed together 
with ALC and PK/PD data to determine a recommended phase 2 dose (RP2D) for use in the 
Phase II study. In the Phase II part of the study, 20 eligible patients will be randomized at a 1:1 
ratio to receive either rhIL-7-hyFc or placebo.  The MTD was determined to be 720 mcg/kg.  
 
CBC with differential (including ALC) and CD4 counts will be tested during RT and TMZ treatment 
as standard of care (SOC). Research blood will be collected prior to the initiation of the study 
treatment and at the selected time points for patients enrolled in phase I and randomized phase 
II.  No research blood will be collected for the expansion cohort.  
 
If ADA is observed in week 45, ADA levels need to be monitored every 2 months until it decreases 
to the basal level. The monitoring of ADA levels should not interrupt rhIL-7-hyFc administrations 
if patients do not experience DLTs.   
 
An additional 31 patients will be enrolled in a non-randomized fashion as an expansion cohort to 
receive 720 mcg/kg rhIL-7-hyfc.  These patients must have GBM that is IDH1 wildtype confirmed 
by negative immunohistochemistry using an IDH1 mutant-specific antibody, and MGMT promoter 
unmethylated.  These patients will have labs drawn as per SOC with no research collections.  
Enrollment into the expansion cohort may begin prior to completion of the phase II enrollment. 
 
5.1 
Premedications 
 
Benadryl 25-50 mg oral can be given 30-60 min prior to rhIL-7-hyFc injection as pre-
medication to prevent skin reactions.  This step is optional per patient and treating 
physician’s preference.   
 
5.2 
Agent Administration 
 
Per standard treatment, patients will receive concurrent RT + TMZ followed by adjuvant 
Concurrent 
RT+TMZ
Intramuscular rhIL-7-hyFc injection
Specimen collection
1
TMZ days 1-5 every 28 days
Ki67 test
2 3 4
8
1213
20
2425
16
37 weeks
Ki67 test
14

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TMZ on Days 1-5 of a 28-day cycle for a total of 6 cycles.  rhIL-7hyFc / placebo will be 
given by intramuscular injection starting at the end of RT + TMZ (within 14 days after last 
day of RT + TMZ).  The second injection will be administered 3-5 days after the last dose 
of cycle 3 TMZ treatment (~week 13 in treatment schema). The third injection will be given 
3-5 days after the last dose of cycle 6 TMZ treatment (~week 25 in treatment schema). 
Note that the second and third injections should be administered once between Day 3 
through 5 following the last dose of TMZ to achieve the strongest response (meaning that 
if TMZ is delayed, the rhIL-7-hyFc/placebo injections will be delayed as well).  The fourth 
injection (last injection in the study) will be given after completion of monthly TMZ (~Week 
37 as shown in Treatment Schema). A total of 4 doses of rhIL-7-hyFc injections are 
planned.  
 
5.3 
Dose Escalation Schema (Phase I) 
 
Dose Level 
rhIL-7-hyFc Dose 
(mcg/kg) 
1 
60 
2 
120  
3 
240  
4 
540  
5 
720  
6 
960 
 
Dose escalation will not occur until all patients in the cohort are 30 days out from their first 
dose of rhIL-7-hyFc and the Principal Investigator has been able to review all toxicities. 
 
5.4 
Definition of MTD, DLT, and Dose Escalation Criteria 
 
5.4.1 Definition of Maximum Tolerated Dose (MTD) 
 
The maximum tolerated dose (MTD) is defined as the dose of rhIL-7-hyFc that 
yields a dose limiting toxicity rate less than 33%. If an MTD is not reached, the 
highest administered dose (960 mcg/kg) will be the highest dose to test for safety. 
 
5.4.2 Dose Limiting Toxicities (DLTs) 
 
DLT will be defined as ≥ grade 3 AEs that occur within 30 days from the treatment 
start possibly related to the drug regimen, with severity graded according to the 
Common Terminology Criteria for Adverse Events (CTCAE) 5.0.  
 
DLT definition will have the following exceptions: 
• 
Grade 3 dermatologic AEs, including injection site reaction, that resolves 
to Grade ≤ 2 in ≤ 3 days with supportive care 
• 
transient Grade 3 influenza-like symptoms or pyrexia controlled with 
medical management 
• 
Grade 3 fatigue that resolves to Grade ≤ 2 in ≤ 3 days 

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• 
Grade 3 neurologic toxicity responding within ≤ 7 days to steroids, 
anticonvulsants, or electrolyte correction 
• 
Grade 3 laboratory abnormalities that are asymptomatic and considered by 
the investigator not to be clinically significant 
 
The DLT evaluation period is the first 30 days of therapy. The target DLT rate is 
<33%.  
 
5.4.3 Dose Escalation Criteria 
 
The starting dose level will be 60 mcg/kg, with one patient enrolled.  If no DLT is 
observed at the 60 mcg/kg dose level, a new patient will be enrolled to dose level 
2 (120 mcg/kg).  If no DLT is observed at that dose level, the study will proceed to 
the conventional 3+3 design and enroll 3 patients to dose level 3.  If any DLTs are 
observed in the single patient enrolled in either dose level 1 or dose level 2, the 
accelerated phase will convert to a conventional 3+3 design at the dose level 
where the DLT was observed and remain so for all the dose levels above.  If excess 
DLTs (≥ 2 out of a total of 6 patients at a given dose level) are observed at dose 
level 1, the study will halt. 
 
Dose escalations will proceed as follows:  
 
 
Number of Patients 
with DLT at a Given 
Dose Level 
Escalation Decision Rule 
0 out of 3 
Enter 3 patients at the next higher dose level. 
>2 
Dose escalation will be stopped.  Three (3) additional 
patients will be entered at the next lowest dose level if only 
3 patients were treated previously at that dose. 
1 out of 3 
Enter at least 3 more patients at this dose level. 
 
If 0 of these 3 patients experience DLT, proceed to the next 
dose level. 
 
If 1 or more of this group suffer DLT, then dose escalation 
is stopped.  Three (3) additional patients will be entered at 
the next lowest dose level if only 3 patients were treated 
previously at that dose. 
≥2 out of 6  
Dose escalation will stop and the prior dose level will be 
considered the MTD. This is generally the recommended 
phase 2 dose. At least 6 patients must be entered at the 
recommended phase 2 dose. 
 
5.5 
Phase II and Expansion Cohort 
 
Patients will be randomized to either rhIL-7-hyFc or placebo on a 1:1 basis in the phase II 
portion of the study.  Dosing will be administered as described in Section 5.2 at a dose of 
720 mcg/kg. 
 

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As part of an expansion cohort, an additional 31 patients with IDH1 wildtype confirmed by 
negative immunohistochemistry using an IDH1 mutant-specific antibody, and MGMT 
promoter unmethylated GBM will be enrolled, also at the 720 mcg/kg dose level.  These 
patients will not be randomized; they will all receive rhIL-7-hyFc and will also be treated 
as described in Section 5.2. Enrollment into the expansion cohort may begin prior to 
completion of the phase II enrollment. 
 
5.6 
Toxicity, Response, and DLT Evaluations 
 
All patients who receive any study treatment are evaluable for toxicity. Patients are 
evaluated from first receiving study treatment until a 30-day follow up after the conclusion 
of treatment or death. 
 
All patients are evaluable for disease response unless they discontinue treatment prior to 
having any disease assessment. 
 
A patient is evaluable for DLT assessment only during the first 30 days after the first dose 
of rhIL-7-hyFc treatment and only if enrolled in phase I of the study.   
 
Only patients enrolled to phase I or the randomized phase II portions of the study are 
evaluable for the secondary and exploratory objectives. 
 
5.7 
Women of Childbearing Potential 
 
Women of childbearing potential (defined as women with regular menses, women with 
amenorrhea, women with irregular cycles, women using a contraceptive method that 
precludes withdrawal bleeding, and women who have had a tubal ligation) are required to 
have a negative pregnancy test within 14 days prior to the first dose of the study agent. 
Patients must be willing to be on adequate contraception during treatment.  
 
Female and male patients (along with their female partners) are required to use two forms 
of acceptable contraception, including one barrier method, during participation in the study 
and for 90 days following the last dose of the study agent.  
 
If a patient is suspected of being pregnant, the study agent should be immediately 
discontinued.  In addition, a positive urine test must be confirmed by a serum pregnancy 
test.  If it is confirmed that the patient is not pregnant, the patient may resume dosing. 
 
If a female patient or female partner of a male patient becomes pregnant during therapy 
or within 90 days after the last dose of the study agent, the investigator must be notified 
in order to facilitate outcome follow-up. 
 
5.8 
Duration of Therapy 
 
If at any time the constraints of this protocol are considered to be detrimental to the 
patient’s health and/or the patient no longer wishes to continue protocol therapy, the 
protocol therapy will be discontinued and the reason(s) for discontinuation documented in 
the case report forms. 
 
In the absence of treatment delays due to adverse events, treatment may continue for up 
to 4 doses of rhIL-7-hyFc or until one of the following criteria applies: 

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• 
Documented and confirmed disease progression 
• 
Death 
• 
Adverse event(s) that, in the judgment of the investigator, may cause severe or 
permanent harm or that rule out continuation of study drug 
• 
General or specific changes in the patient’s condition render the patient unable to 
receive further treatment in the judgment of the investigator 
• 
Confirmed pregnancy 
• 
Serious non-compliance with the study protocol 
• 
Lost to follow-up 
• 
Patient withdraws consent 
• 
Investigator removes the patient from study 
• 
The Siteman Cancer Center decides to close the study 
 
Patients who prematurely discontinue treatment for any reason will be followed as 
indicated in the study calendar. 
 
5.9 
Duration of Follow-up 
 
Patients will only be off study at the time of death. All patients will be assessed for survival  
by phone call, clinic visit, or medical records (e.g. physician notes/laboratory results of 
clinic or hospital visit) at 2 years and 5 years following the end of study treatment. 
 
 
6 
DOSE DELAYS / DOSE MODIFICATIONS 
 
6.1 
Temozolomide Dose Modifications 
 
TMZ dose delays or modifications will be per SOC at the treating physician’s discretion. 
 
