A Phase 1 Clinical Trial to Evaluate the Safety and Immunogenicity of a Neoantigen DNA Vaccine Strategy in Pancreatic Cancer Patients Following Surgical Resection and Adjuvant Chemotherapy
Phase: PHASE1
Conditions: Pancreatic Cancer, Pancreas Cancer, Cancer of the Pancreas
Sponsor: Washington University School of Medicine

Washington University School of Medicine 
Protocol Date 08/03/23, A5 
Siteman Cancer Center 
Pancreatic Cancer SPORE 
Neoantigen DNA vaccines  
 
Page 1 of 69 
A PHASE 1 CLINICAL TRIAL TO EVALUATE THE SAFETY AND IMMUNOGENICITY 
OF A NEOANTIGEN DNA VACCINE STRATEGY IN PANCREATIC CANCER 
PATIENTS FOLLOWING SURGICAL RESECTION AND ADJUVANT 
CHEMOTHERAPY 
HRPO #: 201708105 
Version date: 08/03/2023 
IND #: 17596 
ClinicalTrials.gov #: NCT03122106 
 
INVESTIGATORS 
Principal Investigators: 
Daniel A. Laheru, MD 
Medical Oncologist 
Johns Hopkins School of Medicine 
401 N. Broadway 
Baltimore, MD 21231 
Telephone: 410-955-8974 
E-mail: Laherda@jhmi.edu 
 
William E. Gillanders, M.D. 
IND Sponsor, Surgical Oncologist 
Washington University School of Medicine 
660 South Euclid, Box 8109 
St. Louis, MO 63110 
Telephone: 314-747-0072 
E-mail: gillandersw@wustl.edu 
 
Co-Investigators: 
Robert D. Schreiber, Ph.D. 
Immunobiologist 
Washington University School of Medicine 
660 South Euclid, Box 8118 
St. Louis, MO 63110 
Telephone: 314-362-8747 
E-mail: Schreiber@wustl.edu  
 
Christopher A. Miller, Ph.D. 
Genome Scientist 
The Genome Center at WUSM 
660 South Euclid, Box 8501 
St. Louis, MO 63110 
Telephone: 314-286-0263 
E-mail: c.a.miller@wustl.edu 
 
Elizabeth Jaffee, M.D. 
Medical Oncologist 
Johns Hopkins School of Medicine 
1650 Orleans Street, Cancer Research Bldg #4M07 
Baltimore, MD 21231 
Telephone: 410-614-8216 
E-mail: ejaffee@jhmi.edu 
 
Feng Gao, M.D., Ph.D. 
Biostatistician 
Washington University School of Medicine 
660 South Euclid, Box 8067 
St. Louis, MO 63110 
Telephone: 314-362-3682 
E-mail: feng@wustl.edu 
 
S. Peter Goedegebuure, Ph.D. 
Immunologist 
Washington University School of Medicine 
660 South Euclid, Box 8109 
St. Louis, MO 63110 
Telephone: 314-362-8612 
E-mail: goedegep@wustl.edu 
 
William Hawkins, M.D. 
Surgical Oncologist 
Washington University School of Medicine 
660 South Euclid, Box 8109 
St. Louis, MO 63110 
Telephone: 314-362-7046 
E-mail: hawkinsw@wudosis.wustl.edu 
 
Kian H. Lim, MD 
Medical Oncologist 
Washington University School of Medicine 
660 South Euclid, Box 8089 
St. Louis, MO 63110 
Telephone: 314-747-9320 
E-mail: klim@dom.wustl.edu 
 
Marianna Ruzinova, M.D., Ph.D. 
Pathologist 
Washington University School of Medicine 
BJCIH 03303 
St. Louis, MO 63110 
Telephone: 314-362-7704 
E-mail: mruzinova@wustl.edu  

Washington University School of Medicine 
Protocol Date 08/03/23, A5 
Siteman Cancer Center 
Pancreatic Cancer SPORE 
Neoantigen DNA vaccines  
 
Page 2 of 69 
SUMMARY 
Protocol: 
A PHASE 1 CLINICAL TRIAL TO EVALUATE THE SAFETY AND 
IMMUNOGENICITY OF A NEOANTIGEN DNA VACCINE STRATEGY IN 
PANCREATIC CANCER PATIENTS FOLLOWING SURGICAL RESECTION 
AND ADJUVANT CHEMOTHERAPY 
Study Design: 
This is a phase 1 open-label study to evaluate the safety and immunogenicity of 
a neoantigen DNA vaccine strategy in pancreatic cancer patients following 
surgical resection and adjuvant chemotherapy. The neoantigen DNA vaccines 
will incorporate prioritized neoantigens and personalized mesothelin epitopes 
and will be administered with an electroporation device. The hypothesis of this 
study is that neoantigen DNA vaccines will be safe and capable of generating 
measurable neoantigen-specific CD4 and CD8 T cell responses. The primary 
objective of this study is to demonstrate the safety and feasibility of the 
neoantigen DNA vaccine strategy. The secondary objective is to evaluate the 
immunogenicity of the neoantigen DNA vaccine strategy as measured by 
ELISPOT analysis, a surrogates for T cell function. 
Product Description: The neoantigen DNA vaccines are composed of closed circular DNA plasmids 
that are designed to express mutant tumor specific antigens identified by exome 
sequencing. The plasmids will be formulated as naked DNA plasmid vaccines. 
Vaccine vials will be supplied at a concentration of 2 mg/mL. DNA vaccines will 
be administered intramuscularly (in the deltoid or lateralis) using a TriGrid 
electroporation device (Ichor Medical Systems). Two injections (2 mg/injection, 
total dose 4 mg) will be administered at each time point. 
Subjects: 
Pancreatic cancer patients who have completed surgical resection and adjuvant 
chemotherapy without evidence of recurrent disease are eligible. Patients 
enrolled into the protocol must provide consent for exome sequencing and 
dbGAP-based data sharing and provide germline and tumor DNA samples of 
adequate quality for sequencing. 
Study Plan: 
Fifteen pancreatic cancer patients will be enrolled. Subjects will be treated by 
electroporation with 4 mg of a neoantigen DNA vaccine at weeks 1, 5, 9, 13, 17, 
and 21 ± 7 days with at least 21 days between injection days. Each DNA 
vaccination will be 4 mg vaccine administered intramuscularly using a TriGrid 
electroporation device. 
Study Duration: 
Each subject will be followed for 12 months following the last vaccination. 
Additional follow-up visits or telephone contact will be scheduled annually 
thereafter if the patient is alive and available for follow-up. 
Study Endpoints: 
The primary endpoint is safety of the neoantigen DNA vaccine regimen. Safety 
will be closely monitored after injection with eight or more clinical and laboratory 
assessments in the first six months of the trial. Toxicity will be graded according 
to the National Cancer Institute Common Terminology Criteria for Adverse 
Events version 4.0. The secondary endpoint is immunogenicity of the vaccine 
regimen as measured by ELISPOT analyses. Exploratory analyses will include 
other measures of the immunogenicity of the vaccine regimen as measured by 
multiparametric flow cytometry and other measures of immune function, including 
CYTOF analysis. 
 

Washington University School of Medicine 
Protocol Date 08/03/23, A5 
Siteman Cancer Center 
Pancreatic Cancer SPORE 
Neoantigen DNA vaccines  
 
Page 3 of 69 
SCHEMA 
 
 
 

Washington University School of Medicine 
Protocol Date 08/03/23, A5 
Siteman Cancer Center 
Pancreatic Cancer SPORE 
Neoantigen DNA vaccines  
 
Page 4 of 69 
PROTOCOL SIGNATURE PAGE 
 
A PHASE 1 CLINICAL TRIAL TO EVALUATE THE SAFETY AND IMMUNOGENICITY 
OF A NEOANTIGEN DNA VACCINE STRATEGY IN PANCREATIC CANCER 
PATIENTS FOLLOWING SURGICAL RESECTION AND ADJUVANT 
CHEMOTHERAPY 
 
I have read the attached clinical protocol and agree to conduct this trial in accordance with all the 
stipulations of the protocol and in accordance with the Declaration of Helsinki/Tokyo/Venice on 
Experimentation in Humans as required by the United States Food and Drug Administration regulations, 
Code of Federal Regulations Title 21 parts 50, 56, 312, 800, Title 45 part 46 and all applicable guidelines. 
 
Name of Investigator: 
 
 
Signature 
Date 
 

Washington University School of Medicine 
Protocol Date 08/03/23, A5 
Siteman Cancer Center 
Pancreatic Cancer SPORE 
Neoantigen DNA vaccines  
 
Page 5 of 69 
TABLE OF CONTENTS 
SUMMARY _________________________________________________________________________ 2 
SCHEMA ___________________________________________________________________________ 3 
PROTOCOL SIGNATURE PAGE _______________________________________________________ 4 
1 
OBJECTIVES ___________________________________________________________________ 7 
 
PRIMARY OBJECTIVE _________________________________________________________ 7 
 
SECONDARY OBJECTIVE _______________________________________________________ 7 
 
EXPLORATORY OBJECTIVES ____________________________________________________ 7 
2 
BACKGROUND _________________________________________________________________ 7 
 
PANCREATIC CANCER _________________________________________________________ 7 
 
MUTATIONAL LANDSCAPE IN PANCREATIC CANCER ____________________________________ 7 
 
TUMOR MICROENVIRONMENT ___________________________________________________ 7 
 
TUMOR ANTIGENS ___________________________________________________________ 8 
 
NEXT GENERATION SEQUENCING ________________________________________________ 8 
 
PERSONALIZED CANCER VACCINES _______________________________________________ 9 
 
PRIORITIZATION OF SEQUENCING-IDENTIFIED NEOANTIGENS _____________________________ 9 
 
POLYEPITOPE DNA VACCINES __________________________________________________ 9 
 
ELECTROPORATION _________________________________________________________ 10 
 
CLINICAL TRIAL DESIGN AND DOSE CONSIDERATIONS _________________________________ 10 
 
CHALLENGES OF SEQUENCING PANCREATIC CANCERS ________________________________ 11 
3 
PATIENT SELECTION __________________________________________________________ 11 
 
INCLUSION CRITERIA _________________________________________________________ 11 
 
EXCLUSION CRITERIA ________________________________________________________ 12 
 
INCLUSION OF WOMEN AND MINORITIES ___________________________________________ 13 
4 
REGISTRATION PROCEDURES __________________________________________________ 13 
 
PRIOR TO REGISTRATION _____________________________________________________ 13 
 
REGISTRATION PROCEDURES __________________________________________________ 14 
 
CONFIRMATION OF PATIENT ELIGIBILITY ___________________________________________ 15 
 
PATIENT REGISTRATION IN THE SITEMAN CANCER CENTER ONCORE DATABASE _____________ 15 
 
ASSIGNMENT OF UPN _______________________________________________________ 15 
5 
INVESTIGATIONAL AGENT ______________________________________________________ 15 
 
CHEMICAL NAME AND STRUCTURE _______________________________________________ 15 
 
NONCLINICAL STUDIES _______________________________________________________ 17 
 
SEQUENCING PIPELINE _______________________________________________________ 20 
 
EPITOPE PREDICTION ALGORITHM _______________________________________________ 21 
 
MANUFACTURING FACILITY ____________________________________________________ 22 
 
MANUFACTURING PROCESS ___________________________________________________ 24 
 
PRODUCT FORMULATION _____________________________________________________ 27 
 
PRODUCT RELEASE TESTS ____________________________________________________ 28 
 
STABILITY ________________________________________________________________ 29 
 
LABELS __________________________________________________________________ 29 
 
INVESTIGATIONAL AGENT ADMINISTRATION ________________________________________ 29 
 
INVESTIGATIONAL AGENT ACCOUNTABILITY ________________________________________ 33 
6 
TREATMENT PLAN_____________________________________________________________ 33 
 
PLASMID DNA ADMINISTRATION ________________________________________________ 33 
 
DEFINITIONS OF EVALUABILITY _________________________________________________ 34 
 
DURATION OF THERAPY ______________________________________________________ 34 
 
DURATION OF FOLLOW UP ____________________________________________________ 34 

Washington University School of Medicine 
Protocol Date 08/03/23, A5 
Siteman Cancer Center 
Pancreatic Cancer SPORE 
Neoantigen DNA vaccines  
 
Page 6 of 69 
7 
POTENTIAL TOXICITY AND DOSE MODIFICATIONS _________________________________ 35 
 
PREVIOUS HUMAN EXPERIENCE_________________________________________________ 35 
 
POTENTIAL TOXICITY ________________________________________________________ 36 
 
TOXICITY MONITORING AND MANAGEMENT _________________________________________ 38 
 
DOSE MODIFICATIONS _______________________________________________________ 38 
8 
STUDY CALENDAR ____________________________________________________________ 39 
9 
CRITERIA FOR RESPONSE ______________________________________________________ 40 
 