6.2 
rhIL-7-hyFc Dose Modifications 
 
 
Dose of rhIL-7-
hyFc 
Dose Level -1 
Dose Level -2 
Dose Level -3 
1st Reduction 
of  rhIL-7-hyFc 
2nd Reduction 
of  rhIL-7-hyFc 
3rd Reduction of 
rhIL-7-hyFc 
starting dose per 
protocol 
50% of the 
starting dose 
25% of the 
starting dose 
discontinue study 
drug 
 
Discontinuation Rules and Procedures for rhIL-7-hyFc-Related Adverse Events 
 
Toxicity 
Hold 
Treatment 
for Grade 
Guidance for 
Restarting 
Guidance for Permanent 
Discontinuation 
Supportive Care 
Guidelines 
Immune medicated 
Myocarditis 
2 - 4 
NA 
• 
Permanently 
discontinue 
• 
Initiate 
treatment as per 
institutional 
guidelines and 
consider 

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Toxicity 
Hold 
Treatment 
for Grade 
Guidance for 
Restarting 
Guidance for Permanent 
Discontinuation 
Supportive Care 
Guidelines 
antiarrhythmic 
drugs, 
temporary 
pacemaker, 
ECMO, or VAD 
as appropriate. 
AST/ALT or bilirubin 
increase/ immune-
related hepatitis 
2 
Monitor 
frequently 
Related:  reduce 
by 1 dose level 
Unrelated:  
same dose level 
• AST or ALT > 5 × ULN 
or total bilirubin > 3 × 
ULN 
• Toxicity fails to resolve 
within 12 weeks of last 
dose of rhIL-7-hyFc 
• Permanently 
discontinue for liver 
enzyme elevations 
meeting criteria for Hy's 
Law 
• Monitor LFT results 
frequently (consider 
weekly) 
• Treat with IV or oral 
corticosteroids 
3/4 
N/A 
• Permanently 
discontinue 
• Treat with IV 
corticosteroids for 
24 to 48 hours.  When 
symptoms resolve to 
≤ Grade 1, taper 
steroids over no less 
than 4 weeks 
Rash/dermatitis 
2 
Consider 
interrupting for 
Grade 2 for > 1 
week 
 
• Start topical 
emollients, mild-
strength corticosteroid 
cream  
3 
Toxicity resolves 
to Grade 0-1 
Related:  reduce 
by 1 dose level 
Unrelated:  
same dose level 
• Toxicity fails to resolve 
within 12 weeks of last 
dose or inability to 
reduce corticosteroids 
to ≤ 10 mg of 
prednisone or 
equivalent per day 
within 12 weeks 
• Start topical 
emollients, 
antihistamines, and 
high-strength 
corticosteroid cream 
• Consider to 
administer oral 
corticosteroids 
Rash/dermatitis 
4 
N/A 
• Permanently 
discontinue 
• Treat with IV steroids 
followed by high-dose 
oral steroids. When 
symptoms resolve to 
≤ Grade 1/2, taper 
steroids over no less 
than 4 weeks 
Other AEs (except 
lymphopenia) not 
otherwise listed 
2/3 
Toxicity resolves 
to Grade 0-1 
Related: reduce 
by 1 dose level 
Unrelated: same 
dose 
• Toxicity fails to resolve 
within 12 weeks  
• Any Grade 3 toxicity 
that recurs 
 
• Corticosteroids may 
be indicated 
 

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Toxicity 
Hold 
Treatment 
for Grade 
Guidance for 
Restarting 
Guidance for Permanent 
Discontinuation 
Supportive Care 
Guidelines 
4 
N/A 
• Permanently 
discontinue 
• Corticosteroids may 
be indicated 
 
 
7 
REGULATORY AND REPORTING REQUIREMENTS 
 
The entities providing oversight of safety and compliance with the protocol require reporting as 
outlined below. Please refer to Appendix C for definitions and Appendix D for a grid of reporting 
timelines. 
 
Adverse events will be tracked from start of treatment through 30 days after the end of study 
treatment.  All adverse events must be recorded on the toxicity tracking case report form (CRF) 
with the exception of: 
• 
Baseline adverse events, which shall be recorded on the medical history CRF 
• 
Clinical or laboratory adverse events that are grade 1 
• 
Lymphopenia  
 
Refer to the data submission schedule in Section 11 for instructions on the collection of AEs in 
the EDC. 
 
Reporting requirements for Washington University study team may be found in Section 7.1.  
Reporting requirements for secondary site study teams participating in Washington University-
coordinated research may be found in Section 7.2. 
 
 
7.1 
Sponsor-Investigator Reporting Requirements 
 
7.1.1 Reporting to the Human Research Protection Office (HRPO) at 
Washington University 
 
Reporting will be conducted in accordance with Washington University IRB 
Policies. 
 
Pre-approval of all protocol exceptions must be obtained prior to implementing the 
change. 
 
7.1.2 Reporting to the Quality Assurance and Safety Monitoring 
Committee (QASMC) at Washington University 
 
The Washington University Sponsor-Investigator is required to notify the QASMC 
of any unanticipated problems involving risks to participants or others occurring at 
WU or any BJH or SLCH institution that has been reported to and acknowledged 
by HRPO.  (Unanticipated problems reported to HRPO and withdrawn during the 
review process need not be reported to QASMC.) 
 
QASMC must be notified within 10 calendar days of receipt of IRB 
acknowledgment via email to qasmc@wustl.edu.  Submission to QASMC must 
include the myIRB form and any supporting documentation sent with the form. 

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For events that occur at secondary sites, the Washington University Sponsor- 
Investigator is required to notify the QASMC within 10 days of Washington 
University notification via email to qasmc@wustl.edu.  Submission to QASMC 
must include either the myIRB form and supporting documentation or (if not 
submitted to myIRB) the date of occurrence, description of the event, whether the 
event is described in the currently IRB approved materials, the event outcome, 
determination of relatedness, whether currently enrolled participants will be 
notified, and whether the informed consent document and/or any study procedures 
will be modified as a result of this event. 
 
7.1.3 Reporting to the FDA 
 
The conduct of the study will comply with all FDA safety reporting requirements.  
PLEASE NOTE THAT REPORTING REQUIREMENTS FOR THE FDA DIFFER 
FROM REPORTING REQUIREMENTS FOR HRPO/QASMC.  It is the 
responsibility of the Washington University Principal investigator to report to the 
FDA as follows: 
 
• 
Report any unexpected fatal or life-threatening suspected adverse reaction 
(refer to Appendix C for definitions) no later than 7 calendar days after initial 
receipt of the information.   
• 
Report a suspected adverse reaction that is both serious and unexpected 
(SUSAR, refer to Appendix C) no later than 15 calendar days after it is 
determined that the information qualifies for reporting.  Report an adverse 
event (refer to Appendix C) as a suspected adverse reaction only if there is 
evidence to suggest a causal relationship between the drug and the adverse 
event, such as: 
o A single occurrence of an event that is uncommon and known to be 
strongly associated with drug exposure 
o One or more occurrences of an event that is not commonly associated 
with drug exposure but is otherwise uncommon in the population 
exposed to the drug 
o An aggregate analysis of specific events observed in a clinical trial that 
indicates those events occur more frequently in the drug treatment 
group than in a concurrent or historical control group 
• 
Report any findings from epidemiological studies, pooled analysis of multiple 
studies, or clinical studies that suggest a significant risk in humans exposed to 
the drug no later than 15 calendar days after it is determined that the 
information qualifies for reporting 
• 
Report any findings from animal or in vitro testing that suggest significant risk 
in humans exposed to the drug no later than 15 calendar days after it is 
determined that the information qualifies for reporting 
• 
Report any clinically important increase in the rate of a serious suspected 
adverse reaction of that listed in the protocol or IB within 15 calendar days 
after it is determined that the information qualifies for reporting  
 
Submit each report as an IND safety report in a narrative format or on FDA Form 
3500A or in an electronic format that FDA can process, review, and archive.  Study 
teams must notify the Siteman Cancer Center Protocol Development team of each 

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potentially reportable event within 1 business day after initial receipt of the 
information, and must bring the signed 1571 and FDA Form 3500A to the Siteman 
Cancer Center Protocol Development team no later than 1 business day prior to 
the due date for reporting to the FDA. 
 
Each notification to FDA must bear prominent identification of its contents (“IND 
Safety Report”) and must be transmitted to the review division in the Center for 
Drug Evaluation and Research (CDER) or in the Center for Biologics Evaluation 
and Research (CBER) that has responsibility for review of the IND.  Relevant 
follow-up information to an IND safety report must be submitted as soon as the 
information is available and must be identified as such (“Follow-up IND Safety 
Report”). 
 
 
 
 
7.1.4 Reporting to NeoImmuneTech, Inc. 
 
All events being submitted to the FDA should be submitted concurrently to 
NeoImmuneTech by emailing them to Floryn Ajuzie, Clinical Trials Manager, at 
fajuzie@neoimmunetech.com and safety107@neoimmunetech.com. 
 
7.1.5 Reporting to Secondary Sites 
 
The Washington University Sponsor-Investigator (or designee) will notify the 
research team at each secondary site of all unanticipated problems involving risks 
to participants or others that have occurred at other sites within 10 working days 
of the occurrence of the event or notification of the Sponsor-Investigator (or 
designee) of the event.  This includes events that take place both at Washington 
University and at other secondary sites, if applicable. Refer to Section 16.0 
(Multicenter Management) for more information. 
 
7.2 
Secondary Site Reporting Requirements 
 
The research team at each secondary site is required to promptly notify the Washington 
University Sponsor-Investigator and designee of all serious adverse events (refer to 
Appendix C, Section D) within 1 working day of the occurrence of the event or notification 
of the secondary site’s PI of the event.  This notification may take place via email if there 
is not yet enough information for a formal written report (using either an FDA Form 3500a 
(MedWatch) and Washington University’s cover sheet (Appendix E)).  A formal written 
report must be sent to the Washington University Sponsor-Investigator and designee and 
research coordinator within 4 calendar days (for fatal or life-threatening suspected adverse 
reactions) or 11 calendar days (for serious unexpected adverse reactions) of the 
occurrence of the event or notification of the secondary site’s PI of the event. 
The research team at a secondary site is responsible for following its site’s guidelines for 
reporting applicable events to its site’s IRB according to its own institutional guidelines.  
The research team at Washington University is responsible for reporting all applicable 
events to the FDA and NeoImmuneTech as needed. 
Washington University pre-approval of all protocol exceptions must be obtained prior to 
implementing the change.  Local IRB approval must be obtained as per local guidelines.  

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Washington University IRB approval is not required for protocol exceptions occurring at 
secondary sites. 
 
7.3 
Exceptions  
 
Events that do not require expedited reporting as described in Section 7.1 include: 
• 
Planned hospitalizations 
• 
Hospitalizations < 24 hours 
• 
Respite care 
• 
Events related to disease progression 
 
Events that do not require expedited reporting must still be captured in the EDC. 
 
 
8 
PHARMACEUTICAL INFORMATION 
 
8.1 
rhIL-7-hyFc   
 
8.1.1 Description of Drug Substance  
 
rhIL-7-hyFc protein is produced by inserting the gene expressing rhrhIL-7-hyFc into 
the eukaryotic expression vector pAD15 at the Multiple Cloning Site. The Chinese 
Hamster Ovary (CHO) cell line DG44 is used to produce rhIL-7-hyFc.  
 
rhIL-7-hyFc has a molecular weight of 104 KDa and is composed of 400 amino acids 
with 155 amino acids for IL-7 and 30 for the IgD hinge, 8 for IgD the CH2 domain, 
and 207 for the IgG4 region. rhIL-7-hyFc contains 11 disulfide bonds, 1 O-
glycosylation, and 3 N-glycosylation sites. 
 
8.1.2 Chemical and Structural Formula 
 
Chemical Formula: C4012H6372N1104O1238S42 
 
Structural Formula: rhIL-7-hyFc is a fusion protein comprising human IL-7 fused to 
the human immunoglobulin D (IgD) hinge region. This in turn is fused to the N-
terminal region of CH2 from IgD and two key regions of the antibody immunoglobulin 
G4 (IgG4): C-terminal region of CH2, and the entire CH3 region. 
 
8.1.3 Pharmaceutical Properties 
 
rhIL-7-hyFc is supplied in a sterile, preservative-free liquid form in a single-use vial. 
One vial (1.1mL) contains 25 mg per 1 mL of the active ingredient of the finished drug 
product. The purity of the active ingredient must be 89.0% or higher based on size-
exclusion ultra-high-performance liquid chromatography (SE-UHPLC) testing and 
90.0% or higher based on reverse-phase high-performance liquid chromatography 
(RP-HPLC) testing. The rhIL-7-hyFc finished drug product should be a colorless, 
clear solution and should not contain any particulate matter that can be observed 
visually. 
 
8.1.4 Dosage Form  

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In addition to the active ingredient rhIL-7-hyFc, each vial contains sucrose, D-sorbitol, 
tri-sodium citrate dehydrates, citric acid monohydrate, and Polysorbate 80 as a 
stabilizer and buffer. These ingredients meet the specification criteria of the European 
pharmacopeia (Ph. EUR). 
 
rhIL-7-hyFc is supplied in a 1.1 mL vial package at a concentration of 25 mg 
protein/mL. The finished drug product solution contained in the vial is a liquid injection 
dosage form at pH 5.0±0.5 and a colorless, clear solution. There should not be any 
floating particulates under gross observation.  
 
rhIL-7-hyFc can be diluted with its diluent for intramuscular administration but should 
not be mixed with other drugs or solutions. Guidelines for dilution of rhIL-7-hyFc 
solution by hospital pharmacy are described in IB. 
 
8.1.5 Storage and Handling  
 
Vials that contain rhIL-7-hyFc must be kept refrigerated at 2∼8℃. The chemical and 
physical stability of the rhIL-7-hyFc finished drug product are being evaluated in an 
ongoing stability study at 2∼8℃. It is recommended that vials are protected from direct 
light until the time of use. 
 
8.1.6 Route of Administration 
 
Intramuscular injection. Do not shake vials before injection. rhIL-7-hyFc should be 
administered slowly in the intramuscular space. A vial is restricted to one patient and 
to one day of treatment. 
 
8.2 
rhIL-7hyFc Diluent (Will be used as Placebo in Phase II) 
 
8.2.1 Pharmaceutical Properties 
 
rhIL-7hyFc Diluent is supplied in a sterile, preservative-free liquid form, filled in the 
amount of 1.0 mL in a single-use, 2 mL glass vial. One vial contains the same 
ingredients with no active ingredient as the rhIL-7hyFc Drug Product. The rhIL-7-hyFc 
Diluent finished drug product should be a colorless, clear solution and should not 
contain any particulate matter that can be observed visually. 
 
8.2.2 Dosage Form  
 
Each vial contains sucrose, D-sorbitol, tri-sodium citrate dehydrates, citric acid 
monohydrate, and Polysorbate 80 as a stabilizer and buffer. These ingredients meet 
the specification criteria of the European pharmacopeia (Ph. EUR). 
 
Composition of rhIL-7-hyFc Diluent 
Component 
Reference to 
standards 
Function 
Quantity 
per mL 
Tri-Sodium citrate 
dihydrate 
Ph.Eur. 
pH adjusting 
agent 
3.68 mg 

Version 05/15/2023 
 
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Citric acid 
monohydrate 
Ph.Eur. 
pH adjusting 
agent 
1.58 mg 
Sucrose 
Ph.Eur. 
Osmolality 
modifying agent 
50.00 mg 
D-Sorbitol 
Ph.Eur. 
Osmolality 
modifying agent 
15.00 mg 
Polysorbate 80 
Ph.Eur. 
Surfactant 
0.5 mg 
Water for 
injection 
Ph.Eur. 
Solvent 
q.s. 1 mL 
 
rhIL-7-hyFc Diluent is supplied in a 2 mL vial package filled with 1 mL. The finished 
drug product solution contained in the vial is a liquid injection dosage form at pH 
5.0±0.5, osmolality is 300±40 mOsm/kg, and a colorless, clear solution. There should 
not be any floating particulates under gross observation.  
 
 
8.2.3 Storage and Handling  
 
Vials must be kept refrigerated at 2∼8℃. The chemical and physical stability of the 
rhIL-7-hyFc Diluent finished drug product are being evaluated in an ongoing stability 
study at 2∼8℃. It is recommended that vials be protected from direct light until the 
time of use. 
 
8.2.4 Route of Administration 
 
Intramuscular injection. Do not shake vials before injection. rhIL-7-hyFc Diluent 
should be administered slowly in the intramuscular space. A vial is restricted to 
one patient and to one day of treatment. 
 
 
9 
CORRELATIVE STUDIES 
 
NOTE: only patients enrolled to phase I and the randomized portion of phase II will have blood 
drawn for the correlative studies.  Patients enrolled to the expansion cohort will not have research 
blood drawn. 
 
9.1 
T lymphocyte subtypes and other PBMC subsets analysis 
 
The effect of rhIL-7-hyFc on immune cell phenotype and function in HGG patients is 
unknown. T lymphocyte absolute number and phenotype (i.e., CD4+ and CD8+ T cell 
naïve, effector memory, and central memory subsets, CD4+ Treg and activated CD4+ and 
CD8+ T cells) will be assessed using a 21-color, whole blood PBMC immunophenotyping 
assay as reported by the DiPersio laboratory (Staser 2017). The frequency and 
percentage of these and other PBMC subsets (i.e., T cells, NK cells, NKT cells, B cells, 
monocytes, dendritic cell subsets, and MDSC subsets) will also be investigated. The 
absolute change in each parameter, as well as variance in each parameter over time for 
each patient (mean, median, and SE/SD), will be evaluated. In addition, co-stimulatory 
and co-inhibitory receptors (PD-1, PD-L1, ICOS, TIM3, and LAG3) and T cell trafficking 
molecules (CXCR3, CCR5, LFA-1 and VLA-4) will be analyzed on cryopreserved PBMC.  
Cell proliferation Ki67 will be tested at the selected time shown in Treatment Schema.  

Version 05/15/2023 
 
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Additional markers delineating T cell exhaustion and activation, or other parameters may 
be analyzed on cryopreserved PBMC.   
 
 
9.2 
T cell repertoire diversity 
 
Because IL-7 receptor expression is high on resting, naive and memory populations, we 
expect that the rhIL-7-hyFc induced increase in circulating T cells will preferentially expand 
naive cells, and therefore lead to an overall increase in TCR diversity.  To evaluate the 
diversity of the T cell receptor repertoire, we will collect PBMC to characterize using TCR 
deep sequencing and analyze with Immunoseq Analyzer software (Adaptive 
Biotechnologies, Seattle, WA). Blood samples for T-cell repertoire analysis will be 
obtained at study week 1 (prior to the 1st injection of rhIL-7-hyFc; at study week 4; and 
then at study week 13 (prior to rIL-7-hyFc injection). 
 
 
9.3 
Cytokine analysis 
 
We will measure serum cytokine levels such as IL-7, IL-6, IL-15, INF-γ, TNF-α and TGF-
β using ELISA or Luminex, with modification to previously described methods (Levy 2012; 
Francois 2018) during the study. Plasma samples will be collected and stored for all 
subjects. 
 
9.4 
Immunogenicity  
 
Anti-drug antibodies have been reported in patients after one dose of rhIL-7-hyFc. The 
immunogenicity frequency and the occurrence of anti-drug antibodies (ADA) and 
neutralizing anti-drug antibodies (NADA) could be higher with exposure to multiple doses 
of rhIL-7-hyFc. However, presence of ADA and NADA was not associated with an increase 
in the frequency or severity of AEs. We will collaborate with NIT to monitor immunogenicity 
in patients treated with rhIL-7-hyFc by monitoring the formation of ADA and NADA to rhIL-
7-hyFc in serum. (Detail see below) 
 
9.5 
Tumor tissue analysis (optional) 
 
Whenever 
it 
is 
available, 
pre- 
and 
post-treatment 
formalin-fixed, 
paraffin-
embedded (FFPE) specimens will be examined to estimate CD8 and CD4+ TILs using 
immunohistochemistry analysis. As a complimentary method, using a new tissue-profiling 
method by Multiplex Immunofluorescence (GE Cell DiveTM system) (Sood 2016) that Drs. 
John DiPersio and James Hsieh recently installed in the department, we will assess the 
spatial distribution of immune cells at the tumor site. When possible, fresh tissue can be 
collected and saved to TPC for future analysis. 
 
9.6 
Lymphocyte functional analysis (optional) 
 
The ELISPOT assay will be used to determine the number of T-cells secreting IFN-γ in 
specific response to a viral antigen. Commercially available kits for virus-specific ELISPOT 
assays will be used and the number of spots will be read on an ELISPOT reader. 
 

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9.7 
Specimen Processing for PBMC 
 
A total of 32-40 mL of whole blood will be collected in 4-5 EDTA tubes for isolation of 
PBMCs at each of the following time points (NOTE-: +/- 7 days are allowed):   
• 
Week 1 (prior to the 1st dose of rhIL-7-hyFc) – 40 mL (32 mL for FACS, 8 mL 
for TCR sequencing). 
• 
Week 2 (one week after rhIL-7-hyFc) – 32 mL 
• 
Week 3 (two weeks after rhIL-7-hyFc) – 32 mL 
• 
Week 4 (three weeks after rhIL-7-hyFc) – 40 mL (32 mL for FACS, 8 mL for 
TCR sequencing). 
• 
Week 13 (prior to the 2nd dose of rhIL-7-hyFc) – 40 mL (32 mL for FACS, 8 
mL for TCR sequencing). 
• 
Week 14 (one week after rhIL-7-hyFc ) – 32 mL 
• 
At the time of tumor progression – 32 mL for FACS 
 
In addition, 2 mL of plasma will be saved during the isolation of PBMC. Saved plasma will 
be used for cytokine analysis. 
 
9.7.1 Handling of Specimens 
 
A total of 32-40 mL of whole blood will be collected in EDTA tubes and delivered 
to the DiPersio lab at room temperature within 2-hour of collection. PBMCs will be 
collected by density gradient centrifugation (Ficoll) per standard protocol. PBMC 
will be saved in -80°C for immunologic studies.  Plasma will be saved in -80°C for 
cytokine analysis. 
 
9.8 
Specimen Processing for ELISPOT (optional) 
 
A total of 5 mL of whole blood will be collected in one green top heparin tube at each of 
the following time points (NOTE-: +/- 7 days are allowed):   
• 
Week 1 (prior to the 1st dose of rhIL-7-hyFc) (as baseline) – 5 mL  
• 
Week 4 (three weeks after rhIL-7-hyFc) – 5 mL 
• 
Week 13 (prior to the 2nd dose of rhIL-7-hyFc) – 5 mL  
• 
Week 16 (three weeks after rhIL-7-hyFc ) – 5 mL 
• 
At the time of tumor progression – 5 mL  
 
9.8.1 Handling of Specimens 
 
A total of 5 mL of whole blood will be collected in heparin tube and delivered to the 
Hotchkiss lab or Dr. Dinesh Thotala’s lab at room temperature within 2-hour of 
collection. ELISPOT will be performed per lab standard procedure. 
 