PRIMARY OBJECTIVE: SAFETY __________________________________________________ 40 
 
SECONDARY AND EXPLORATORY OBJECTIVES: EVALUATION OF THE IMMUNE RESPONSE ________ 40 
 
ADDITIONAL EXPLORATORY OBJECTIVES __________________________________________ 41 
10 
REMOVAL OF SUBJECTS FROM PROTOCOL THERAPY _____________________________ 42 
 
REMOVAL OF SUBJECTS FROM PROTOCOL THERAPY _________________________________ 42 
 
VOLUNTARY SUBJECT WITHDRAWAL _____________________________________________ 42 
 
PROCEDURE FOR DISCONTINUATION _____________________________________________ 42 
11 
ADVERSE EVENT REPORTING ___________________________________________________ 44 
 
SPONSOR-INVESTIGATOR REPORTING REQUIREMENTS _______________________________ 44 
 
SECONDARY SITE REPORTING REQUIREMENTS _____________________________________ 46 
 
EXCEPTIONS TO EXPEDITED REPORTING __________________________________________ 47 
12 
DATA AND SAFETY MONITORING ________________________________________________ 47 
 
NIH RECOMBINANT DNA ADVISORY COMMITTEE ___________________________________ 48 
13 
STATISTICAL CONSIDERATIONS ________________________________________________ 49 
 
EXPERIMENTAL DESIGN ______________________________________________________ 49 
 
DATA ANALYSIS ____________________________________________________________ 50 
14 
DATA MANAGEMENT __________________________________________________________ 52 
 
ADVERSE EVENT COLLECTION IN THE CASE REPORT FORMS ___________________________ 53 
15 
AUDITING ____________________________________________________________________ 53 
16 
MULTICENTER REGULATORY REQUIREMENTS ____________________________________ 53 
17 
REGULATORY AND ETHICAL OBLIGATIONS _______________________________________ 54 
 
INFORMED CONSENT ________________________________________________________ 54 
 
INSTITUTIONAL REVIEW BOARD _________________________________________________ 54 
 
SUBJECT CONFIDENTIALITY ____________________________________________________ 55 
18 
ADMINISTRATIVE AND LEGAL OBLIGATIONS ______________________________________ 55 
 
STUDY DOCUMENTATION AND RETENTION OF RECORDS _______________________________ 55 
 
POLICY REGARDING RESEARCH-RELATED INJURIES __________________________________ 55 
 
STUDY TERMINATION ________________________________________________________ 56 
19 
REFERENCES _________________________________________________________________ 57 
20 
APPENDICES _________________________________________________________________ 61 
 
ABBREVIATIONS ____________________________________________________________ 61 
 
ECOG/ZUBROD PERFORMANCE STATUS SCALE _____________________________________ 62 
 
NATIONAL CANCER INSTITUTE COMMON TERMINOLOGY CRITERIA FOR ADVERSE EVENTS ______ 62 
20.5 APPENDIX B: Reporting Timelines ________________________________________________ 65 
20.6 APPENDIX C: Washington University Unanticipated Problem Reporting Cover Sheet _____ 69 

Washington University School of Medicine 
Protocol Date 08/03/23, A5 
Siteman Cancer Center 
Pancreatic Cancer SPORE 
Neoantigen DNA vaccines  
 
Page 7 of 69 
 
1 
OBJECTIVES 
This is a phase 1 open-label study to evaluate the safety and immunogenicity of a neoantigen DNA 
vaccine strategy. The neoantigen DNA vaccine strategy is designed to target prioritized neoantigens that 
are present in an individual patient’s pancreatic cancer but not in corresponding normal tissues. The 
vaccines will be formulated as naked plasmid DNA vaccines, and will be administered at a total dose of 4 
mg intramuscularly using an integrated electroporation administration system (TDS-IM system, Ichor 
Medical Systems). The hypothesis of this study is that the neoantigen DNA vaccine strategy will be safe 
for human administration and capable of generating measurable  T cell responses to the neoantigens. 
 
Primary objective 
The primary objective is to assess the safety and feasibility of the neoantigen DNA vaccine approach. 
This trial will be considered feasible if we are able to enroll 15 evaluable patients, successfully perform 
tumor/normal exome sequencing, identify at least 3 neoantigens per patient, and manufacture and 
administer neoantigen vaccines to more than 10 patients. 
 
Secondary objective 
The secondary objective and primary scientific endpoint is to assess the prevalence of antigen-specific T 
cells in the peripheral blood of patients pre- and post-vaccination as measured by ELISPOT.  
 
Exploratory objectives 
Exploratory objectives include measurement of immune response, including the phenotype and functional 
status of neoantigen-specific T cells as measured by CYTOF analysis, as well as assessment of the 
prevalence of antigen-specific T cells in the peripheral blood of patients pre- and post-vaccination as 
measured by flow cytometry. 
An exploratory clinical endpoint is to assess clinical response as measured by disease-free survival and 
overall survival. 
 
2 
BACKGROUND 
 
Pancreatic cancer 
Pancreatic adenocarcinoma is currently the 4th leading cause of cancer related death with the incidence 
of disease expected to increase by 2030 [1, 2]. Despite recent advances in conventional chemotherapy 
regimens for pancreatic cancer there has been little improvement in outcomes for patients facing this 
recalcitrant disease [3, 4]. Currently, surgical resection remains the only curative treatment. However 5-
year survival is achieved in only 20-25% of those with operative disease at time of diagnosis. 
 
Mutational landscape in pancreatic cancer 
Next generation sequencing technologies have been used to characterize the mutational landscape in 
pancreatic cancer. Exome sequencing studies have revealed an average of 63 genetic alterations 
affecting 12 core cellular pathways [5]. The majority of these alterations were point mutations with the 
most frequently mutated genes being KRAS, TP53, and SMAD4. Tumors demonstrated heterogeneity in 
regards to the genomic alterations present, indicating that a spectrum of mutations are present across 
individual patients. 
 
Tumor microenvironment 
Pancreatic cancer has a robust desmoplastic stroma with an abundant leukocyte infiltrate [6, 7]. This 
stroma is implicated in resistance to conventional therapies by limiting effective delivery of therapies to 
malignant cells [8]. The immune infiltrate is predominantly tumor promoting cells derived from the bone 
marrow, including tumor associated macrophages and myeloid derived suppressor cells (MDSC) [9, 10]. 

Washington University School of Medicine 
Protocol Date 08/03/23, A5 
Siteman Cancer Center 
Pancreatic Cancer SPORE 
Neoantigen DNA vaccines  
 
Page 8 of 69 
Furthermore, effector tumor infiltrating lymphocytes are scarce with the majority of tumor infiltrating 
lymphocytes being immunosuppressive FoxP3+ regulatory T-cells (Tregs). 
 
Tumor antigens 
Tumor antigens are often classified as shared tumor antigens and tumor-specific antigens (neoantigens). 
The majority of tumor-specific antigens are now believed to be the result of somatic mutations present in 
the tumor. 
Shared tumor antigens are expressed in multiple cancers, and are often self-differentiation antigens that 
are expressed in a limited subset of normal tissues, but overexpressed in cancers. Examples of shared 
tumor antigens include MAGE (melanoma) [11], prostatic acid phosphatase (prostate cancer) [12], and 
HER2/neu (breast cancer) [13]. Mesothelin is a shared tumor antigen that is highly expressed in 
pancreatic adenocarcinoma [14]. A listeria modified to express mesothelin has been used in pancreatic 
cancer clinical trials and has demonstrated safety and efficacy [15]. Jaffee et al. have characterized the 
immunogenic epitopes for mesothelin for the HLA-A1, HLA-A2, HLA-A3, and HLA-A24 alleles [16, 17]. 
Tumor-specific antigens (or neoantigens) are uniquely expressed in individual cancers, and are typically 
the result of point mutations or other genetic changes that are present only in the tumor (reviewed in [18, 
19]). As such, tumor-specific antigens represent the only antigens that are truly unique to the tumor and 
not expressed in normal tissues. The first human mutant tumor-specific antigen was described in 1995, 
resulting from a point mutation of cyclin-dependent kinase (CDK4) [20]. Since that time additional 
publications have described the expression of neoantigens in melanoma [21], non-small cell lung cancer 
[22] and other human cancers [23]. 
Cancer vaccine strategies targeting neoantigens have substantial theoretical advantages over strategies 
targeting shared tumor antigens. Theoretical advantages include: (1) Targeting neoantigens is potentially 
safer. Neoantigens are expressed only in the tumor, decreasing the risk of autoimmunity. (2) Targeting 
neoantigens is potentially more effective. T cell responses to neoantigens are high in affinity, and are not 
limited by central mechanisms of self-tolerance. (3) Targeting neoantigens potentially limits antigen-loss, 
a common tumor escape mechanism. One of the hallmarks of cancer is genome instability, and one clear 
weakness of cancer vaccines that target a single shared tumor antigen is antigen-loss. Targeting multiple 
neoantigens may preclude antigen loss. In addition, many neoantigens play a functional role in neoplastic 
transformation (driver mutations). (4) Targeting neoantigens is likely to be universally applicable in 
epithelial cancers, including pancreatic cancer. Epithelial cancers appear to have significant numbers of 
nonsynonymous mutations present, suggesting that a personalized vaccine approach could be used in 
most patients with pancreatic cancer, regardless of HLA type.  
We have recently used next generation sequencing technologies to identify and study neoantigens in 
more detail as described in the sections below [24, 25]. 
 
Next generation sequencing 
Robust next-generation sequencing strategies for the identification of neoantigens will be required for the 
successful clinical translation of personalized cancer vaccine strategies. As such, a major focus of our 
research studies has been the development of cost-effective and accurate next-generation sequencing 
strategies to identify neoantigens and validate the expression of these antigens at the mRNA level. 
Initially a cancer genome sequencing approach was used. While cancer whole genome sequencing is 
informative and provides comprehensive information about both the coding and noncoding regions of the 
genome, this level of information may not be necessary for identifying neoantigens, or prioritizing antigens 
for immune intervention. We have now confirmed that tumor/normal exome sequencing is a robust and 
accurate strategy for the identification of neoantigens [24, 25]. Of note, recent studies suggest that 
approximately 40% of mutations identified by cancer exome sequencing are not expressed at the mRNA 
level, so it is important to confirm expression of the mutant allele at the mRNA level. To evaluate mRNA 
expression, we have performed cDNA-capture sequencing analyses. We have confirmed that cDNA-
capture sequencing can be used to successfully confirm expression of sequencing-identified neoantigens 
at the mRNA level. This analysis also provides an estimation of how highly expressed the mutated allele 
is expressed relative to other genes in the tumor. For the phase 1 clinical trial proposed, tumor/normal 
exome sequencing analysis will be used to identify mutations (single nucleotide variants, insertions and 

Washington University School of Medicine 
Protocol Date 08/03/23, A5 
Siteman Cancer Center 
Pancreatic Cancer SPORE 
Neoantigen DNA vaccines  
 
Page 9 of 69 
deletions) present only in the tumor, and cDNA-capture sequencing will be used to confirm mutant allele 
expression and expression level in the tumor mRNA. 
 
Personalized cancer vaccines 
There are two conceptual strategies for creating personalized cancer vaccines targeting neoantigens: a 
candidate epitope strategy, and an unbiased strategy. The candidate epitope strategy uses computer 
algorithms [26, 27] and in vitro studies to predict immunodominant epitopes, which are then integrated 
into a personalized vaccine. In the unbiased strategy, no attempt is made to identify the immunodominant 
epitopes, and all candidate neoantigens are integrated into a personalized vaccine. 
We have considered both the candidate epitope strategy and the unbiased strategy. We believe that the 
candidate epitope strategy is superior to the unbiased strategy for the following reasons. (1) Preliminary 
data from preclinical models and human correlative studies suggest that relatively few sequencing-
identified neoantigens are processed, presented and effectively recognized by the immune system. (2) 
We have now developed and validated algorithms for the prediction and prioritization of sequencing-
identified neoantigens [24, 25]. (3) Targeting a limited number of prioritized sequencing-identified 
neoantigens will facilitate vaccine design and manufacture, and streamline immune monitoring. 
 