9.9 
Anti-drug antibodies and neutralizing anti-drug antibodies 
(immunogenicity) 
 
The formation of anti-drug antibodies (ADA) and neutralizing anti-drug antibodies (NADA) 
to rhIL-7-hyFc will be evaluated: BioAgilytix will perform both Elisa Binding (non-
neutralizing) and neutralizing antibody assays according to their Standard Operating 
Procedure.  Serum from the above time points will be collected for these assays. 
 

Version 05/15/2023 
 
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NeoImmuneTech transferred the validated assay method to BioAgilytix who will perform 
the assays and the determination of the main ADA/NADA parameters. 
 
Collection of Specimen(s): Blood will be collected in serum separator tubes (SST) as 
follows: 
• 
Week 1 (prior to injection of 1st dose of rhIL-7-hyFc) – 8 mL 
• 
Week 13 (prior to the 2nd dose of rhIL-7-hyFc) – 8 mL 
• 
Week 45 (8 weeks after the last dose of rhIL-7-hyFc) – 8 mL 
• 
If ADA or NADA is observed in week 45, additional blood collections will be 
required every 2 months in order to monitor ADA/NADA levels until it decreases 
to the basal level.  
 
To date substantial amount of ADA data was obtained from the ongoing trials in Korea 
and the USA in solid tumors and GBM. The current data obtained indicates that the 
presence of ADA does not seem to affect drug exposure or pharmacovigilance data and 
safety profile of the drug. 
 
Handling of Specimens(s): For all samples, draw blood into SST tubes. Mix the content of 
the tube by slowly inverting several times (5 to 6 times) immediately after the blood draw, 
then sit at ambient temperature for 30-60 minutes until a clot has formed. Within 15 
minutes of blood collection, centrifuge the sample at room temperature at 2000 rpm for 5 
minutes.  The supernatant (serum) will be collected carefully for immediate storage and 
aliquot each serum sample into 4 cryovials, each of them containing at least 600 μL. 
Prepare appropriate labels and write the patient ID and time of preparation using a 
cryopen. Affix the labels to the cryovials. Freeze the samples in an upright position and 
store at -20ºC until shipment. 
 
Shipping of Specimen(s): Specimen should be packed on dry ice and shipped frozen in 
batches to the BioAgilytix (Kathryn Lindley, Director, Bioanalytical Operations, BioAgilytix, 
2300 Englert Dr., Durham, NC, 27713 USA. Email: kathryn.lindley@bioagilytix.com; 
Telephone: 919-381-6097, Monday-Thursday 9:00AM-4:30PM EST; shipments should be 
made Monday through Wednesday, or at least 2 days before any official holiday.)  
 
Site Performing Correlative Study: The ADA assay will be conducted by BioAgilytix, 
Durham, NC   
 
 

Version 05/15/2023 
 
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10 
STUDY CALENDARS 
 
10.1 
Phase I and Randomized Phase II 
 
Screening / baseline evaluations are to be conducted within 2 weeks prior to the start of protocol therapy.  Screening labs are used for 
screening purposes only and Baseline lab values will be considered those obtained on Week 1 Day 1, prior to first study dose injection.  
Unless otherwise noted study visits will be completed within a ± 1 week window. 
 
 
Screening 
W14 
W2 
W3 
W4 
W8 
W12 
W13 
W14 
W16 
W20 
W24 
W25 
W26 
W37 
W41 
W45 
FU9 
Confirm eligibility 
X13 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Informed consent 
X5 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
KPS 
X 
X 
 
 
 
 
X 
X 
 
 
 
 
X 
 
X 
 
 
 
Physical exam 
X 
X 
 
 
 
 
X 
X 
 
 
 
 
X 
 
X 
 
 
 
CBC w/diff incl. ALC8 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
 
X 
 
CD4 
X 
X 
 
 
X 
X 
X 
 
 
X 
X 
X 
 
 
 
 
 
 
LFTs 
X 
X11 
 
 
X 
 
X11 
 
 
X 
 
X11 
 
 
X11 
 
X 
 
EKG17 
X 
X 
 
 
 
 
 
X 
 
 
 
 
X 
 
X 
 
 
 
Cardiac Enzymes (Tnl 
or TNT and CK)17 
X 
X20 
 
 
 
 
 
X20 
 
 
 
 
X20 
 
X20 
 
 
 
Pregnancy test1 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Brain MRI18 
 
SOC MRIs (Q2-3Mo) 
Randomization2 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
NANO  
X 
 
 
 
 
 
X 
 
 
 
 
 
 
 
X 
 
 
 
rhIL-7-hyFc / placebo4 
 
X 
 
 
 
 
 
X 
 
 
 
 
X 
 
X 
 
 
 
SOC adjuvant TMZ3, 
14,15 
 
 
 
 
X 
X 
X 
 
 
X 
X 
X 
 
 
 
 
 
 
TMZ drug diary 
 
 
 
 
X 
X 
X 
 
 
X 
X 
X 
 
 
 
 
 
 
Ki67 
 
 
X 
 
 
 
 
 
X 
 
 
 
 
 
 
 
 
 
Research blood for 
PBMCs and plasma16 
 
X6 
X 
X 
X 
 
 
X6 
X 
 
 
 
 
 
 
 
 
 
Research blood for 
ADA and NADA 
 
X6 
 
 
 
 
 
X6 
 
 
 
 
 
 
 
 
X6 
X10 
OPTIONAL research 
blood for ELISPOT16 
 
X 
 
 
X 
 
 
X 
 
X 
 
 
 
 
 
 
 
 
OPTIONAL research 
tissue7 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
X12 
 
 
 
Survival 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
X 
AE assessment19 
 
X --------------------------------------------------------------------------------------------------------------------------------------------------- X 
 
 
1. Women of childbearing potential only 
2. Phase II only 
3. Given on Days 1-5 of a 28-day cycle for a total of 6 cycles. 

Version 05/15/2023 
 
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4. Within 14 days after the end of SOC RT/TMZ  
5. Does not have to occur within 2 weeks prior to the start of protocol therapy; participants may be consented as early as the start of standard of care treatment. 
6. Before injection 
7. If available 
8. Weekly CBC will be performed as per SOC and results will be collected for this study.  Weekly CBC will not be performed specifically for this study.  It may be drawn at a local lab. 
9. Assess survival at 2 years and 5 years following the end of treatment by chart review, phone call, or clinic visit. 
10. If ADA or NADA is observed, additional blood collections will be required Q2Mo to monitor ADA/NADA levels until it decreases to the basal level 
11. LFTs must be assessed no more than 7 days prior to each dose of rhIL-7-hyFc/placebo. 
12. At end of treatment. 
13. Eligibility to receive first study dose injection during Week 1 Day 1 will be determined by lab values obtained at Screening, and not reliant on lab values obtained on Day 1 
14. In the event of a delay, SOC labs (CBC, LFTs, and/or CD4) coinciding with visits of SOC TMZ dispensing will only be re-drawn if deemed medically necessary per treating physician 
discretion. 
15. Any delays in TMZ cycle dosing are permitted per SOC guidelines and per treating physician discretion and may not be indicative of exact weeks noted in study calendar. 
16. At the time of tumor progression 
17.  If a patient has an EKG that is deemed clinically significant (e.g., sinus tachycardia would not meet this definition, and non-specific changes may or may not meet this definition), 
then an echocardiogram should be performed. If an echocardiogram is performed and is deemed not clinically significant, the patient may be enrolled/continue in the study, with cardiac 
enzymes drawn as noted in footnote 20.  
18. MRI window is ±30 days 
19. AE’s will be followed through 30 days after the end of study treatment 
20. Only for patients with a clinically significant EKG  
 
10.2 
Phase II Expansion Cohort 
 
Screening / baseline evaluations are to be conducted within 2 weeks prior to the start of protocol therapy.  Screening labs are used for 
screening purposes only and Baseline lab values will be considered those obtained on Week 1 Day 1, prior to first study dose injection. 
Unless otherwise noted study visits will be completed within a ± 1 week window. 
 
 
Screening 
W14 
W2 
W3 
W4 
W8 
W12 
W13 
W14 
W16 
W20 
W24 
W25 
W26 
W37 
W41 
W45 
FU6 
Confirm eligibility 
X8 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Informed consent 
X4 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
KPS 
X 
X 
 
 
 
 
X 
X 
 
 
 
 
X 
 
X 
 
 
 
Physical exam 
X 
X 
 
 
 
 
X 
X 
 
 
 
 
X 
 
X 
 
 
 
CBC w/diff incl. ALC5 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
 
X 
 
CD4 
X 
X 
 
 
X 
X 
X 
 
 
X 
X 
X 
 
 
 
 
 
 
LFTs 
X 
X7 
 
 
X 
 
X7 
 
 
X 
 
X7 
 
 
X7 
 
X 
 
EKG11 
X 
X 
 
 
 
 
 
X 
 
 
 
 
X 
 
X 
 
 
 
Cardiac Enzymes (Tnl 
or TNT and CK)11 
X 
X14 
 
 
 
 
 
X14 
 
 
 
 
X14 
 
X14 
 
 
 
Pregnancy test1 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Brain MRI12 
 
SOC MRIs (Q2-3Mo) 
NANO  
X 
 
 
 
 
 
X 
 
 
 
 
 
 
 
X 
 
 
 
rhIL-7-hyFc 3 
 
X 
 
 
 
 
 
X 
 
 
 
 
X 
 
X 
 
 
 
SOC adjuvant TMZ2, 
9,10 
 
 
 
 
X 
X 
X 
 
 
X 
X 
X 
 
 
 
 
 
 
TMZ drug diary 
 
 
 
 
X 
X 
X 
 
 
X 
X 
X 
 
 
 
 
 
 

Version 05/15/2023 
 
page 33 of 61 
Survival 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
X 
AE assessment13 
 
X --------------------------------------------------------------------------------------------------------------------------------------------------- X 
 
 
1. Women of childbearing potential only 
2. Given on Days 1-5 of a 28-day cycle for a total of 6 cycles. 
3. Within 14 days after the end of SOC RT/TMZ  
4. Does not have to occur within 2 weeks prior to the start of protocol therapy; participants may be consented as early as the start of standard of care treatment. 
5. Weekly CBC will be performed as per SOC and results will be collected for this study.  Weekly CBC will not be performed specifically for this study.  They may be drawn at a local lab. 
6. Assess survival at 2 years and 5 years following the end of treatment by chart review, phone call, or clinic visit. 
7. LFTs must be assessed no more than 7 days prior to each dose of rhIL-7-hyFc/placebo. 
8. Eligibility to receive first study dose injection during Week 1 Day 1 will be determined by lab values obtained at Screening, and not reliant on lab values obtained on Day 1 
9. In the event of a delay, SOC labs (CBC, LFTs, and/or CD4) coinciding with visits of SOC TMZ dispensing will only be re-drawn if deemed medically necessary per treating physician 
discretion. 
10. Any delays in TMZ cycle dosing are permitted per SOC guidelines and per treating physician discretion and may not be indicative of exact weeks noted in study calendar. 
11. If a patient has an EKG that is deemed clinically significant (e.g., sinus tachycardia would not meet this definition, and non-specific changes may or may not meet this definition), 
then an echocardiogram should be performed. If an echocardiogram is performed and is deemed not clinically significant, the patient may be enrolled/continue on study with cardiac 
enzymes drawn as noted in footnote 14.  
12. MRI window is ±30 days 
13. AE’s will be followed through 30 days after the end of study treatment 
14.  Only for patients with a clinically significant EKG  
 
 
 

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11 
DATA SUBMISSION SCHEDULE 
 
Case report forms with appropriate source documentation will be completed according to the 
schedule listed in this section. 
 
Case Report Form 
Submission Schedule 
Original Consent Form 
Prior to registration 
On-Study Form 
Treatment Assignment Form 
Medical History Form 
Prior to starting treatment 
rhIL-7-hyFc Treatment Form 
Every treatment cycle (at W1, W12(Cycle 3), 
Week 24 (Cycle 6) and Week 37 
CBC Form 
As per SOC (see study calendar) 
NANO Form 
Response Form 
To coincide with SOC MRIs (Q2-3 months) 
DLT Form 
After Cycle 1 for phase I patients only 
SOC Treatment Summary Form At the end of SOC chemoradiation 
Research Blood Form 
Research Tissue Form 
Per Section 9.7 (phase I and randomized phase II 
patients only) 
Toxicity Form 
Steroid Form 
Continuous 
Treatment Summary Form 
Completion of treatment 
Follow Up Form 
Eat Year 2 and Year 5 Follow UP 
Progression Form 
Time of disease progression (not completed for 
expansion cohort) 
Death Form 
Time of death 
MedWatch Form 
See Section 7.0 for reporting requirements 
 
Any queries generated by Washington University must be responded to within 28 days of receipt 
by the participating site.  The Washington University research team will conduct a regular review 
of data status at all secondary sites, with appropriate corrective action to be requested as needed. 
 
11.1 
Adverse Event Collection in Case Report Forms 
 
All adverse events that occur beginning with start of treatment (minus exceptions defined 
in Section 7) must be captured in the Toxicity Form. Baseline AEs should be captured on 
the Medical History Form.  
 
Participant death due to disease progression should be reported on the Toxicity Form as 
grade 5 disease progression. If death is due to an AE (e.g. cardiac disorders; cardiac 
arrest), report as a grade 5 event under that AE. Participant death must also be recorded 
on the Death Form. 
 
 
12 
DATA AND SAFETY MONITORING 
 
In compliance with the Washington University Institutional Data and Safety Monitoring Plan, an 
independent Data and Safety Monitoring Board (DSMB) will be specifically convened for this trial 
to review toxicity data.  A DSMB will consist of no fewer than 3 members including 2 clinical 

Version 05/15/2023 
 
page 35 of 61 
investigators and a biostatistician.  DSMB members must be employed by Washington University, 
Barnes-Jewish Hospital, or St. Louis Children’s Hospital.  Like investigators, DSMB members are 
subject to the Washington University School of Medicine policies regarding standards of conduct. 
Individuals invited to serve on the DSMB will disclose any potential conflicts of interest to the trial 
principal investigator and/or appropriate university officials, in accordance with institution policies. 
Potential conflicts that develop during a trial or a member’s tenure on a DSMB must also be 
disclosed.  
 
Until such a time as the first secondary site enrolls its first patient, a semi-annual DSM report to 
be prepared by the study team will be submitted to the Quality Assurance and Safety Monitoring 
Committee (QASMC) semi-annually beginning six months after study activation at Washington 
University (if at least one patient has been enrolled) or one year after study activation (if no 
patients have been enrolled at the six-month mark). 
 
The DSM report for the DSMB will be prepared by the study team with assistance from the study 
statistician, will be reviewed by the DSMB, and will be submitted to the QASM Committee.  The 
DSMB must meet at least every six months beginning six months after enrollment of the first 
patient at a secondary site no more than one month prior to the due date of the DSM report to 
QASMC.  This report will include: 
• 
HRPO protocol number, protocol title, Principal Investigator name, data coordinator 
name, regulatory coordinator name, and statistician 
• 
Date of initial HRPO approval, date of most recent consent HRPO approval/revision, 
date of HRPO expiration, date of most recent QA audit, study status, and phase of 
study 
• 
History of study including summary of substantive amendments; summary of accrual 
suspensions including start/stop dates and reason; and summary of protocol 
exceptions, error, or breach of confidentiality including start/stop dates and reason 
• 
Study-wide target accrual and study-wide actual accrual including numbers from 
participating sites 
• 
Protocol activation date at each participating site 
• 
Average rate of accrual observed in year 1, year 2, and subsequent years at each 
participating site 
• 
Expected accrual end date, accrual by site, and accrual by cohort 
• 
Objectives of protocol with supporting data and list the number of participants who 
have met each objective 
• 
Measures of efficacy  
• 
Early stopping rules with supporting data and list the number of participants who have 
met the early stopping rules 
• 
Summary of toxicities at all participating sites and separated by cohorts with the 
number of dose-limiting toxicities indicated 
• 
Abstract submissions/publications 
• 
Summary of any recent literature that may affect the safety or ethics of the study 
 
Further DSMB responsibilities are described in the DSMB charter. 
 
The study principal investigator and coordinator will monitor for serious toxicities on an ongoing 
basis. Once the principal investigator or coordinator becomes aware of an adverse event, the AE 
will be reported to the HRPO and QASMC according to institutional guidelines (please refer to 
Section 7.0). 
 

Version 05/15/2023 
 
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Refer to the Washington University Quality Assurance and Data Safety Monitoring Committee 
Policies and Procedures for full details on the responsibilities of the DSMB. This is located on the 
QASMC website at https://siteman.wustl.edu/research/clinical-research-resources/protocol-
office-prmcqasmc/.   
 
 
13 
MEASUREMENT OF EFFECT 
 
13.1 
rhIL-7-hyFc effect 
 
The biological effect of rhIL-7-hyFc will be determined by its effect on raising ALC. ALC 
will be measured per the study calendar. The levels of ALC and the changes of ALC over 
time prior to and post rhIL-7-hyFc treatment will be monitored and analyzed (see statistical 
analysis section).  
 
13.2 
Antitumor Effect – RANO 
 
Brain MRI will be performed about every 2 months (±30 days) as per SOC to evaluate 
response. 
 
Criteria for response: Response will be evaluated in this study using the updated 
response assessment criteria for high-grade gliomas: Response Assessment in Neuro-
Oncology (RANO) working group guideline. 
 
Criteria for Response Assessment Incorporating MRI and Clinical Factors 
Response 
Criteria 
Complete response 
• 
Requires all of the following: complete disappearance of all enhancing 
measurable and nonmeasurable disease sustained for at least 4 weeks. 
• 
No new lesions; stable or improved nonenhancing (T2/FLAIR) lesions. 
• 
Patients must be off corticosteroids (or on physiologic replacement doses 
only) and stable or improved clinically. Note: Patients with nonmeasurable 
disease only cannot have a complete response; the best response possible is 
stable disease.  
Partial response 
Requires all of the following:  
• 
≥ 50% decrease compared with baseline in the sum of products of 
perpendicular diameters of all measurable enhancing lesions sustained for at 
least 4 weeks. 
• 
No progression of nonmeasurable disease. 
• 
Stable or improved nonenhancing (T2/FLAIR) lesions on same or lower dose 
of corticosteroids compared with baseline scan; the corticosteroid dose at the 
time of the scan evaluation should be no greater than the dose at time of 
baseline scan. 
• 
Stable or improved clinically. Note: Patients with nonmeasurable disease only 
cannot have a partial response; the best response possible is stable disease.  
Stable disease 
Requires all of the following:  
• 
Does not qualify for complete response, partial response, or progression. 
• 
Stable nonenhancing (T2/FLAIR) lesions on same or lower dose of 
corticosteroids compared with baseline scan. In the event that the 
corticosteroid dose was increased for new symptoms and signs without 

Version 05/15/2023 
 
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Response 
Criteria 
confirmation of disease progression on neuroimaging, and subsequent follow-
up imaging shows that this increase in corticosteroids was required because of 
disease progression, the last scan considered to show stable disease will be 
the scan obtained when the corticosteroid dose was equivalent to the baseline 
dose.  
Progression 
Defined by any of the following:  
• 
≥ 25% increase in sum of the products of perpendicular diameters of 
enhancing lesions compared with the smallest tumor measurement obtained 
either at baseline (if no decrease) or best response, on stable or increasing 
doses of corticosteroids*. The absolute increase in any dimension must be at 
least 5mm when calculating the products. 
• 
Significant increase in T2/FLAIR nonenhancing lesion on stable or increasing 
doses of corticosteroids compared with baseline scan or best response after 
initiation of therapy* not caused by comorbid events (e.g. radiation therapy, 
demyelination, ischemic injury, infection, seizures, postoperative changes, or 
other treatment effects). 
• 
Any new measureable lesion. 
• 
Clear clinical deterioration not attributable to other causes apart from the 
tumor (e.g. seizures, medication adverse effects, complications of therapy, 
cerebrovascular events, infection, and so on) or changes in corticosteroid 
dose. 
• 
Failure to return for evaluation as a result of death or deteriorating condition; 
or clear progression of nonmeasurable disease.  
• 
NOTE. All measurable and nonmeasurable lesions must be assessed using the same techniques 
as at baseline. 
• 
Abbreviations: MRI, magnetic resonance imaging; FLAIR, fluid-attenuated inversion recovery. 
• 
Stable doses of corticosteroids include patients not on corticosteroids. 
 
Criteria for progression: Pseudoprogression is a phenomenon where radiographic 
features are consistent with tumor recurrence/progression but is instead related to 
treatment effect. This is a common occurrence in glioblastoma following radiation that is 
generally seen in the first 3-6 months after completing therapy.  
 
If follow-up imaging confirms progression, the date of actual progression should be back-
dated to the date of initial radiographic evidence of progression. 
 
Alternatively, progressive disease can be defined as radiographic evidence of progression 
PLUS significant clinical decline that is felt to be unrelated to a co-morbid event or 
concurrent medication.   
  
13.3 
Disease Parameters 
 
Measurable disease:  Bi-dimensionally measurable lesions with clearly defined margins 
by MRI scan.  All tumor measurements must be recorded in millimeters (or decimal 
fractions of centimeters).   
 
Non-measurable or evaluable disease:  Uni-dimensionally measurable lesions or 
lesions with margins not clearly defined such as areas of T2/FLAIR signal abnormality or 
poorly defined enhancing abnormality.  
 