Prioritization of sequencing-identified neoantigens 
Of note, we have now developed and validated an epitope prediction algorithm for the prioritization of 
sequencing-identified neoantigens. Once somatic mutations have been identified and mutant mRNA 
expression confirmed/quantified using the sequencing strategies outlined above, neoantigens will be 
prioritized using an epitope prediction algorithm that has been designed to select and prioritize the most 
promising sequencing-identified neoantigens. Currently, the most commonly used CD8 T cell epitope 
prediction algorithm is NetMHC. However, collaborative work conducted by Robert Schreiber, Elaine 
Mardis, Max Artyomov and William Gillanders has shown that a much more accurate prediction comes 
from calculating a median affinity for each sequencing-predicted mutant epitope using multiple available 
epitope prediction algorithms (i.e. NetMHC Pan; ANN; and SMM). We have significantly improved this 
epitope prediction algorithm by applying these filters to the initial prioritized output list: (a) elimination of 
hypothetical proteins; (b) use of an antigen processing algorithm to eliminate epitopes that are not likely 
to be proteolytically produced by constitutive proteasomes or immunoproteasomes; and (c) prioritization 
of “neo-epitopes” identified by a higher affinity binding of the mutant peptide sequence compared to the 
wildtype peptide sequence. The final output of these analyses is a rank-ordered list of the highest to 
lowest priority sequencing-identified neoantigens for each individual patient. In experiments performed 
using preclinical mouse sarcoma models, this refined prediction algorithm has successfully identified the 
major tumor rejection antigens in three out of three tumors tested to date [24, 25]. To our knowledge, this 
is the only algorithm that has been successfully applied to date to cancer vaccine development. Additional 
information about the preclinical validation of the epitope prediction algorithm is provided in Section 5.2 
Nonclinical Studies. MHC class II epitopes can also be accurately predicted using a similar algorithm 
(Artimov, unpublished data). 
Once a rank-ordered list of the highest to lowest priority sequencing-identified mutant tumor-specific 
peptide antigens is generated, we will generate tetramers corresponding to the top 5-10 class I 
neoantigens using a photo-activated peptide tetramer protocol. We will stain T cells from the peripheral 
blood of subjects using a state-of-the-art CyTOF processor. Mutant epitopes associated with positive 
tetramer staining will be prioritized for the generation of neoantigen DNA vaccines. Once the candidate 
mutant epitopes have been verified, multivalent DNA vaccine construction will commence. 
 
Polyepitope DNA vaccines 
The neoantigen DNA vaccine strategy is based on the DNA vaccine platform. The observation that direct 
administration of recombinant DNA can generate potent immune responses established the field of DNA 
vaccines in the early 1990s [28-33]. Since that time, DNA vaccines have remained an area of intense 
research interest, and vaccines targeting infectious disease agents and cancers have progressed into 
clinical trials. Advantages of the DNA vaccine platform include the remarkable safety profile of DNA 
vaccines, and the relative ease of manufacture relative to proteins and other biologics. Perhaps most 
important, however, is the molecular flexibility of the DNA vaccine platform, with the ability to genetically 

Washington University School of Medicine 
Protocol Date 08/03/23, A5 
Siteman Cancer Center 
Pancreatic Cancer SPORE 
Neoantigen DNA vaccines  
 
Page 10 of 69 
manipulate encoded antigens, and/or incorporate other genes to amplify the immune response [34, 35]. 
The molecular flexibility of the DNA vaccine platform allows us to target multiple mutant tumor-specific 
antigens using a single polyepitope DNA vaccine. Polyepitope DNA vaccines integrate multiple epitopes 
in a single construct (Figure 1). We have optimized the polyepitope DNA vaccine platform to maximize 
antigen presentation of neoantigens by integrating a mutant ubiquitin molecule. Because MHC class I 
binding peptides are initially processed in the cytosol by the ubiquitin/proteasome pathway, we have 
integrated a mutant form of ubiquitin (UbG76V). Fusion of UbG76V to the N-terminus of the polyepitope 
construct promotes epitope generation and display. 
 
 
 
 
Electroporation 
Recent research provides valuable insights into why preclinical studies of DNA vaccines have been 
successful in rodents, but less successful in larger animals. One difficulty is scaling up DNA vaccine dose 
and injection volume [36]. In rodents, hydrostatic pressure from injecting DNA in a relatively large liquid 
volume significantly improves cellular uptake of DNA and antigen expression [37]. This effect is reduced 
in larger animals as the relative volume injected and hydrostatic pressure is reduced. Electroporation 
dramatically increases DNA uptake by muscle cells, antigen expression, and immunogenicity [38-41]. Of 
particular note, electroporation has now been used successfully in non-human primates, with responses 
at levels previously not observed with other DNA vaccine approaches and similar to or superior to 
responses induced by live vectors [42-49]. The importance of electroporation to the successful clinical 
translation of DNA vaccines was recently highlighted and emphasized in a high-profile review in Nature 
Reviews [50], and in the introduction to a special issue of Vaccine [51]. We have established a 
collaboration with Ichor Medical Systems and will use the TDS-IM electroporation device in the proposed 
phase 1 clinical trial. 
 Clinical trial design and dose considerations 
The early clinical development paradigm for chemotherapeutic agents has significantly influenced the 
development of therapeutic cancer vaccines. However, there are major differences between these two 
classes of therapeutics that have important implications for early clinical development. Specifically, the 
phase 1 concept of dose escalation to find a maximum-tolerated dose does not apply to most therapeutic 
cancer vaccines. Most therapeutic cancer vaccines are associated with minimal toxicity at a range that is 
feasible to manufacture or administer, and there is little reason to believe that the maximum-tolerated 
dose is the most effective dose. 
Figure 1: Polyepitope DNA vaccine design. Neoantigen DNA vaccines integrate multiple epitopes 
in a single construct. We have optimized the polyepitope DNA vaccine platform to maximize antigen 
presentation of the sequencing-identified neoantigens by integrating a mutant ubiquitin molecule. 

Washington University School of Medicine 
Protocol Date 08/03/23, A5 
Siteman Cancer Center 
Pancreatic Cancer SPORE 
Neoantigen DNA vaccines  
 
Page 11 of 69 
In a recent article from the biostatistics literature, Simon et al. write that "the initial clinical trial of many 
new vaccines will not be a toxicity or dose-ranging trial but rather will involve administration of a fixed 
dose of vaccine … in most cases the dose selected will be based on preclinical findings or practical 
considerations. Using several dose levels in the initial study to find the minimal active dose or to 
characterize the dose-activity relationship is generally not realistic" [52]. 
To date, the Ichor TriGrid™ Delivery System has been used as the means for delivery in nine DNA 
vaccine candidates. In the safety and toxicology studies, notable findings associated with electroporation-
mediated delivery of DNA vaccines were limited to localized inflammatory responses of mild to moderate 
severity at the site of administration. Please see Sections 5.12.3 and 5.12.4 for a detailed description of 
the TDS-IM clinical experience to date. 
We propose a vaccine dose of 4 mg based primarily on practical considerations and previous experience 
with the TDS-IM device. This dose is the within the range of DNA doses previously tested in the clinical 
setting with the TDS-IM device (0.2-8.0 mg) and, given the concentration of the neoantigen DNA vaccine 
(2.0 mg/mL), this dose is the maximal dose that can be administered feasibly by two administrations of 
electroporation. 
 Challenges of sequencing pancreatic cancers 
The challenges of performing massively parallel sequencing (MPS) assays from clinical pancreatic cancer 
samples are considerable. At its essence, pancreatic adenocarcinoma has a low cellularity, with many 
stromal cells interspersed with the pancreatic cancer cells. A pathologist who is a specialist in 
gastrointestinal cancers will survey the tumor to identify islands of high tumor cellularity and take 1mm 
punches from formalin fixed paraffin embedded (FFPE) tissues.  By selecting these areas for sequencing 
we are able to enrich the tumor purity to the level that allows neoantigen identification. Additionally, since 
the tissues have been previously subjected to FFPE, some degradation of the nucleic acids may result, 
further complicating the sample quality toward MPS. 
Nevertheless, our group has worked extensively to optimize MPS library construction techniques that 
enable high diversity libraries to be constructed from as little as 10 ng of input DNA. In particular, we 
currently utilize the Swift Biosciences Accel-NGS 2S kit that is optimized for low input DNA from FFPE 
samples with an input range of 10-50 ng of isolated DNA. The resulting libraries have proven suitable for 
subsequent exome capture by hybridization such as the Integrated DNA Technologies (IDT) Rapid 
Exome reagent. This commercially available reagent has superior performance on FFPE-derived DNA 
libraries, and has a rapid hybridization time of only 4 hours, permitting a one-day turn around for library 
construction, quantitation, and hybrid capture prior to sequencing. 
Another challenge of pancreatic adenocarcinoma is the rapid degradation of RNA that often occurs during 
surgery. Our surgical team has been evaluating whether the RNA stability can be influenced by changes 
to the surgical procedure that reduce ischemia time during surgery. In our early experience with several 
samples, this alteration in the surgical procedure leads to dramatic improvements to the RNA stability as 
gauged by RNA integrity (RIN) values obtained from our post-isolation QC procedures. In turn, the MPS 
libraries generated from these pancreatic RNAs are of high quality and diversity. 
 
3 
PATIENT SELECTION 
 
Inclusion criteria 
A patient will be eligible for the trial only if ALL of the following criteria apply: 
1. Histologically or cytologically confirmed diagnosis of pancreatic adenocarcinoma; mixed 
histology will be included as long as the predominant histology is adenocarcinoma. 
2. Completed an R0 or R1 surgical resection as determined by pathology  
3. Pathology review demonstrates tumor cellularity no less than 30% in quantities sufficient to 
obtain 6-8 1mm biopsies from the original FFPE blocks.  
4. At least 18 years of age. 

Washington University School of Medicine 
Protocol Date 08/03/23, A5 
Siteman Cancer Center 
Pancreatic Cancer SPORE 
Neoantigen DNA vaccines  
 
Page 12 of 69 
5. Life expectancy of > 12 months. 
6. ECOG performance status ≤ 2 
7. Normal bone marrow and organ function as defined below: 
a. WBC 
≥3,000/μL 
b. absolute neutrophil count 
≥1,500/μL 
c. platelets 
≥100,000/μL 
d. total bilirubin 
≤2.5 X institutional upper limit of normal 
e. AST/ALT 
≤2.5 X institutional upper limit of normal 
f. 
creatinine 
≤1.5 X institutional upper limit of normal 
8. International Normalized Ratio (INR) and activated partial thromboplastin time (PTT) < 1.5 x 
ULN provided the patient is not on anticoagulation therapy. 
9. Patients who have had a stent placed for biliary obstruction can be included in the study 
provided serum bilirubin at time of enrollment is within protocol limits. 
10. Women of child-bearing potential and men must agree to use adequate contraception (hormonal 
or barrier method of birth control, abstinence) prior to study entry and for the duration of study 
participation. Should a woman become pregnant or suspect she is pregnant while participating 
in this study, she must inform her treating physician immediately. 
11. Able to understand and willing to sign an IRB approved written informed consent document. 
Patients may be consented prior to receiving adjuvant therapy, or during the course of adjuvant therapy.  
Adjuvant therapy must meet the following criteria below for enrollment to the trial: 
a. Initiation of adjuvant chemotherapy within 12 weeks of surgery 
b. Completion of at least 4 months of adjuvant chemotherapy with 
gemcitabine/capecitabine or similar adjuvant chemotherapy at the discretion of the 
patient’s medical oncologist.  
c. Additional chemoradiation therapy as recommended by the patient’s medical oncologist. 
d. Reimaging within 4 weeks of last dose of chemotherapy demonstrates no evidence of 
recurrent disease and CA 19-9 is less than 92.5 u/mL. 
e. Dose modifications and/or delays in adjuvant chemotherapy is at the discretion of the 
treating physician 
2. There is a 1 week washout prior to Day 1 of vaccine for patients on daily systemic steroids at 
doses exceeding 10 mg prednisone. 
 
Exclusion criteria 
A patient will be ineligible for inclusion in this study if ANY of the following criteria apply: 
1. Evidence of neuroendocrine tumor, duodenal adenocarcinoma, or ampullary adenocarcinoma.  
2. Received neoadjuvant chemotherapy for their pancreatic adenocarcinoma. 
3. Evidence of disease recurrence or metastasis following surgical resection at any time prior to the 
first vaccination administration. Most patients will undergo restaging midway through adjuvant 
chemotherapy and at the completion of therapy; however, timing of imaging is at the discretion of 
the patient’s medical oncologist. 
4. History of other malignancy ≤ 3 years previous with the exception of basal cell or squamous cell 
carcinoma of the skin which were treated with local resection only or carcinoma in situ of the 
cervix. 
5. Receiving any other investigational agents, or has received an investigational agent within the 
last 30 days. 
6. Known allergy, or history of serious adverse reaction to vaccines such as anaphylaxis, hives, or 
respiratory difficulty. 