Note: For cystic lesions, the only measurable part is any enhancement area around the 

Version 05/15/2023 
 
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cyst that is clearly defined and bi-dimensionally measurable. The cyst itself should not be 
considered measurable or non-measureable disease. 
 
Target lesions: All measurable lesions that are residual of the lesion treated with MLA 
should be identified as target lesions and recorded and measured.  Target lesions should 
be selected on the basis of their size (lesions with the longest diameter), but in addition 
should be those that lend themselves to reproducible repeated measurements.  It may be 
the case that, on occasion, the largest lesion does not lend itself to reproducible 
measurement in which circumstance the next largest lesion, which can be measured 
reproducibly should be selected.  When there are too many measurable lesions, choose 
the largest 3 lesions as target lesions to follow.  The other measurable lesions should be 
considered evaluable for the purpose of objective status determination. 
 
Non-target lesions:  All non-measurable lesions should be identified as non-target 
lesions and should also be recorded at baseline.  Measurements of these lesions are not 
required, but the presence, absence, or in rare cases unequivocal progression of each 
should be noted throughout follow-up.  
 
13.4 
Methods for Evaluation of Measurable Disease 
 
All measurements should be taken and recorded in metric notation using a ruler.   
 
Clinical lesions:  Clinical lesions will only be considered measurable on brain MRI when 
they are ≥ 5 mm diameter as assessed using a ruler. 
 
Histology:  This technique can be used to differentiate between partial responses (PR) 
and complete responses (CR) in rare cases when biopsy or surgical resection of a 
measureable lesion is clinically indicated.  
 
Perfusion/CBV: This advanced brain MRI technique can be used as an adjunct test to 
determine treatment response or disease status.  However, it should not be used as the 
primary or sole method to determine response or disease status.  
 
13.4.1 Evaluation of Target Lesions 
 
Complete Response (CR):  Disappearance of all target lesions. 
 
Partial Response (PR):  ≥ 50% decrease compared with baseline in the sum of 
products of perpendicular diameters of all target lesions sustained for at least 4 
weeks. 
 
Progressive Disease (PD):  At least a 25% increase in the sum of products of 
perpendicular diameters of at least 1 target lesion, taking as reference the smallest 
sum of products of perpendicular diameters on study (this includes the baseline 
sum if that is the smallest on study).  The absolute increase in any dimension must 
be at least 5mm when calculating the products of perpendicular diameters. 
 
Stable Disease (SD):  Neither sufficient shrinkage to qualify for PR nor sufficient 
increase to qualify for PD, taking as reference the smallest sum of products of 
perpendicular diameters while on study. 
 

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13.4.2 Evaluation of Non-Target Lesions 
 
Complete Response (CR):  Disappearance of all non-target lesions.  
 
Non-CR/Non-PD:  Persistence of one or more non-target lesion(s). 
 
Progressive Disease (PD): Appearance of one or more new lesions and/or 
unequivocal progression of existing non-target lesions on stable or increasing 
doses of corticosteroids compared with baseline scan or best response after 
initiation of therapy not caused by comorbid events (e.g. radiation therapy, 
demyelination, ischemic injury, infection, seizures, postoperative changes, or other 
treatment effects). Unequivocal progression should not normally trump target 
lesion status.  It must be representative of overall disease status change, not a 
single lesion increase.     
 
Although a clear progression of “non-target” lesions only is exceptional, the opinion 
of the treating physician should prevail in such circumstances, and the progression 
status should be confirmed at a later time by the review panel (or Principal 
Investigator). 
 
13.4.3 Evaluation of Best Overall Response 
 
The best overall response is the best response recorded from the start of the 
treatment until disease progression/recurrence (taking as reference for 
progressive disease the smallest measurements recorded since the treatment 
started).  The patient's best response assignment will depend on the achievement 
of both measurement and confirmation criteria. 
 
Summary of the RANO Response Criteria (Adapted from [47]) 
Criterion 
CR 
PR 
SD 
PD 
T1 gadolinium enhancing 
disease 
None 
≥ 50% ↓ 
< 50% ↓ but < 25% ↑ 
≥ 25% ↑* 
T2/FLAIR 
Stable or ↓ Stable or ↓ Stable or ↓ 
↑* 
New lesion 
None 
None 
None 
Present* 
Corticosteroids 
None 
Stable or ↓ Stable or ↓ 
NA† 
Clinical status 
Stable or ↑ Stable or ↑ Stable or ↑ 
↓* 
Requirement for response All 
All 
All 
Any* 
Abbreviations: RANO, Response Assessment in Neuro-Oncology; CR, complete 
response; PR, partial response; SD, stable disease; PD, progressive disease; FLAIR, 
fluid-attenuated inversion recovery; NA, not applicable.  
* Progression occurs when this criterion is present.  
† Increase in corticosteroids alone will not be taken into account in determining 
progression in the absence of persistent clinical deterioration.  
 
13.4.4 Duration of Response 
 
Duration of overall response:  The duration of overall response is measured from 
the time measurement criteria are met for CR or PR (whichever is first recorded) 
until the first date that recurrent or progressive disease is objectively documented 

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(taking as reference for progressive disease the smallest measurements recorded 
since the treatment started). 
 
The duration of overall CR is measured from the time measurement criteria are 
first met for CR until the first date that progressive disease is objectively 
documented.  
 
Duration of stable disease:  Stable disease is measured from the start of the 
treatment until the criteria for progression are met, taking as reference the smallest 
measurements recorded since the treatment started, including the baseline 
measurements.  
 
13.4.5 Neurological Exam and Performance Status  
 
Patients will be graded using the Karnofsky Performance Status scale and their 
neurological function evaluated as improved, stable or deteriorated in addition to 
objective measurement of tumor size. These parameters will be used to determine 
the overall response assessment. 
 
13.4.6 Progression-Free Survival 
 
PFS is defined as the duration of time from start of treatment to time of progression 
or death, whichever occurs first. 
 
 
14 
AUDITING 
 
As coordinating center of this trial, Washington University (via the Quality Assurance and Safety 
Monitoring Committee (QASMC) will monitor each participating site to ensure that all protocol 
requirements are being met; that applicable federal regulations are being followed; and that best 
practices for patient safety and data collection are being followed per protocol.  Participating sites 
will be asked to send copies of all audit materials, including source documentation.  The audit 
notification will be sent to the Washington University Research Patient Coordinator, who will 
obtain the audit materials from the participating institution. 
 
Notification of an upcoming audit will be sent to the research team one month ahead of the audit. 
Once accrual numbers are confirmed, and approximately 30 days prior to the audit, a list of the 
cases selected for review (up to 10 for each site) will be sent to the research team. However, if 
during the audit the need arises to review cases not initially selected, the research team will be 
asked to provide the additional charts within two working days. 
Items to be evaluated include: 
• 
Subject screening and enrollment 
• 
Reporting of adverse events 
• 
Maintenance of HIPAA compliance 
• 
Completeness of regulatory documentation 
• 
Completeness of participant documentation 
• 
Acquisition of informed consent 
• 
IRB documentation 
• 
Issues of protocol adherence 
 

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Additional details regarding the auditing policies and procedures can be found at 
https://siteman.wustl.edu/research/clinical-research-resources/protocol-office-prmcqasmc/. 
 
 
15 
STATISTICAL CONSIDERATIONS 
  
15.1 
Study Design 
 
This is a single-center study consisting of a phase I study to identify MTD (/RP2D and a 
phase II placebo-controlled randomized study on target patients. In the phase I study, 
MTD will be determined and a recommended phase 2 dose (RP2D) will be decided. The 
maximum tolerated dose (MTD) is defined as the dose level immediately below the non-
tolerated dose. A total of at least 6 patients must be treated at a dose level for it to be 
considered the MTD. RP2D is the dose recommended for phase 2 study based on the 
MTD from dose escalation and ALC data which can never exceed MTD.  
  
During the randomized phase II portion of the study, eligible patients will be enrolled and 
randomized at a 1:1 ratio to either rhIL-7-hyFc at the RP2D dose level or placebo arm (see 
study Schema).  
 
Phase I enrollment was completed with N=19 patients.  The MTD was identified as 720 
mcg/kg.  Current collected data suggest an improvement in progression-free survival in 
patients with IDH1 wildtype as defined by immunohistochemistry negative staining using 
an IDH mutant-specific antibody and MGMT promoter unmethylated GBM; therefore, we 
have added an expansion cohort to the rhIL-7-hyFc arm with N=31 as justified in Section 
14.4. 
 
15.2 
The phase I study  
 
15.2.1 Study design 
 
The phase I dose finding study will begin with an Accelerated Phase with 1 patient 
per cohort at the first 2 doses (60 mcg/kg, 120 mcg/kg) followed by a conventional 
3+3 dose escalation phase on the remaining 4 doses (240 mcg/kg~960mcg/kg). A 
cohort of six patients need to be tested at the MTD/RP2D level. 
 
The Accelerated Phase 
The Safety data from current phase I studies on rhIL-7-hyFc in solid tumors and 
GBM show that patients tolerate either or both of the first 2 doses (60 mcg/kg, 120 
mcg/kg) well with no DLTs observed at the 2 doses, see rationale section on 
ABTC-1403 (IND 137067) on healthy volunteers, GX-I7-CA-003 in solid tumors, 
GX-I7-CA-004 in GBM. The Accelerated Phase will be conducted for dose levels 
1 to 2, i.e., dosage 60 mcg/kg and 120 mcg/kg, with 1 patient per dose level and 
DLT and adverse events will be monitored during the 1st cycle of the treatment. If 
no DLT is observed at dosage 60 mcg/kg, a new patient will be enrolled to dose 
level 2 (120 mcg/kg).  If any DLTs are observed in the 1 patient at either of the two 
dose levels, the Accelerated Phase will convert to a conventional 3+3 design at 
the dose level where DLTs are observed and remain so for all the dose levels 
above. If excess DLTs (>=2 out of a total of 6 patients experiencing DLTs) are 
observed at dose level 1 (60 mcg/kg), the study will halt. If no DLTs, the phase I 

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study will proceed to the next conventional 3+3 phase for dose levels 3 to 6 (see 
dosage Table in Section 5.2). 
 
The conventional 3+3 Phase 
Assuming no conversion at the first 2 doses, the conventional 3+3 design with 3 
independent patients per cohort will start at dose level 3 (240 mcg/kg) and will 
follow the dose escalation decision rule of 3+3 design to escalate along the 
dosages from dose level 3 (240 mcg/kg) to 6 (960 mcg/kg) sequentially from the 
lowest to the highest dose level, see dosage Table in Section 5.2. The dose 
escalation rules are summarized in the dose escalation criteria in Section 5.3.3. If 
excess DLT (≥2 out of a cohort of 3 patients or ≥2 out of a total of 6 patients from 
2 cohorts) is observed at starting dose level 240 mcg/kg, a cohort of 5 patients 
(six, plus the one patient tested previously) will be tested at dose level 2 (120 
mcg/kg) and if <=1 DLTs in 6 are observed, dose level 2 will be the recommended 
dose for the Phase II. If >=2 out of 6 patients are observed with DLTs at dose level 
2, the trial will similarly test dose level 1 to have dose level 1 either as the MTD or 
the study halt in presence of excess toxicities. If no excess dose (0 DLT out of a 
cohort of 3 patients or 1 DLT out of a total of 6 patients) is observed at dose level 
3, the Phase I study will follow the dose escalation decision rule to escalate along 
the dosages thereafter to dose level 6 until the MTD if no DLT at highest does 
level) is identified at which no more than one in six patients experiences DLT. A 
total of 6 patients will need to be treated at the MTD level. Integrating the MTD 
data and ALC data, a RP2D (which can be the MTD or not but will not exceed the 
MTD), will be determined.  
 
If we do not find MTD at the proposed dose levels, we plan to amend the protocol 
to test new or higher dose levels per the recommendations from NIT. In addition, 
another Phase I trial is on-going. Dose escalation can be stopped if a MTD is 
determined through the on-going Phase I trial announced by the sponsor.  
 
15.2.2 Endpoint 
 
The primary endpoint of the phase I is adverse events (AEs), especially dose 
limiting toxicity (DLT). DLT will be defined as ≥ grade 3 AEs that occur within 30 
days from the date when patients receive the 1st dose of rhIL-7-hyFc administration 
and are concluded to be possibly, likely or definitely related to the drug regimen 
that occurs during cycle 1, with severity graded according to the Common 
Terminology Criteria for Adverse Events (CTCAE) 5.0.  Grade 3 dermatologic AEs, 
including injection site reaction, that resolves to Grade ≤ 2 in ≤ 3 days with 
supportive care; transient Grade 3 influenza-like symptoms or pyrexia controlled 
with medical management; Grade 3 fatigue that resolves to Grade ≤.2 in ≤ 3days; 
Grade 3 neurologic toxicity responding within ≤ 7 days to steroids, anticonvulsants, 
or electrolyte correction; or Grade 3 laboratory abnormalities that are 
asymptomatic and considered by the investigator not to be clinically significant will 
be excluded from the DLT definition. 
The secondary endpoint is absolute lymphocyte count (ALC). 
 
 
 
 

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15.2.3 Sample size calculation 
 
During the Phase I, total number of patients enrolled will depend on the number of 
dose levels and observed DLTs, ranging from a minimal of 6 patients to a 
maximum of 26 patients.  
 
15.2.4 Statistical Analysis 
 
Data from the Phase I study will be presented in a descriptive manner AEs and 
SAEs will be summarized by counts and percentages by patient, dose type and 
grade.   
 
15.3 
The randomized phase II study 
 
15.3.1 Study design 
 
For the phase II trial, eligible patients, stratified by concomitant use of steroids 
(Yes/No), will be randomized at a 1:1 ratio to receive either placebo or the study 
drug rhIL-7-hyFc. Patients on the study drug arm will receive multiple doses of the 
study drug rhIL-7-hyFc (see study schema). Lymphocyte count will be measured 
before and after rhIL-7-hyFc administration (details see treatment schema and 
study calendar). 
 
15.3.2 Endpoint 
 
The primary endpoint of Phase II is the % increase of ALC in evaluable patients 
prior to adjuvant TMZ (at ~ Week 4) that measures the “pure” drug effect before 
influences from TMZ in reference to the baseline prior to rhIL-7-hyFc injection. A 
patient is considered evaluable for the primary endpoint if (s)he completes CBC at 
baseline and at ~4 weeks after the first dose of NT-17 or placebo and has valid 
ALC measurements prior to and at ~week 4 before adjuvant TMZ of treatment with 
NT-17 or placebo. Enrolled patients who are not evaluable for the primary endpoint 
will be replaced so as to meet the study power requirements (see 15.3.3). 
 
 The secondary endpoints include lymphocyte counts over time in the setting of 
steroid use and chemotherapy, progression free survival (PFS) and overall survival 
(OS). 
 
15.3.3 Sample size calculation 
 
We summarized the ALC of all comers seen in our center at 1 month after 
completion of RT + TMZ and estimated a mean of ALC at 850 and a standard 
deviation (SD) of 480. This also agrees with data in published literatures (Mendez 
2016).  We assume that patients on the placebo arm (who usually have received 
radiation and TMZ post-operative) have the same mean of 850 and SD of 480. 
Based on 1-sided two sample t-test with equal variance at a 5% level, 8 patients 
per arm provides 80% power to test a mean ALC of 1486 in the treated patients in 
comparison to a mean of 850 in the placebo arm. Assuming ~10% dropout rate, 
we will enroll a total of 20 patients, 10 per arm.  
 

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For the whole study spanning through the Phase I and the Phase II study, 
approximately a minimum of 26 to a maximum of 46 evaluable patients will be 
enrolled.  
 
15.3.4 Accrual  
 
We see approximately 150 new patients with HGG (with ~70% are GBM) each 
year at Washington University with ~30% participating in clinical trials. We expect 
to be able to enroll approximately 1-2 patient per month on average, and thus it 
will take approximately 18 months to enroll ~29 patients and 30 months to enroll 
the expected maximum total of 46 evaluable patients. 
 
15.3.5 Statistical Analysis 
 
All data will be analyzed as observed and no imputation for missing values will be 
used. Patient characteristics will be summarized by descriptive statistics.  
Descriptive statistics will be used to summarize the trial results, i.e., statistics for 
continuous variables such as lymphocyte counts may include means, medians, 
ranges and appropriate measures of variability. Qualitative variables will be 
summarized by counts and percentages. The uncertainty of estimates will be 
assessed by confidence intervals (CIs). Markers for lymphocyte recovery in each 
group will be summarized as means and standard deviations (SD) at each time 
point.  The primary endpoint, ALC at ~ week 4 and the change from the time point 
of radiation will be compared between the two arms by two sample t-test or 
Wilcoxon rank sum test as appropriate and by a linear model where other important 
factors such as concomitant use of steroid besides the treatment arm will be 
modeled. Within each arm, the primary endpoint will be compared to the ALC at 
the completion of radiation by paired sample t-test or Wilcoxon signed rank test as 
proper. Total lymphocyte counts at later cycles will be similarly analyzed but may 
be compared to a different reference time point.  We will analyze ALC over time 
using linear models for repeated measurement data, followed by post-hoc multiple 
comparisons for between-group differences at other time points of interest. PFS 
and overall survival will be analyzed using the Kaplan–Meier method and survival 
difference will be compared between arms by log rank test. The univariate Cox 
proportional hazard model will be applied to evaluate ALC at each time point, in 
continuous scale and dichotomized scale by the cutoff of 1 (determined based on 
literature), for their prognostic effect on PFS and OS while multivariate Cox model 
will be applied to account for other covariates. The time varying Cox proportional 
hazard model will be applied to model the prognostic effect of the longitudinal ALC 
as a whole, without and with additional covariates. 
 
15.4 
The expansion cohort 
  
15.4.1 Study design 
 
Based on observed survival improvement in currently enrolled patients, we added 
(in August 2021) an expansion cohort to the experimental arm. For the expansion 
cohort, a total of N=31 IDH1 wildtype confirmed by immunohistochemistry and 
MGMT promoter unmethylated eligible patients will be enrolled to receive the study 
drug rhIL-7-hyFc.  
 

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15.4.2 Endpoint 
 
The primary endpoint for the expansion cohort is progression free survival (PFS) 
defined from date of surgery to date of progression or death due to disease or date 
of last clinical follow up.  The secondary endpoints include AE and OS. 
 
15.4.3 Sample size calculation 
 
The sample size calculation was based on testing the estimated median PFS of 
11.5 months in the currently enrolled patients versus a null median PFS of 6.3 
months (NEJM, 2014, 8:370), assuming PFS follows an exponential distribution. 
 
A two-sided, one-sample log-rank test calculated from N=31 subjects achieves 
81.05% power at a 5% significance level to detect a median survival time of 11.5 
months (see Section 1.8.2) when the median survival time of the historic control 
group is 6.3 months, corresponding to a hazard ratio of 0.548. Subjects are 
accrued for a period of 18 months. Follow-up continues for a period of 12 months 
after the last subject is added. The probability that a subject experiences an event 
during the study is 0.7039. The expected number of events during the study with 
N=31 eligible patients is 22. 
  
15.4.4 Accrual  
 
We see approximately 150 new patients with HGG (with ~70% are GBM) each 
year at Washington University with ~30% participating in clinical trials. We expect 
to be able to enroll approximately 1-2patient per month on average, and thus we 
can enroll 31 patients within approximately 18-month accrual period. 
 
15.4.5 Statistical Analysis 
 
PFS will be analyzed by the KM method and median PFS will be estimated with 
95% CI and tested against the null median PFS of 6.3 months by 2-sided one-
sample log rank test. OS will be analyzed similarly. Other endpoints will be 
analyzed similarly as in Section 15.3.5 but without between arm comparison.  
 
15.5 
Analysis set 
 
The safety analysis set include all patients who have received any dose of the rhIL-7-hyFc. 
 
The efficacy analysis set for the randomized phase II portion of the study includes all 
patients who have received a dose of rhIL-7-hyFc, completes CBC at baseline and at ~4 
weeks prior to adjuvant TMZ, and have evaluable ALC measurements from the CBC labs. 
The primary analyses will focus on the intent to treat (ITT) patients regardless what 
treatment they actually receive. 
 
The PFS evaluable patients include patients who received at least one dose of rhIL-7-
hyFc.  
 
 

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15.6 
Early stopping rules 
 
The study will stop if the following stopping rules are met: 
 
In phase I part of the study, if ≥2 patients are observed with DLTs out of 6 patients at dose 
level 1 (60 mcg/kg), the trial will halt.  
 
 
There is no efficacy stopping rule for the randomized phase II cohort or expansion cohort. 
The toxicity data for the expansion cohort will be reviewed regularly by the study team and 
the DSMB. If serious unexpected toxicities occur, the expansion cohort can be stopped 
early due to safety concern as recommended by the DSMB. 
 
15.7 
Ad hoc proof-of-concept check 
In the randomized phase II portion of the study, when the number of patients enrolled 
into each arm is 5 (half of N per arm), we will perform one-time ad-hoc proof-of-concept 
check on the primary endpoint, the primary % ALC change at ~wk4 prior to adjuvant 
TMZ relative to baseline . If the ad-hoc check finds that the mean % ALC increase 
evaluated in the rhlL-7-hyFc treated patients is lower than that of placebo treated 
patients, the study will halt to be reviewed by the PI and DSMB. 
 
16 
MULTICENTER REGULATORY REQUIREMENTS 
 
Washington University requires that each participating site sends its informed consent document 
to be reviewed and approved by the Washington University Regulatory Coordinator (or designee) 
prior to IRB/IEC submission.    
 