Washington University School of Medicine 
Protocol Date 08/03/23, A5 
Siteman Cancer Center 
Pancreatic Cancer SPORE 
Neoantigen DNA vaccines  
 
Page 13 of 69 
7. Acute or chronic, clinically significant hematologic, pulmonary, cardiovascular, hepatic renal, 
and/or other functional abnormality that would jeopardize the health and safety of the participant 
as determined by the investigator based on medical history, physical examination, laboratory 
values, and/or diagnostic studies. 
8. A psychiatric illness/social situations that would limit compliance with study requirements as 
determined by the investigator from the medical history, physical exam, and/or medical record 
9. History of syncopal or vasovagal episode as determined by medical record and history in the 12 
month period prior to first vaccination administration. 
10. Individuals in whom a skinfold measurement of the cutaneous and subcutaneous tissue for 
eligible injection sites (left and right medial deltoid region) exceeds 40 mm. 
11. Individuals in whom the ability to observe possible local reactions at the eligible injection sites 
(deltoid region) is, in the opinion of the investigator, unacceptably obscured due to a physical 
condition or permanent body art. 
12. Therapeutic or traumatic metal implant in the skin or muscle of either deltoid region. 
13. Any chronic or active neurologic disorder, including seizures and epilepsy, excluding a single 
febrile seizure as a child. 
14. Current use of any electronic stimulation device, such as cardiac demand pacemakers, automatic 
implantable cardiac defibrillator, nerve stimulators, or deep brain stimulators. 
15. Prior or currently active autoimmune disease requiring management with immunosuppression. 
This includes inflammatory bowel disease, ulcerative colitis, Crohn’s disease, systemic vasculitis, 
scleroderma, psoriasis, multiple sclerosis, hemolytic anemia, immune-mediated 
thrombocytopenia, rheumatoid arthritis, systemic lupus erythematosus, Sjögren’s syndrome, 
sarcoidosis, or other rheumatologic disease or any other medical condition or use of medication 
(e.g., corticosteroids) which might make it difficult for the patient to complete the full course of 
treatments or to generate an immune response to vaccines. In the case of asthma or chronic 
obstructive pulmonary disease taking inhaled corticosteroids that does not require daily systemic 
corticosteroids is acceptable. Additionally, local acting steroids (topical, inhaled, or intraarticular) 
will be allowed. Patients on intermittent or short course steroids will be allow if the dose does not 
exceed 4 mg of dexamethasone (or equivalent) per day for > 7 consecutive days. Any patients 
receiving steroids should be discussed with the PI to determine if eligible. 
16. Pregnant and/or breastfeeding. 
17. Known HIV-positive status. These patients are ineligible because of the potential inability to 
generate an immune response to vaccines. 
 
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 
 
Prior to registration 
4.1.1 
Subject recruitment 
The methods used for recruitment of subjects in the study will be devoid of any procedures that may be 
construed as coercive. The recruitment process will not involve any restrictions based on social or 
demographic factors including age, or ethnic characteristics of the subject population. However, the 
composition of the study subject population will depend on patient sources available to the investigators. 
Subjects will be identified and recruited for this study as follows: 
Patients will be recruited from Washington University’s Siteman Cancer Center and Johns Hopkins 
Sidney Kimmel Comprehensive Cancer Center outpatients or patient referrals by our community 

Washington University School of Medicine 
Protocol Date 08/03/23, A5 
Siteman Cancer Center 
Pancreatic Cancer SPORE 
Neoantigen DNA vaccines  
 
Page 14 of 69 
oncologists to the principal investigator and co-investigators. Patients must be willing and able to give 
their written informed consent indicating that they are aware of the investigational nature of the study. 
After a patient is deemed eligible for study, the principal investigator (or co-investigators), or another 
delegated individual, will discuss the Washington University/Johns Hopkins IRB-approved informed 
consent with the patient. This written informed consent will be signed and dated by the patient and the 
principal investigator (or co-investigators), or another delegated individual. The original consent will be 
placed in the patient’s permanent record and a copy will be given to the patient. 
Washington University School of Medicine (WUSM)/Johns Hopkins Medicine (JHM) has an approved 
Multiple Project Assurance of Compliance with Department of Health and Human Services Regulations 
for the Protection of Human Research Subjects on file with the Office for Human Research Protection 
(OHRP). The Human Research Protection Office Policies and Procedures for Protection of Human 
Research Subjects details all policies and procedures for the protection of human research subjects and 
can be obtained upon request from the Human Research Protection Office. 
4.1.2 
Compliance and understanding 
All patients who present with a diagnosis of pancreatic cancer will be screened for eligibility for entry into 
the study at their postoperative follow up with their surgeon or initial postoperative appointment with their 
medical oncologist, or during their adjuvant treatment with their medical oncologist. As in all trials, the 
goal is to achieve a high level of compliance with protocol requirements by assuring, during the eligibility 
assessment, that the potential subject is fully informed and agrees to the protocol requirements. In 
addition, subjects with a strong likelihood of non-adherence such as difficulties in adhering to follow-up 
schedule due to geographic distance from the Siteman Cancer Center/Sidney Kimmel Comprehensive 
Cancer Center, should not knowingly be registered. Adherence of the Siteman Cancer Center/Sidney 
Kimmel Comprehensive Cancer Center staff to careful assessment of the subject’s understanding of the 
trial and a clinical center environment which supports the continued commitment of the subjects are 
essential for the trial to be successfully completed. 
4.1.3 
Presentation of informed consent 
Consent will be obtained by either the principal investigator or by individuals approved by the principal 
investigator and whose names and copy of their curriculum vitae have been filed. The initial consent 
should be the IRB-approved version corresponding to the version of the protocol approved when the 
screening was initiated. Informed consent is to be obtained from the subject according to Section 15.1 
Informed Consent of this protocol. 
 
Registration procedures 
Patients must not start any protocol intervention prior to registration through the Siteman Cancer 
Center. 
Registration for this trial will be a two-step process.  Step 0 will be screening and confirmation of eligibility 
for vaccine creation.  Step 1 will be screening and confirmation of eligibility for vaccine administration.  
See Section 3:  Patient Selection for eligibility criteria for each step and Section 8:  Study Calendar for 
screening and procedures for each step. 
Step 0 screening must occur within 16 weeks of surgical resection, and Step 0 enrollment can occur up to 
four weeks after initiation of adjuvant chemotherapy.  Step 1 screening will occur when the patient’s 
vaccine is ready or nearly ready, but must be within 14 days of the first vaccine administration. 
The following steps must be taken before registering patients to this study: 
(1) 
Confirmation of patient eligibility by Washington University (applies to both Step 0 and Step 1) 
(2) 
Registration of patient in the OnCore Cancer Center database (This will be done by Washington 
University research coordinator.  Step 0 enrollment will be “On Study” in the OnCore database, 
and Step 1 enrollment will be “On Treatment” in the OnCore database.) 
(3) 
Assignment of unique patient number (UPN) (assigned as Step 0 only) 

Washington University School of Medicine 
Protocol Date 08/03/23, A5 
Siteman Cancer Center 
Pancreatic Cancer SPORE 
Neoantigen DNA vaccines  
 
Page 15 of 69 
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. 
 
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 two business days prior to registering patient (if a quicker turnaround is 
needed, contact primary study coordinator at Siteman Cancer Center): 
(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 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 first injection 
(8) 
Completed eligibility checklist, signed and dated by a member of the study team 
(9) 
Copy of appropriate redacted source documentation confirming patient eligibility 
 
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. 
 
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. 
 
5 
INVESTIGATIONAL AGENT 
 
Chemical name and structure 
Neoantigen DNA vaccines will be designed in the Gillanders laboratory and manufactured at the Siteman 
Cancer Center Biological Therapy Core Facility based on the following general steps:  
(1) Pancreatic adenocarcinoma tissue and normal lymphocytes will be obtained from pancreatic cancer 
patients who are eligible for the phase 1 clinical trial.  
(2) A pancreas specific pathologist will review the operative specimen slides to verify tumor cellularity 
and quantity sufficient for proceeding 
(3) The corresponding FFPE blocks will be punched with a disposable 1mm biopsy punch. 6-8 full 
thickness punches will be taken from areas of high tumor cellularity and divided evenly into two DNA 
LoBind Eppendorf tubes (one tube for DNA and one for RNA).  

Washington University School of Medicine 
Protocol Date 08/03/23, A5 
Siteman Cancer Center 
Pancreatic Cancer SPORE 
Neoantigen DNA vaccines  
 
Page 16 of 69 
(4) DNA and RNA are subsequently extracted in the Washington University Center for Human 
Immunology and immunotherapy Programs (CHIIPS) core.  
(5)  Tumor/normal exome sequencing and tumor cDNA-capture sequencing, if feasible, will be performed 
to identify candidate neoantigens. Candidate neoantigens will be prioritized based on epitope 
prediction algorithms as outlined previously.  
(6) Mesothelin expression will be confirmed by RNA analysis, if patients are confirmed to express 
mesothelin and are HLA-A1, HLA-A2, HLA-A3, or HLA-24, the corresponding mesothelin epitopes will 
be added to the vaccine.  
(7) Personalized polyepitope inserts integrating the prioritized neoantigens and mesothelin epitopes will 
be designed in the Gillanders laboratory and then synthesized and cloned into the pING parent vector 
by Blue Heron Biotech. 
(8) Master cell banks will be established and validated at WUSM. 
(9) The neoantigen DNA vaccines will be manufactured and vialed at WUSM. 
(10) The neoantigen DNA vaccines will undergo product release tests at WUSM and other designated 
sites (See Table 3) prior to investigational use. 
The neoantigen DNA vaccines will be manufactured in the SCC Biologic Therapy Core facility. Standard 
Operating Procedures for the GMP manufacture of the personalized vaccines have been established and 
are in accordance with “CGMP for Phase 1 Investigational Drugs 2008,” and “Considerations for Plasmid 
DNA Vaccines for Infectious Disease Indications 2007.” 
Each polyepitope DNA vaccine drug product is composed of a deoxyribonucleic acid (DNA) plasmid 
purified from E. Coli. The pING parent vector was obtained from Alan Houghton, M.D. at Memorial Sloan 
Kettering Cancer Center (Figure 2). The pING vector has been used extensively in preclinical DNA 
vaccine studies [53, 54], and in human clinical trials, including a clinical trial at WUSM (ClinicalTrials.gov 
identifier: NCT00807781). This trial using a mammaglobin A DNA breast cancer vaccine is based on the 
pING vector as well and was manufactured at WUSM in the SCC Biologic Therapy Core Facility using 
manufacturing processes almost identical to the ones outlined here. 
The pING vector contains the following elements: (1) a eukaryotic promoter and enhancer from the 
Towne strain of CMV; (2) a polylinker region to facilitate cloning of a variety of DNA fragments; (3) donor 
and acceptor splice sites and a poly adenylation signal sequence derived from the bovine growth 
hormone gene; (4) the ColE1 origin of replication and (5) a gene conferring kanamycin resistance. With 
the exception of the kanamycin resistance gene, which was cloned as a PstI fragment from the plasmid 
pUC4, all other gene segments were amplified by polymerase chain reaction (PCR). 
The role of each element is as follows: (1) the CMV promoter/enhancer enables high-level expression of 
polyepitope insert in mammalian cells; (2) the polylinker region serves as a multiple cloning site for easy 
insertion of genes, in this case the polyepitope insert; (3) the polyadenylation signal sequence facilitates 
efficient transcription termination and polyadenylation of mRNA; (4) the origin of replication allows for 
high-copy number replication and growth in E. coli; (5) the kanamycin resistance gene provides for 
selection in E. coli; and (6) the prokaryotic T7 promoter of the pING plasmid produces high levels of 
polyepitope DNA transcripts in E. coli in the presence of bacterial T7 RNA polymerase. Of note, the 
kanamycin resistance gene is cloned in the opposite orientation from the polyepitope insert to limit 
transcription of kanamycin from the CMV promoter in human cells. 
 

Washington University School of Medicine 
Protocol Date 08/03/23, A5 
Siteman Cancer Center 
Pancreatic Cancer SPORE 
Neoantigen DNA vaccines  
 
Page 17 of 69 
 
 
The personalized inserts will be designed in the Gillanders Laboratory based on the results of next-
generation sequencing and epitope prediction algorithms from each individual patient. Personalized 
polyepitope inserts will incorporate up to 20 prioritized neoantigens. The personalized polyepitope inserts 
will be synthesized by Blue Heron Technologies and cloned into the pING parent vector as detailed 
below. 
 