Site activation is defined as when the secondary site has received official written documentation 
from the coordinating center that the site has been approved to begin enrollment.  At a minimum, 
each participating institution must have the following documents on file at Washington University 
prior to study activation: 
• 
Documentation of IRB approval of the study in the form of a letter or other official 
document from the participating institution’s IRB.  This documentation must show 
which version of the protocol was approved by the IRB. 
• 
Documentation of IRB approval of an informed consent form. The consent must 
include a statement that data will be shared with Washington University, including the 
Quality Assurance and Safety Monitoring Committee (QASMC), the DSMB (if 
applicable), the Washington University study team, and NeoImmuneTech. 
• 
Documentation of FWA, signed FDA Form 1572 (if applicable), and the CVs of all 
participating investigators. 
• 
Protocol signature page signed and dated by the investigator at each participating site. 
 
The coordinating center Principal Investigator (or designee) is responsible for disseminating to 
the participating sites all study updates, amendments, reportable adverse events, etc.  
Protocol/consent modifications and IB updates will be forwarded electronically to the secondary 
sites within 4 weeks of obtaining Washington University IRB approval.  Activated secondary sites 
are expected to submit protocol/consent/IB modifications to their local IRBs within 4 weeks of 
receipt unless otherwise noted.  Upon the secondary sites obtaining local IRB approval, 

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documentation of such shall be sent to the Washington University study team within 2 weeks of 
receipt of approval. 
 
Documentation of participating sites’ IRB approval of annual continuing reviews, protocol 
amendments or revisions, all SAE reports, and all protocol violations/deviations/exceptions must 
be kept on file at Washington University. 
 
The investigator or a designee from each institution must participate in a regular conference call 
to update and inform regarding the progress of the trial. 
 
 

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17 
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APPENDIX A: NANO Scale 
 
 
 
 

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APPENDIX B: Safety Data 
 
In Study GX-I7-CA-003 of single-agent rhIL-7-hyFc given in multiple doses in advanced solid 
tumor patients, there were no DLTs observed up to 1200ug/kg. Each of the cohorts (Cohort 1: 60 
ug/kg; Cohort 2: 120ug/kg; Cohort 3: 240ug/kg; Cohort 4: 480ug/kg; Cohort 5: 720ug/kg; Cohort 
6: 960ug/kg; Cohort 7: 1200ug/kg) consisted of 3 patients each that followed a 3+3 dose 
escalation design. 
 
Following are the patient baseline characteristics: 
 
 
 
 
 
 

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The most common AEs seen in the study were injection site reactions (Grades 1 or 2 only). 
Following is a list of AEs seen in each of the cohorts. 
 
 
 
 
 
 
 

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APPENDIX C: Definitions for Adverse Event Reporting 
 
A. Adverse Events (AEs) 
 
As defined in 21 CFR 312.32: 
 
Definition: any untoward medical occurrence associated with the use of a drug in humans, 
whether or not considered drug-related. 
 
Grading: the descriptions and grading scales found in the revised NCI Common Terminology 
Criteria for Adverse Events (CTCAE) version 5.0 will be utilized for all toxicity reporting.  A 
copy of the CTCAE version 5.0 can be downloaded from the CTEP website. 
 
Attribution (relatedness), Expectedness, and Seriousness: the definitions for the terms 
listed that should be used are those provided by the Department of Health and Human 
Services’ Office for Human Research Protections (OHRP).  A copy of this guidance can be 
found on OHRP’s website: 
http://www.hhs.gov/ohrp/policy/advevntguid.html 
 
B. Suspected Adverse Reaction (SAR) 
 
As defined in 21 CFR 312.32: 
 
Definition: any adverse event for which there is a reasonable possibility that the drug caused 
the adverse event.  “Reasonable possibility” means there is evidence to suggest a causal 
relationship between the drug and the adverse event.  “Suspected adverse reaction” implies 
a lesser degree of certainty about causality than adverse reaction, which means any adverse 
event caused by a drug. 
 
C. Life-Threatening Adverse Event / Life Threatening Suspected Adverse Reaction  
 
As defined in 21 CFR 312.32: 
 
Definition: any adverse drug event or suspected adverse reaction is considered “life-
threatening” if, in the view of the investigator, its occurrence places the patient at immediate 
risk of death. It does not include an adverse event or suspected adverse reaction that, had it 
occurred in a more severe form, might have caused death. 
 
D.  Serious Adverse Event (SAE) or Serious Suspected Adverse Reaction 
 
As defined in 21 CFR 312.32: 
 
Definition:  an adverse event or suspected adverse reaction is considered “serious” if, in the 
view of the investigator, it results in any of the following outcomes: 
o Death 
o A life-threatening adverse event 
o Inpatient hospitalization or prolongation of existing hospitalization 
o A persistent or significant incapacity or substantial disruption of the ability to conduct 
normal life functions 
o A congenital anomaly/birth defect 

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o Any other important medical event that does not fit the criteria above but, based upon 
appropriate medical judgment, may jeopardize the subject and may require medical or 
surgical intervention to prevent one of the outcomes listed above 
 
E. Protocol Exceptions 
 
Definition: A planned change in the conduct of the research for one participant. 
 
F. Deviation 
 
Definition: Any alteration or modification to the IRB-approved research without prospective 
IRB approval.  The term “research” encompasses all IRB-approved materials and documents 
including the detailed protocol, IRB application, consent form, recruitment materials, 
questionnaires/data collection forms, and any other information relating to the research study. 
 
A minor or administrative deviation is one that does not have the potential to negatively impact 
the rights, safety, or welfare of participants or others or the scientific validity of the study. 
 
A major deviation is one that does have the potential to negatively impact the rights, safety, 
or welfare of participants or others or the scientific validity of the study. 
 

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APPENDIX D: Reporting Timelines 
 
Expedited Reporting Timelines 
Event 
HRPO 
QASMC 
FDA 
Drug/Device Manufacturer 
Serious AND unexpected 
suspected adverse reaction 
 
 
Report no later than 15 
calendar days after it is 
determined that the 
information qualifies for 
reporting 
All events being submitted to 
the FDA should be submitted 
concurrently to NeoImmune 
Tech 
Unexpected fatal or life-
threatening suspected adverse 
reaction 
 
 
Report no later than 7 calendar 
days after initial receipt of the 
information 
All events being submitted to 
the FDA should be submitted 
concurrently to NeoImmune 
Tech 
Unanticipated problem 
involving risk to participants or 
others 
Report within 10 working days.  
If the event results in the death 
of a participant enrolled at 
WU/BJH/SLCH, report within 1 
working day. 
Report via email after IRB 
acknowledgment 
 
All events being submitted to 
the QASMC should be 
submitted concurrently to 
NeoImmuneTech 
Major deviation 
Report within 10 working days.  
If the event results in the death 
of a participant enrolled at 
WU/BJH/SLCH, report within 1 
working day. 
 
 
All events being submitted to 
the HRPO should be submitted 
concurrently to 
NeoImmuneTech 
A series of minor deviations 
that are being reported as a 
continuing noncompliance 
Report within 10 working days.    
 
 
Protocol exception 
Approval must be obtained 
prior to implementing the 
change 
 
 
 
Clinically important increase in 
the rate of a serious suspected 
adverse reaction of that list in 
the protocol or IB 
 
 
Report no later than 15 
calendar days after it is 
determined that the 
information qualifies for 
reporting 
All events being submitted to 
the FDA should be submitted 
concurrently to 
NeoImmuneTech 
Complaints 
If the complaint reveals an 
unanticipated problem 
involving risks to participants 
or others OR noncompliance, 
report within 10 working days.  
 
 
 

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Expedited Reporting Timelines 
Event 
HRPO 
QASMC 
FDA 
Drug/Device Manufacturer 
If the event results in the death 
of a participant enrolled at 
WU/BJH/SLCH, report within 1 
working day.  Otherwise, 
report at the time of continuing 
review. 
Breach of confidentiality 
Within 10 working days. 
 
 
 
Incarceration 
If withdrawing the participant 
poses a safety issue, report 
within 10 working days.   
 
If withdrawing the participant 
does not represent a safety 
issue and the patient will be 
withdrawn, report at continuing 
review. 
 
 
 
 
Routine Reporting Timelines 
Event 
HRPO 
QASMC 
FDA 
Drug/Device Manufacturer 
Adverse event or SAE that 
does not require expedited 
reporting 
If they do not meet the 
definition of an unanticipated 
problem involving risks to 
participants or others, report 
summary information at the 
time of continuing review 
Adverse events will be 
reported in the toxicity table in 
the DSM report which is 
typically due every 6 months. 
The most current toxicity table 
from the DSM report is 
provided to the FDA with the 
IND’s annual report. 
All events being submitted to 
the FDA should be submitted 
concurrently to 
NeoImmuneTech 
Minor deviation 
Report summary information at 
the time of continuing review. 
 
 
 
Complaints 
If the complaint reveals an 
unanticipated problem 
involving risks to participants 
or others OR noncompliance, 
report within 10 working days.  
If the event results in the death 
of a participant enrolled at 
WU/BJH/SLCH, report within 1 
working day.  Otherwise, 
report at the time of continuing 
review. 
 
 
 

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Incarceration 
If withdrawing the participant 
poses a safety issue, report 
within 10 working days.   
 
If withdrawing the participant 
does not represent a safety 
issue and the patient will be 
withdrawn, report at continuing 
review. 
 
 
 
 
 
Expedited Reporting Timelines for Secondary Sites 
Event 
WU (Coordinating Center) 
Local IRB 
FDA 
Drug/Device Manufacturer 
Serious AND unexpected 
suspected adverse reaction 
Report no later than 11 calendar days 
after it is determined that the information 
qualifies for reporting. 
Report all applicable 
events to local IRB 
according to local 
institutional guidelines. 
The research team at 
Washington University is 
responsible for reporting all 
applicable events to the 
FDA as needed. 
The research team at 
Washington University is 
responsible for reporting all 
applicable events to 
NeoImmuneTech as needed. 
Unexpected fatal or life-
threatening suspected 
adverse reaction 
Report no later than 4 calendar days after 
initial receipt of the information. 
Unanticipated problem 
involving risk to participants 
or others 
Report no later than 4 calendar days after 
initial receipt of the information. 
Adverse event or SAE that 
does not require expedited 
reporting 
As per routine data entry expectations 
Protocol exception 
Approval must be obtained prior to 
implementing the change. 
 
 
 
 
 
 
 
 
 
 
 

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APPENDIX E: Washington University SAE Reporting Cover Sheet 
 
SAE COVER SHEET- Secondary Site Assessment 
 
Washington University HRPO#:  
Sponsor-Investigator:  
Subject Initials:   
Subject ID:  
Treating MD:  
Treating Site:  
EVENT TERM:  
Event Start Date:  
EVENT GRADE: 
Date of site’s first notification: 
 
Treating MD Event Assessment:  
 
 
Is this event possibly, probably, or definitely related study treatment? 
 
 yes  
 
 no 
 
If yes, please list which drug (if more than one)______________________________ 
 
       
Explain 
______________________________________________________________________ 
 
____________________________________________________________________________ 
 
 
_______________________           ___________________________     _________________  
Physician’s Name 
 
 
Physician’s Signature 
 
Date