Nonclinical studies 
5.2.1 
Overview 
We propose a phase 1 clinical trial of a neoantigen DNA vaccine strategy. The neoantigen DNA vaccine 
strategy is designed to target neoantigens present in the pancreatic cancer, but absent in normal tissues. 
One of the reasons that we have pursued clinical development of a neoantigen DNA vaccine strategy 
targeting neoantigens is because we believe that this strategy has the potential to be safer than strategies 
targeting shared tumor antigens. Shared tumor antigens are typically expressed at high levels in the 
tumor, but are also typically expressed at lower levels in some normal tissues. Expression of shared 
tumor antigens in normal tissues may increase the risk of autoimmunity. Neoantigens are present only in 
the tumor. In addition, our next-generation sequencing-based epitope prediction algorithm prioritizes 
epitopes where the mutant epitope (but not the wildtype epitope) can bind to restricting HLA molecules. 
This decreases the potential that immune responses targeting neoantigens will be cross-reactive with 
wildtype antigens. 
We do not think that GLP safety and toxicology studies will provide significant insight into the safety of the 
neoantigen DNA vaccine strategy. First, it is impossible to know a priori what mutations will be present 
and/or prioritized in individual patients. We estimate that there are as many as 7 million potential 
neoantigens that could be targeted by our approach. Only a limited number of mutations could be 
targeted in GLP safety and toxicology studies. Second, to our knowledge, no mammary tumor models 
exist that would be relevant for GLP safety and toxicology studies. Third, the pING parental vector has 
proven to be safe in phase 1 clinical trials. 
We are not proposing to perform GLP biodistribution and integration studies at this stage in development. 
There is extensive GLP information available about the biodistribution and integration of DNA vaccines 
Figure 2: Polyepitope DNA vaccine design. Polyepitope DNA vaccines are based on the pING 
parent vector. The pING vector contains the following elements: (1) a eukaryotic promoter and 
enhancer from the Towne strain of CMV; (2) a polylinker region to facilitate cloning of a variety of DNA 
fragments; (3) donor and acceptor splice sites and a poly adenylation signal sequence derived from 
the bovine growth hormone gene; (4) the ColE1 origin of replication; (5) a gene conferring kanamycin 
resistance, and (6) a polyepitope insert integrating the prioritized neoantigens. 

Washington University School of Medicine 
Protocol Date 08/03/23, A5 
Siteman Cancer Center 
Pancreatic Cancer SPORE 
Neoantigen DNA vaccines  
 
Page 18 of 69 
following electroporation with the TriGrid device. This includes GLP information about biodistribution and 
integration of DNA vaccines using the pING parent vector. Additional information about the biodistribution 
and integration of DNA vaccines following electroporation with the TriGrid device is summarized in the 
IND application. 
5.2.2 
Nonclinical studies 
It has long been known that there is a dynamic relationship between the immune system and cancer. This 
dynamic relationship has been studied in detail, ultimately resulting in the establishment of the cancer 
immunoediting concept [55-62].  
We have recently focused on defining the antigens recognized by the immune system during the cancer 
immunoediting process. These studies, summarized below, demonstrate that neoantigens are important 
tumor rejection antigens, and provide strong support for our personalized pancreatic cancer vaccine 
strategy. Specifically, we have developed next-generation sequencing and epitope prediction algorithms 
to identify and prioritize neoantigens. We will use these algorithms in the proposed clinical trial. The 
preclinical data supporting the use of these algorithms are presented below. 
In initial studies we used a combination of next-generation sequencing and epitope prediction algorithms 
to identify neoantigens in the d42m1 MCA sarcoma line. These algorithms identified one particular 
mutation (an R913L mutation of SPTBN2) as a top candidate, and subsequent analyses confirmed that 
this mutant tumor-specific antigen functioned as an immunodominant tumor rejection antigen. These 
studies were published in Nature [24]. 
The d42m1 MCA sarcoma is an unedited tumor, and would therefore be expected to express strong 
tumor rejection antigens. We have since turned our attention to examining the epitope landscape in 
edited MCA sarcomas that develop in immunocompetent wildtype mice. Specifically, we have asked the 
following questions: (1) Can the next-generation sequencing and epitope prediction algorithms be used 
more broadly to identify and prioritize important neoantigens in less immunogenic tumors? (2) Can the 
next-generation sequencing and epitope prediction algorithms be used to prioritize antigens for immune 
targeting and/or personalized vaccine therapy? 
To address these questions we focused initial efforts on d42m1-T3. d42m1-T3 is a clone of d42m1 that 
lacks the immunodominant rejection antigen, mutant SPTBN2, and forms progressively growing tumors in 
wildtype mice. We specifically chose the d42m1-T3 clone because d42m1-T3 shares with naturally edited 
sarcomas the ability to form progressively growing tumors in wildtype mice and shows a similar sensitivity 
to checkpoint blockade.  
To identify and prioritize mutant tumor specific antigens from the d42m1-T3 we used optimized next-
generation sequencing and epitope prediction algorithms. Specifically, we pipelined the candidate mutant 
tumor-specific antigen sequences into four different MHC class I epitope prediction algorithms and 
calculated the median predicted affinity for binding to the relevant class I MHC alleles. We then applied 
filters that account for proteasomal processing of the antigen and differences in MHC class I binding 
affinity between mutant and native sequences to prioritize the neoantigens. We also deprioritized 
hypothetical Riken proteins.  
Of the top 61 prioritized candidates, 20 were eliminated by the filtering process; including two of the top 
four candidates. Of those that remained, two [G1254V Laminin subunit α4 (mLama4) and A506T alpha-
1,3 glucosyltransferase (mAlg8)] were clearly favored above the others based on predicted binding 
affinity.  
To test whether these two “best” neoantigens were biologically relevant, we generated tumor-specific 
CD8 T cell lines from the spleens of three independent mice that had rejected d42m1-T3 cells after anti-
PD-1 therapy and showed that each T cell line (CTL-62, CTL-73, CTL-74) displayed specificity for d42m1-
T3 but not an unrelated sarcoma, F244. To determine if the “prioritized” neoantigens were recognized by 
anti-d42m1-T3 T cell lines, we incubated 8 amino acid synthetic peptides corresponding to each of the 
top 61 initially predicted H-2Kb neoantigens with irradiated splenocytes and CTL-74 T cells and monitored 
IFNγ production. The mLama4 and mAlg8 peptides strongly stimulated CTL-74 T cells, with mLama4 
inducing ~10x more IFNγ than mAlg8. No other predicted mutant epitope induced significant levels of 
IFNγ production in this assay. Similar results were obtained with the other two d42m1-T3 specific CD8 T 

Washington University School of Medicine 
Protocol Date 08/03/23, A5 
Siteman Cancer Center 
Pancreatic Cancer SPORE 
Neoantigen DNA vaccines  
 
Page 19 of 69 
cell lines. Subsequent dose response experiments showed that mLama4 stimulated the tumor-specific T 
cell lines to a greater extent than mAlg8 and that the T cells reacted specifically with mutant but not native 
peptides.  
We then used four experimental systems to confirm that our optimized epitope prediction algorithms 
accurately prioritized neoantigens. First, together with the groups of Hans-Georg Rammensee in 
Tübingen and Ruedi Abersold in Zurich we detected mLama4 and mAlg8 peptides bound to H-2Kb on 
d42m1-T3 tumor cells. To our knowledge this is the first time that mutant class I epitopes have been 
detected bound to tumor cell-associated MHC class I. Second, using PE-labeled H-2Kb tetramers carrying 
mLama4 or mAlg8 peptides, CD8 T cells with specificities for these two epitopes were found to 
accumulate in d42m1-T3 tumors in αPD-1 treated mice and reached peak values just prior to tumor 
rejection on day 12. Consistent with the results of the T cell stimulation experiments, mLama4-specific T 
cells were present in significantly higher numbers in the tumor than mAlg8-specific T cells. No mLama4- 
or mAlg8-specific T cells were observed in irrelevant, checkpoint blockade-sensitive F244 tumors. Third, 
vaccination of naïve WT mice with mutant-Lama4 or mutant-Alg8 short peptide vaccines (8mer) induced 
strong CD8 T cell responses that were specific for the mutant, but not the WT epitope (mLama4 = 1650 
SFC/106 cells vs. wtLama4 = 75 SFC/106 cells; mAlg8 =606 SFC/106 cells vs. wtAlg8 = 50 SFC/106 cells). 
Fourth, prophylactic vaccination of mice with long peptides (~30mer) corresponding to either the mLama4 
epitope alone, or both the mLama4 and mAlg8 epitopes induced protection against subsequent challenge 
with d42m1-T3 tumor cells. The combined peptide vaccine was more protective than the vaccine 
containing the mLama4 long peptide alone (Figure 5). 
Of note, immune responses to both mLama4 and mAlg8 were also observed after vaccination with a 
polyepitope DNA vaccine encoding mLama4, mAlg8, and several additional epitopes. Mice were 
vaccinated three times and the immune response to mLama4, mAlg8 and control peptides was assessed 
by ELISPOT using splenocytes from vaccinated mice. Vaccination with polyepitope DNA vaccines 
induced responses that were similar in magnitude as synthetic long peptide vaccines encoding mLama4 
or mAlg8 (data not shown). 
5.2.3 
Rationale for no GLP safety studies 
We do not think that GLP safety and toxicology studies will provide significant insight into the safety of the 
neoantigen DNA vaccine strategy. 
First, it is impossible to know a priori what mutations will be present and/or prioritized in patients. Next-
generation sequencing of epithelial cancers has demonstrated that there are very few recurrent mutations 
present. Mutations can be present in any one of the approximately 20,000 protein-coding genes present 
in the human genome. Even more problematic is that the mutations targeted could be anywhere in the 
corresponding protein. It has been estimated that the average length of a protein in humans is 362 AA. 
Thus, there are potentially 7,240,000 potential neoantigens that could be targeted by our approach. This 
includes only point mutations and does not include mutations resulting from indels. If studies are 
performed in a preclinical model, only a limited number of mutations will be targeted. For example if we 
design a neoantigen DNA vaccine specific for a murine epithelial cancer targeting 5 genes, this would 
represent only 5 of 7,240,000 potential neoantigens. Even if 1000 GLP safety and toxicology studies were 
performed, each targeting 5 neoantigens, it would still provide information on < 0.07% of potential 
neoantigens. 
Second, to our knowledge, no pancreatic cancer models exist that would be relevant for the proposed 
preclinical studies. In order to study neoantigens, sequencing analyses of paired tumor/normal tissues are 
required. As such, we would need to study spontaneous tumors in mice, as tumors propagated as cell 
lines do not have corresponding normal DNA to evaluate. There are very few models of spontaneous 
pancreatic cancer development in wildtype mice. Spontaneous tumors do develop in genetically 
engineered mice, but these oncogene-driven tumors in genetically engineered murine models of cancer 
typically have a very limited number of mutations. We have performed extensive studies in genetically 
engineered mice, and have found that there are very few mutations present in tumors derived from these 
mice. For example, we have performed extensive studies in the p53-null transplant mammary tumor 
model. p53 is a tumor suppressor gene and plays an important role in maintaining genome stability. As 
such one might expect that there would be a significant number of mutations present in p53-null 
transplant mammary tumors. However, we have sequenced > 10 p53-null transplant mammary tumors 

Washington University School of Medicine 
Protocol Date 08/03/23, A5 
Siteman Cancer Center 
Pancreatic Cancer SPORE 
Neoantigen DNA vaccines  
 
Page 20 of 69 
and have found that there are only a limited number of mutations present in each tumor. This is not at all 
representative of human pancreatic cancers where significantly more mutations are present. Of note, a 
significant number of mutations must be present to reliably identify neoantigens that are immunogenic, as 
many candidate neoantigens are not processed and presented by the immune system, or cannot be 
recognized efficiently by T cells. If only a limited number of mutations is present, it is possible that no 
neoantigens will be identified that are immunogenic. For meaningful GLP safety and toxicology studies of 
a neoantigen DNA cancer vaccine, one of the key considerations is to assess any potential toxicity 
associated with immune responses to the vaccine. If no neoantigens are identified that are immunogenic, 
the GLP safety and toxicology studies will have only limited value as no immune response to the mutant 
tumor-specific antigen will be generated. As such, we are not aware of any pancreatic cancer tumor 
models in mice that would be appropriate for the GLP safety and toxicology studies. 
5.2.4 
Rationale for no GLP biodistribution studies 
We are not proposing to perform GLP biodistribution and integration studies at this stage in development. 
There is extensive GLP information available about the biodistribution and integration of DNA vaccines 
following electroporation with the TriGrid device. This includes GLP information about biodistribution and 
integration of DNA vaccines using the pING parent vector. The information available about the 
biodistribution and integration of DNA vaccines following electroporation with the TriGrid device is 
detailed below. 
 
Sequencing pipeline 
Robust next-generation sequencing strategies for the identification of neoantigens will be required for the 
successful clinical translation of personalized cancer vaccine strategies. As such, a major focus of our 
research studies has been the development of cost-effective and accurate next-generation sequencing 
strategies to identify neoantigens and validate the expression of these antigens at the mRNA level.  
The first step in the sequencing pipeline is exome sequencing of pancreatic cancer and normal DNA. 
Exome fragments are captured using Nimblegen's "VCRome" exome capture reagent. Background DNA 
is washed away while the bound exome DNA is eluted and sequenced. Separate libraries are made from 
the pancreatic cancer and normal DNA and processed independently. Exome sequencing is performed 
using the Illumina platform. 
Exome sequences from pancreatic cancer and normal DNA are compared separately to the human 
reference sequence and then to one another to identify somatic variation. VarScan 2 software is used to 
detect misaligned sequences and identify structural variants in the pancreatic cancer DNA.  
The second step in the sequencing pipeline is cDNA-capture sequencing. To validate the results of the 
exome sequencing, and confirm expression of the somatic mutations in the pancreatic cancer, cDNA-
capture sequencing of the pancreatic cancer RNA will be performed. cDNA-capture sequencing is very 
similar to RNA sequencing, but cDNA is captured prior to sequencing to enrich for mRNA. cDNA-capture 
sequencing is a sensitive and accurate methodology to detect expression of somatic mutations at the 
mRNA level in pancreatic cancer. cDNA-capture sequencing is performed using the Illumina platform.  
The third step in the sequencing pipeline is data analysis to identify the expressed somatic mutations. All 
somatic mutations are subjected to a set of filters to exclude “false-positive” calls. The filters include: 
Exome data analysis:  
(1) Normal Coverage > 5x 
(2) Normal Variant Allele Frequency (VAF) < 2% 
(3) Tumor Coverage > 10x 
(4) Tumor Variant Allele Frequency > 20% 
 
Tumor cDNA Capture Data 
(5) Tumor Coverage > 10x 
(6) Tumor Variant Allele Frequency > 20% 
(7) FPKM > 1 

Washington University School of Medicine 
Protocol Date 08/03/23, A5 
Siteman Cancer Center 
Pancreatic Cancer SPORE 
Neoantigen DNA vaccines  
 
Page 21 of 69 
Pancreatic cancer-specific mutations that meet these filters will be prioritized. We will further prioritize 
indels over missense mutations. The mutations will be further analyzed for epitopes using the epitope 
prediction algorithms detailed below (Please see Section 5.4). 
Please see below for an explanation of the terms used above. 
The term coverage is a general term to describe the fold oversampling of a DNA target by sequencing 
data. In covering a target region or genome, increasing depth of coverage leads to increased certainty of 
variant detection. Therefore 10x “coverage” implies that the given site was independently sequenced at 
least 10 times. 
Variant allele frequency (VAF) is a metric that represents the sensitivity of identifying somatic variant over 
a range of sample purity and sequencing depths. The expected variant allele frequency acts as a 
surrogate for purity. For example, if a heterogeneous somatic variant is present in all tumor cells and the 
tumor cells represent 40% of the sample, then the observed VAF would be 20%. 
FPKM stands for Fragments Per Kilobase of transcript per Million mapped reads. This is a way to 
estimate expression of a gene. In RNA sequencing (and cDNA-capture sequencing), the relative 
expression of a transcript is proportional to the number of cDNA fragments that originate from it. Paired-
end RNA-Seq experiments produce two reads per fragment, but that doesn't necessarily mean that both 
reads will be mappable. For example, the second read is of poor quality. If we were to count reads rather 
than fragments, we might double-count some fragments but not others, leading to a skewed expression 
value. Thus, FPKM is calculated by counting fragments, not reads. If you have a gene with an FPKM of 0, 
you'll see a few reads that align to it. However, if there are 100 or more reads per sample, 1 or 2 reads is 
rather insignificant. So, while you can't really say the gene is not expressed with 100% certainty, you can 
say it was not detected.  
 
Epitope prediction algorithm 
We have developed and optimized an epitope prediction algorithm for the identification and prioritization 
of neoantigens. This optimized epitope prediction algorithm is described below and in the Gubin 2014 
manuscript [25]. We have established this algorithm is collaboration with Dr. Robert Schreiber, an 
internationally-known expert in tumor immunology [24, 55-62]. Schreiber was one of the first to use next 
generation sequencing technologies to identify neoantigens, demonstrating that these antigens are 
important tumor rejection antigens [24]. Schreiber and Gillanders have now optimized the epitope 
prediction algorithm and have demonstrated that cancer vaccines targeting neoantigens are associated 
with antitumor immunity [25].  
The goal of the optimized epitope prediction algorithm is to identify and prioritize up to 20 neoantigens. 
The algorithm uses a combination of binding algorithms, processing algorithms and in vitro binding 
assays. 
Mutations that are expressed in the pancreatic cancer will be identified using the sequencing pipeline 
outlined in the document titled “Sequencing Pipeline.” The predicted amino acid sequences 
corresponding to the expressed mutations will be pipelined through multiple class I and class II MHC 
epitope-binding algorithms provided by the Immune Epitope Database and Analysis Resource 
(http://www.immuneepitope.org) including but not limited to (i) Stabilized Matrix Method (SMM) [67], (ii) 
Artificial Neural Network (ANN) [68], and (iii) NetMHCpan [69]. 
A prioritized list of binding epitopes (i.e. IC50 < 500 nM) will be generated after calculating the median 
binding affinity value for each mutant sequence (affinity value expressed as 1/IC50 x 100).  
Filters will be applied to the list to (a) eliminate epitopes that are not processed efficiently by the 
immunoproteosome based on the NetChop algorithm [70] (peptides with a NetChop score > 0.6 will be 
prioritized), (b) deprioritize epitopes from hypothetical proteins or that form a weaker predicted binding 
epitope than that expressed by the corresponding wild type sequence, (c) prioritize mutant epitopes that 
have the highest difference in predicted binding affinity compared to their wild type counterpart, and (d) 
incorporate expression profiles of wild type genes that correspond to the mutant antigen candidates in 
normal vs tumor cells.  

Washington University School of Medicine 
Protocol Date 08/03/23, A5 
Siteman Cancer Center 
Pancreatic Cancer SPORE 
Neoantigen DNA vaccines  
 
Page 22 of 69 
To confirm the ability of prioritized neoantigens to bind to the corresponding MHC class I allele, we may 
synthesize peptides corresponding to the prioritized list of candidate epitopes. Synthetic peptides (> 95% 
purity) will be tested for binding to the respective class I MHC molecule using the T2 cell line, or one of its 
derivatives expressing common class I MHC alleles. The T2 cell line is deficient in transporters for antigen 
presentation (TAP), and as such is commonly used to assess binding of exogenously added peptides [17, 
71]. Expression of class I MHC alleles is measured by staining of T2 cells with antibody specific for the 
class I MHC allele followed by flow cytometric analysis to measure the fluorescence intensity of the class I 
MHC allele on the cell surface. Data are typically expressed as Mean Fluorescence Intensity (MFI) by 
subtracting the baseline fluorescence intensity of the class I MHC allele to that measured after addition of 
exogenous peptide. Neoantigens that do not bind to the corresponding class I MHC allele will be 
deprioritized. Similar algorithms will be used to predict class II neoantigens. 
 
Manufacturing facility 
The personalized polyepitope DNA vaccines will be prepared in the Siteman Cancer Center Biologic 
Therapy Core Facility at Washington University School of Medicine. The facility is located at 500 S. 
Kingshighway, Room 719 Southwest Tower, Saint Louis, MO 63110. 
The Biologic Therapy Core Facility Operations Director is Johnnie Cartwright. She has extensive 
experience in writing, implementing, and strictly adhering to phase appropriate cGMP procedures & 
regulations to provide for the highest level of product quality. 
The facility adheres to cGMP practices with regard to documentation, facility maintenance, and QC/QA 
review. Within the 2,615 sq ft GMP-facility on the 7th floor of the Southwest Tower, 6 manufacturing 
rooms are available for clinical grade manufacturing of cellular therapy products, recombinant DNA or 
gene therapy products (Floor plan below).  
 

Washington University School of Medicine 
Protocol Date 08/03/23, A5 
Siteman Cancer Center 
Pancreatic Cancer SPORE 
Neoantigen DNA vaccines  
 
Page 23 of 69 
 
In the Biologic Therapy Core facility, all manufacturing rooms are physically separated from each other 
with single pass air, and provide one-way personnel and product flow, which avoids cross-contamination 
of products. 6 different products can be worked on simultaneously without interfering with each other. 
Passthroughs are available from gowning rooms to intermediate rooms, and intermediate rooms to 
manufacturing rooms. All manufacturing rooms are completely isolated from each other by sealed walls 
and ceilings. A custom ceiling allows maintenance on the HVAC units, filters or lights without breeching 
the clean environment of the facility. 
The manufacturing laboratories are only entered, one at a time, from an intermediate room. The 
laboratories are only exited, one at a time, into another intermediate room. 
This provides easily manageable spatial and temporal segregation of products, which is a necessity to 
prevent product cross-contamination. Door interlocks, which allow for doors to be opened only one at a 
time, assure this unidirectional flow of personnel and products. 
Manufacturing rooms, intermediate rooms and gowning/degowning rooms are isolated from each other by 
air pressure gradients. The manufacturing rooms have higher air pressure in respect to the intermediate 
rooms; gowning rooms have negative air pressure towards intermediate rooms and the hallway. This also 
assures protection of the outside environment from potentially biohazardous materials produced in the 
Biologic Therapy Core facility. Air pressure gradients are maintained by individual HVAC units for each 
room. The manufacturing rooms are certified and maintained as Class 10,000, the intermediate rooms 
are certified and maintained as Class 100,000. 
Every manufacturing lab is equipped with a 6-foot biosafety cabinet and a dual chamber incubator. 
Sufficient room is available per manufacturing lab to accommodate specialized equipment (e.g. clinical 
grade magnetic bead cell separator) for different manufacturing processes. In the largest manufacturing 
room, besides the 6ft biosafety cabinet, a second 4ft biosafety cabinet is installed for work with infectious 
agents. Manufacturing room 1 can also be switched to negative air pressure and BSL-3 containment 
(vector manufacturing), and an IBC approved protocol to accomplish this task is in place. 
Quality control and monitoring plans are in effect. Daily environmental cleaning is performed by dedicated 
personnel. The facility is electronically monitored continuously, daily monitoring by core personnel 
ensures adherence to standards and is prescribed by SOPs. Environmental monitoring includes non 
viable and viable particle enumeration with limits more stringent than the standards prescribed in the 
USP. 
GMP regulations for even Phase 1 clinical trials are in place to assure a cellular or biological product to 
be safe and free of contaminants. 
 
 
 
 
 
 
 
 
Figure 3: Siteman Cancer Center Biologic Therapy Core Facility floor plan. HW: Hallway, G1: 
Gowning Room 1; G2: Gowning Room 2; IEN 1: Intermediate Entry Room 1; IEN 2: Intermediate Entry 
Room 2; M1 to M3: Manufacturing Rooms 1 to 3; M4 to M6: Manufacturing Rooms 4 to 6; IEX: 
Intermediate Exit Room; DG: Degowning Room; O: Office; A: Autoclave Room. 

Washington University School of Medicine 
Protocol Date 08/03/23, A5 
Siteman Cancer Center 
Pancreatic Cancer SPORE 
Neoantigen DNA vaccines  
 
Page 24 of 69 
 
Manufacturing process 
Overview of the polyepitope DNA vaccine manufacturing process. Schematic outline of product 
manufacture, in-process testing, and product release tests. 
 
 
Figure 4: Overview of the polyepitope DNA vaccine manufacturing process. Schematic outline of 
product manufacture, in-process testing, and product release tests. 

Washington University School of Medicine 
Protocol Date 08/03/23, A5 
Siteman Cancer Center 
Pancreatic Cancer SPORE 
Neoantigen DNA vaccines  
 
Page 25 of 69 
Design of the polyepitope DNA vaccine. Neoantigen DNA vaccines will be designed in the Gillanders 
laboratory at WUSM. Up to 20 candidate neoantigens and personalized mesothelin epitopes from each 
patient will be integrated into the polyepitope DNA vaccine. Neoantigens will be identified and prioritized 
based on the results of next-generation sequencing studies, epitope prediction algorithms, and in vitro 
studies. 
Synthesis of the polyepitope insert. A polyepitope insert integrating up to 20 of the highest priority 
neoantigens and personalized mesothelin epitopes will be designed in the Gillanders laboratory. This 
insert will then be synthesized by Blue Heron Biotech. The insert will then be cloned into the pING parent 
vector by Blue Heron Biotech. The pING vector containing the polyepitope insert will be sequenced by 
Blue Heron Biotech. The plasmid will then be used to establish a master cell bank at WUSM as detailed 
below. 
Creation of a master cell bank. Master cell banks for each individual polyepitope DNA vaccine will be 
constructed by transformation of E. coli strain DH5 alpha with each patient’s unique pING-polyepitope 
plasmid. Briefly, 15 vials of transformed bacteria will be prepared and stored at -80°C in the Siteman 
Cancer Center Biologic Therapy Core Facility. The Johnson Controls monitoring system continuously 
monitor the freezer used for storage.  The system triggers an alarm and electronically notifies staff when 
temperature values are out of specification. In addition, daily manual monitoring is applied. For each 
bacterial culture production, a vial of the glycerol stock will be taken from the master cell bank and used 
for culture inoculation. 
Samples from each master cell bank will be characterized for host strain identity, contaminating 
organisms, viable cells, plasmid activity, and plasmid identity as detailed below in testing to be performed 
under GLP conditions at Barnes-Jewish Hospital and Washington University School of Medicine. Of note, 
in previous sequencing reactions we have identified three variations in pING vector sequence. These 
variations are in non-coding areas of the plasmid DNA, and are not expected to have any functional 
significance. They are also present in the pING parent vector obtained from MSKCC, suggesting that the 
published sequence for the pING vector may not be accurate. Please see Table 1 below. 
TABLE 1: Master cell bank validation for polyepitope DNA vaccines 
Criteria 
Method 
Specification 
Test site 
Host strain identity 
Growth on 
selective media 
Growth of E. coli 
Barnes-Jewish Hospital 
Microbiology Laboratory 
Contaminating 
organisms 
Growth on 
selective media, 
No growth 
Barnes-Jewish Hospital 
Microbiology Laboratory 
Viable cells 
Growth in LB agar 
Growth of E. coli 
Barnes-Jewish Hospital 
Microbiology Laboratory 
Plasmid activity 
Transfect cells, 
RT-PCR 
Detection of polyepitope 
construct mRNA 
Gillanders Laboratory 
Plasmid identity 
DNA sequencing 
Sequence as predicted 
Gillanders Laboratory 
Plasmid identity 
Restriction 
analysis 
As expected, no 
extraneous bands 
Gillanders Laboratory 
Plasmid DNA fermentation. The plasmid vectors will be grown in E. Coli strain DH5alpha. The host 
bacteria will be grown in 500 mL LB broth cultures in a bacterial shaker at 37°C for 16 hours. Antibiotic 
selection (kanamycin) will be applied to increase plasmid yield.  
Plasmid DNA purification. The plasmid DNA will be purified using reagents obtained from Sigma Aldrich 

Washington University School of Medicine 
Protocol Date 08/03/23, A5 
Siteman Cancer Center 
Pancreatic Cancer SPORE 
Neoantigen DNA vaccines  
 
Page 26 of 69 
(Saint Louis, MO). Distinct lots of the Genelute HP Select Plasmid Gigaprep Kit from Sigma-Aldrich (St. 
Louis, MO) will be used for the manufacture of the plasmids. The following narrative is a brief description 
of the protocol. The composition of buffers, solutions, and chromatography material is included in Table 2 
below. (Please note that the precise composition of the resin and solutions are considered by Sigma 
Aldrich to be proprietary). The harvested bacteria will be recovered by centrifugation, and then 
resuspended in a cell suspension buffer containing RNase A. Lysis buffer will be added and a 
homogenous lysate will be obtained by gentle inversion. After neutralization of the bacterial lysate, a 
lysate clearing agent will be added and allowed to stand for 5 minutes for precipitates to form. The lysate 
will be cleared by filtration, and a binding solution optimized for endotoxin-free plasmid purification will be 
added. The lysate with binding solution will then run through an equilibrated column where the plasmid 
DNA is captured onto a silica membrane in the presence of high salt, while endotoxins are prevented 
from adsorbing to the membrane. The flow-through will be discarded, and the contaminants in the column 
will be removed in three wash steps. Finally, the bound plasmid DNA will be eluted in endotoxin-free 
water. The eluted DNA will be precipitated by adding 0.1 volume of 3 M sodium acetate and 0.7 volumes 
of isopropanol. The DNA will be recovered by centrifugation at 14,000 x g in a fixed angle rotor. A wash 
and pelleting of the final DNA with 70% ethanol will be performed and the resulting purified DNA will be 
air dried in a biosafety cabinet. All vessels and buffers used for DNA preparation will be sterile for single 
use, or will be autoclaved in a monitored and quality controlled autoclave in the Biological Therapy Core 
facility. 
The only product of animal origin is the RNase solution. This is isolated from bovine pancreas and we 
have Certificates of Origin that clearly indicate that these were derived from a New Zealand herd. These 
animals passed ante and post mortem testing for viral pathogens; the samples were treated with low pH 
(1.7) for 12 hours to inactivate pathogenic viral agents; and no animal protein was fed to these animals.  
Glycerol will be used in our manufacturing process. However, all Lot #s of the glycerol utilized will be of 
plant origin. 
 

Washington University School of Medicine 
Protocol Date 08/03/23, A5 
Siteman Cancer Center 
Pancreatic Cancer SPORE 
Neoantigen DNA vaccines  
 
Page 27 of 69 
TABLE 2: Composition of buffers, solutions, and chromatography material 
Solution 
Composition, Sigma # 
Wash Solution 2 
Tris-HCl (T3038), NaCl (S5150) 
Neutralization Solution 
Trizma-HCl (T5941) 
Binding Column 
Guanidine Thiocyanate (G9277), EDTA (E7889) 
Column Preparation 
Solution 
NaOH (S2770) 
Resuspension/RNase A 
Solution 
Tris-HCl (T3038), EDTA (E7889) 
Lysis Solution 
NaCl (S5150), Tris-HCl (T3038), EDTA (E7889), Ocytl-beta-
Thioglucopyranoside (O6004) 
Lysate Clearing Agent 
Guanadine HCl (G3272), Tris-HCl (T2413), EDTA (E7889), Tween20 
(P7949), Triton X-100 (T8787), Hydrochloric acid (H9892) 
Binding Solution 
Guanadine HCl (G3272), Tris Acetate (T1258), Isopropanol (I0398), 
NaOH (S2770)  
Column Prep Solution 
NaOH (S2770) 
EndoCleaning Solution 
NaOH (S2770), Tween20 (P7949) 
Wash Solution 1 
NaCl (S5150), Tris-HCl (T3038) 
Wash Solution 2 
Tris-HCl (T3038), NaCl (S5150) 
 
Product formulation 
The eluted plasmid preparations will be resuspended in sterile, preservative free-PBS. A small sample 
(100 µL) will be removed aseptically and the concentration tested using a certified NanoDrop 
spectrophotometer, Qubit Fluorometer or similar device. Typically, the eluted DNA is in the range of 3-4 
mg/mL. The DNA product will be diluted to 2 mg/mL using sterile, preservative-free PBS, and aseptically 
added to 2.0 mL cryovials from Nalgene (Catalog# 5000-0020, Lot# 616412). The vials are made of 
polypropylene with a high-density polyethylene closure. They are externally-threaded for aseptic 
technique. They have a sealing ring in conjunction with specially designed threads. A white marking area, 
fill line, and graduations are printed on the vials with white paint. They are radiation-sterilized, non-
cytotoxic, non-pyrogenic, and are RNase/DNase-free. 
We plan to synthesize enough DNA for approximately 20 vials. We anticipate that 12 vials will be required 
for vaccination (2 vials per time point times 6 timepoints), and 5 vials will be required for product release 
tests. We are targeting 20 vials to have a reserve stock. 
Following vialing, we anticipate that 4 vials (#1, 4, 10, 19) will be used for product release testing. Vials 1 
and 19 will be used to test sterility and represent the beginning and end of the vialing procedure. Two 
additional vials will be used for the other product release tests listed in Table 5. 
If any contamination would be detected in our plasmid samples, we would initiate an alcohol precipitation 
step. This procedure is basically the final stage of the gigaprep purification, where the soluble DNA is 
precipitated using 2x volumes of cold isopropanol and the DNA is pelleted by centrifugation. The pellet is 

Washington University School of Medicine 
Protocol Date 08/03/23, A5 
Siteman Cancer Center 
Pancreatic Cancer SPORE 
Neoantigen DNA vaccines  
 
Page 28 of 69 
washed in 70% ethanol and air dried. This procedure is a standard method for eliminating any 
microbiologic contamination in DNA. 
 
Product release tests 
A key issue in the quality control of plasmid DNA for human use is the development of analytical 
methodologies capable of fully characterizing the product in its final form, ensuring the production of a 
consistent product. The following list of release criteria, analytical methodologies, and specifications has 
been adapted from [72]. Polyepitope DNA vaccines will be tested for the following specifications. 
TABLE 3: Polyepitope DNA vaccine product release tests 
Criteria 
Method 
Specification 
Test Site 
Plasmid activity 
Transfect cells, RT-PCR 
Polyepitope mRNA 
present 
Gillanders Laboratory 
Residual kanamycin 
USP 29 <81> 
< 8 mcg/mL 
Alcami or equivalent 
Plasmid 
homogeneity 
Agarose gel 
electrophoresis 
≥80% closed circular 
Gillanders Laboratory 
Residual RNA 
Agarose gel 
electrophoresis 
Less than 1% 
Gillanders Laboratory 
Residual bacterial 
chromosomal DNA 
Agarose gel 
electrophoresis 
Less than 1% 
Gillanders Laboratory 
Plasmid identity 
Restriction analysis 
As expected, no 
extraneous bands 
Gillanders Laboratory 
Appearance 
Visual 
Clear and colorless, 
free of particulates 
Gillanders Laboratory 
pH 
Calibrated pH meter 
Report pH 
Biologic Therapy Core 
Facility 
DNA concentration 
Nano-drop 
spectrophotometer 
Greater than 1 
mg/mL 
Gillanders Laboratory 
Residual proteins 
BCA reagent colorimetric 
Less than 1% (< 21 
µg/mL) 
Gillanders Laboratory, 
Biologic Therapy Core 
Facility, or equivalent 
Residual 
isopropanol, EtOH 
USP <467> 
Less than 5000 ppm 
PACE Analytics or 
equivalent 
Endotoxin 
Chromogenic Limulus 
amebocyte lysate assay 
Less than 175 
EU/mL 
Biologic Therapy Core 
Facility 
Sterility 
USP <71> 
No growth 
Biologic Therapy Core 
Facility 
Alternatively, we will perform studies to demonstrate that the manufacturing process removes residual 
kanamycin, RNA, bacterial chromosomal DNA, proteins, isopropanol and EtOH below the specification 
levels, and not perform these residual studies on each lot. To date, testing on each lot has shown that 
residual kanamycin is undetectable and residual isopropanol and EtOH is consistently below the 
specification. This testing will no longer be performed. The criteria for sterility meets specification if no 
growth after 7 days. The sterility culture will be held and monitor for 14 days as per USP <71>. 

Washington University School of Medicine 
Protocol Date 08/03/23, A5 
Siteman Cancer Center 
Pancreatic Cancer SPORE 
Neoantigen DNA vaccines  
 
Page 29 of 69 
 
Stability 
The polyepitope DNA vaccines will be stored in the Biologic Therapy Core Facility in a -80°C freezer. The 
freezer used for storage is monitored 24/7 by Johnson Controls monitoring system with dedicated on-call 
personnel to address any aberrant occurrences. Overall, we anticipate no problems with plasmid stability 
as stability is considered to be one of the advantages of the DNA vaccine platform, and plasmid DNA 
formulated in PBS is considered to be very stable. 
We are currently storing similar investigational DNA vaccines in the Biologic Therapy Core Facility in a -
80°C freezer. These investigational DNA vaccines were manufactured in the Biologic Therapy Core 
Facility using an almost identical manufacturing process as the one proposed here, are based on the 
pING parental vector and are thus very similar in size and composition, and are formulated similarly in 
PBS at 2 mg/mL. These DNA vaccines have remained stable over a 3 year period.  
We do not anticipate that long-term stability testing will be required. As each lot of vaccine is personalized 
to a specific patient, we anticipate that the vaccine will be used within 6 months of completion of the 
product release tests. 
Vaccines to be administered at other participating institutions will be shipped on dry ice to the address 
provided by the institution. 
 Labels 
Each polyepitope DNA vaccine product will be made as a single lot ensuring that the labeled product is 
unique and consistent for this trial. Please see the sample label below. Prior to administration a time out 
will be performed to confirm subject identity, and the investigational agent to be administered. This will be 
performed by the clinical research associate, the oncology nurse trained to administer the vaccine by 
electroporation and the experimental pharmacist. 
SCC Biologic Therapy Core Facility 
Saint Louis, MO 
Store at -80°C 
WUSM PPDV #1 
Subject Study #, Subject Initials 
LOT # 
1 
Vial # 
003 
 
Concentration 
2mg/mL 
Volume 
1.2mL 
Date 
mm/dd/yyyy 
CAUTION Investigational drug limited by federal law to investigational use 
 Investigational agent administration 
Each DNA vaccination will be 1 mL vaccine administered intramuscularly using an integrated 
electroporation administration system (TDS-IM system, Ichor Medical Systems). At each vaccination time 
point, patients will receive two injections at separate sites. 
5.11.1 Introduction to electroporation 
Electroporation (EP) is a potent delivery technique based on the in vivo application of electrical fields that 
may improve immune responses following the administration of plasmid DNA vaccines. The EP effect is 
induced through the propagation of electrical fields of sufficient magnitude and duration within a target 
region of tissue. These electrical fields produce a transient increase in the membrane permeability of cells 
exposed to the electrical field, allowing enhanced intracellular uptake of agents distributed within the 
interstitium of the local tissue. Of note, agent delivery occurs only where the agent of interest is present 
contemporaneously with the propagation of threshold level electrical fields. Numerous non-clinical studies 

Washington University School of Medicine 
Protocol Date 08/03/23, A5 
Siteman Cancer Center 
Pancreatic Cancer SPORE 
Neoantigen DNA vaccines  
 
Page 30 of 69 
have demonstrated that the application of EP can enhance gene expression and immune responses 
following plasmid DNA vaccine delivery. 
Although DNA vaccines encoding antigens from a variety of cancers and infectious pathogens have 
exhibited considerable promise in non-clinical studies, clinical experience to date suggests that current 
administration methods may have insufficient potency to achieve the desired immune responses in 
humans. Poor delivery efficiency and low levels of antigen expression have been implicated as factors 
contributing to the sub-optimal responses observed to date [73, 74]. 
Electroporation (EP) is a potent physical delivery technique based on the in vivo application of electrical 
fields. The propagation of electrical fields of a sufficient magnitude within a target tissue induces a 
transient increase in cell membrane permeability, resulting in a significant improvement of intracellular 
uptake of exogenous substances. Shortly after pulse delivery, the cell membrane function stabilizes and 
cells within the affected tissue resume normal function. EP has been shown in non-clinical studies to be a 
potent method for delivery of DNA vaccines [47, 75-77] 
EP has several important advantages that make it an appealing method for delivery of DNA vaccines. 
First, EP can reliably enhance delivery and expression of DNA antigens by 2-3 orders of magnitude. This 
improvement in delivery has been shown to enhance both cellular and humoral immune responses. 
Second, EP is a non-viral delivery method. As such, it has a favorable safety profile and does not elicit 
unwanted immune responses against the vector. Thus, the technique is particularly appropriate for 
indications likely to require multiple vaccine administrations to achieve response (e.g. prime / boost). It is 
also advantageous because the absence of immunogenic viral antigens minimizes potentially deleterious 
antigen competition, thereby ensuring that the immune response is concentrated on the antigen of 
interest. Lastly, unlike other non-viral approaches, EP is capable of inducing consistent responses from 
subject to subject, even at relatively low DNA doses. 
5.11.2 The TDS-IM Electrode Array System 
The TDS-IM device is supplied by Ichor Medical Systems, Inc. The TDS-IM device utilizes the in vivo 
application of electrical fields to enhance the intracellular delivery of agents of interest in a targeted region 
of tissue (EP). The device complies with the applicable safety and electromagnetic compatibility 
requirements of International Electrotechnical Commission (IEC) 60601-1. During this study, the device 
will be operated according to the TDS-IM User Manual and applicable study-specific procedures to 
ensure consistent and safe utilization. 
Each TDS device consists of 3 parts: A pulse stimulator, an integrated applicator, and a single-use 
application cartridge (see Figure below). The components are manufactured in ISO13485-compliant, 
FDA-registered facilities including BIT MedTech, LLC. (Pulse Stimulator) and Life Science Outsourcing, 
Inc. (Integrated Applicator and Application Cartridges. 
 
5.11.3 Summary of TDS-IM Clinical Experience to Date 
To date, formal safety studies have been conducted to assess the TDS-IM as the means for delivery of 
nine DNA vaccine candidates. This includes studies of a melanoma DNA vaccine encoding a xenogeneic 
form of the tyrosinase antigen that uses the same vector backbone (pING) as the Mammaglobin-A DNA 
vaccine candidate (see Table 4 & Table 5). Safety studies for the various DNA vaccine candidates have 
included repeat dose studies in rabbits to assess safety and toxicology, and studies in rats to assess DNA 
vaccine biodistribution, persistence, and potential for integration into host DNA. 
TDS-IM Components
TDS-IM Integrated 
Applicator
TDS-IM Application 
Cartridge
TDS Pulse Stimulator
Syringe
TDS-IM Components
TDS-IM Integrated 
Applicator
TDS-IM Application 
Cartridge
TDS Pulse Stimulator
Syringe

Washington University School of Medicine 
Protocol Date 08/03/23, A5 
Siteman Cancer Center 
Pancreatic Cancer SPORE 
Neoantigen DNA vaccines  
 
Page 31 of 69 
In the safety and toxicology studies, notable findings associated with electroporation-mediated delivery of 
DNA vaccines were limited to localized inflammatory responses of mild to moderate severity at the site of 
administration (for an example, see Dolter, K.E., et al. Immunogenicity, safety, biodistribution and 
persistence of ADVAX, a prophylactic DNA vaccine for HIV-1, delivered by in vivo electroporation. 
Vaccine, 2011. 29(4): p. 795-803). The injection site findings were most prominent in tissue samples 
obtained 48–72 hours after administration at that site. Tissue samples obtained from administration sites 
14–43 days after administration indicate progressive resolution of the local inflammatory responses over 
time. 
The results of the biodistribution studies indicated negligible systemic uptake of vaccine DNA following 
EP-based intramuscular delivery, with no significant differences observed among the different DNA 
vaccine candidates tested to date. Persistence analysis indicated that the presence of vaccine DNA 30 to 
90 days after administration was confined to the tissues at the site of administration (muscle and skin), 
and only at very low levels (< 1,000 copies/µg host genomic DNA), suggesting minimal risk for potential 
integration of vaccine DNA into host DNA (for an example, see Dolter, 2011). 
Reports describing the results of the tyrosinase GLP safety and toxicology studies are available in BB 
IND 13275; a letter of cross reference to this IND has been provided by Ichor. 
5.11.4 Summary of TDS-IM clinical experience to date 
The TDS-IM investigational device is currently being used as the means for DNA vaccine delivery in both 
the therapeutic and prophylactic setting. Six clinical trials of the TDS-IM device have been completed as 
detailed in the Table below. 
TABLE 4: TDS-IM Based DNA Vaccine Administration: Completed Clinical Trials 
ClinicalTrials.gov 
Study # 
Vaccine Candidate 
Subject 
Population 
NCT00545987 
Multigenic HIV-1 DNA vaccine candidate (ADVAX) 
Healthy, HIV 
uninfected adult 
volunteers 
NCT00471133 
Xenogeneic tyrosinase DNA vaccine candidate 
(pINGmuTyr) 
Patients with 
Stage IIB-IV 
melanoma 
NCT01169077 
Multi-epitope malaria DNA vaccine (EP-1300) 
Healthy adult 
volunteers 
NCT01502345 
Multi-antigen hantavirus DNA vaccine (pWRG/HTN-M(x) 
and pWRG/PUUV-M(s2)) 
Healthy adult 
volunteers 
NCT01641536 
Multi-antigen HBV DNA vaccine administered with a DNA-
based human IL-12 adjuvant (HB-110) 
HBV infected adult 
volunteers 
NCT01496989 
Multi-antigen HIV DNA vaccine (HIV-MAG) administered 
with or without a DNA-based human IL-12 adjuvant 
(GENEVAX) prior to or after administration of an 
adenovirus vector (Ad35GRIN) 
Healthy, HIV 
uninfected adult 
volunteers 
The TDS-IM device is currently being evaluated in seven ongoing clinical studies as detailed in the Table 
below. 
TABLE 5: TDS-IM Based DNA Vaccine Administration: Ongoing Clinical Trials 
ClinicalTrials.gov 
Study # 
Vaccine Candidate 
Subject Population 
NCT01138410 
Epitope-based TRP-2 melanoma vaccine (SCIB-1) 
Patients that are HLA A1, 
A2, A24, or B35 positive 
with AJCC stage III-IV 
melanoma 
NCT01859325 
Multi-antigen HIV DNA vaccine (HIV-MAG) 
administered with or without a DNA-based human 
IL-12 adjuvant (GENEVAX) prior to administration 
of an vesicular stomatitis virus vector (rVSVgag) 
HIV infected adult 
volunteers 

Washington University School of Medicine 
Protocol Date 08/03/23, A5 
Siteman Cancer Center 
Pancreatic Cancer SPORE 
Neoantigen DNA vaccines  
 
Page 32 of 69 
NCT01578889 
Multi-antigen HIV DNA vaccine (HIV-MAG) 
administered with or without a DNA-based human 
IL-12 adjuvant (GENEVAX) prior to administration 
of an vesicular stomatitis virus vector (rVSVgag) 
Healthy adult volunteers 
NCT01266616 
Multi-antigen HIV DNA vaccine (HIV-MAG) 
administered with or without a DNA-based human 
IL-12 adjuvant (GENEVAX) 
HIV infected adult 
volunteers 
NCT01634503 
Multi-antigen HPV DNA vaccine administered with 
a DNA-based human IL-12 adjuvant (GX-188E) 
Patients with Grade 3 
Cervical Intraepithelial 
Neoplasia 
NCT01493154 
HPV DNA vaccine administered with a DNA-based 
calreticulin adjuvant (pNGVL-4a-CRT/E7 (detox)) 
HPV-associated squamous 
cell carcinoma of the head 
and neck 
NCT01984983 
VEEV DNA Vaccine Candidate 
Healthy adult volunteers 
To date, the 13 clinical trials that have been completed or are currently ongoing have enrolled over 350 
subjects in the electroporation arms of the studies (including subjects receiving either the DNA vaccine 
candidate or placebo). The device has been used for administration of DNA injections of up to 1.0 ml 
volume and 4.0 mg DNA dose per injection site. Subjects have received the vaccine candidate either as a 
single injection in one muscle site (total DNA dose up to 4.0 mg per administration time point) or as 2 
injections in 2 separate muscle sites (total DNA dose up to 8.0 mg per administration time point). Subjects 
given the DNA dose as a single injection have received up to 5 TDS-IM injections at up to 4.0 mg DNA, 
and subjects administered the DNA dose in 2 injections have received up to 5 administrations (i.e., 10 
total TDS-IM injections).  
Adverse responses reported in association with use of the device include localized muscle contractions 
and associated discomfort/pain during the application of EP, minor cutaneous bleeding at the site of 
injection, and transient injection site soreness of mild to moderate severity, typically resolving within 24–
72 hours following administration. Several subjects in the 2 melanoma studies have reported 
lightheadedness immediately following procedure application which, in some cases, was accompanied by 
a decrease in blood pressure. In one subject, enrolled in the xenogeneic tyrosinase study, this was 
followed by a brief syncopal episode (~30 seconds duration) shortly after procedure application. The 
subject recovered without incident. At the time of enrollment, the subject indicated a life long history of 
sinus bradycardia of unknown origin, which was confirmed by electrocardiogram (EKG) during screening. 
Multiple EKGs performed after the syncopal episode indicated no changes from pre-procedure baseline. 
Based on the judgment of the investigator, the subject was withdrawn from the study and the study 
eligibility criteria were modified to exclude subjects with sinus bradycardia. No other serious or 
unanticipated adverse events attributed to the device or administration procedure have been observed. 
The results of the completed HIV-1 ADVAX vaccine study in healthy subjects have been published 
(Vasan S et al. In vivo electroporation enhances the immunogenicity of an HIV-1 DNA vaccine candidate 
in healthy volunteers. PLoS One 2011;6(5):e19252). Briefly, results from this study indicate that EP-
based delivery with the TDS-IM device at ADVAX DNA doses ranging from 0.2–4.0 mg was safe and 
effective in improving the magnitude, breadth and durability of cellular immune responses to a DNA 
vaccine candidate. Assessment of the tolerability of the EP procedure by questionnaire after each 
administration indicates that the procedure is generally acceptable for use in healthy subjects. 
5.11.5 Preparation of the vaccine 
Doses of the neoantigen DNA vaccines will be prepared in the Siteman Cancer Center Investigational 
Pharmacy/JHU equivalent pharmacy and delivered to the clinic on the day of vaccination. Vials will be 
thawed at room temperature on the day of study visits. After thawing the vaccine vial(s), the 
investigational pharmacist will mix the vial(s) completely by inverting the vial(s) at least 10 times (one 
inversion = one 180° turn of the wrist and back). The vaccine will then be withdrawn into 3.0 mL Becton 
Dickinson Model 309585 syringe under sterile conditions by the research pharmacist. For administration, 
a 22 gauge 1.5 inch injection needle will be affixed to the syringe. Once the dose is prepared, the 
research pharmacist will load the syringe into a TDS-IM Application Cartridge in a manner consistent with 
the instructions provided in the TDS-IM User’s Guide.  

Washington University School of Medicine 
Protocol Date 08/03/23, A5 
Siteman Cancer Center 
Pancreatic Cancer SPORE 
Neoantigen DNA vaccines  
 
Page 33 of 69 
5.11.6 Vaccine administration 
Detailed instructions for procedure administration using the TDS-IM device are included in the TDS-IM 
User’s Guide. Briefly, the TDS-IM Application Cartridge is loaded into the Integrated Applicator. The Pulse 
Stimulator is connected to an appropriate power source, turned on, and then connected to the Integrated 
Applicator through the supplied cable. Prior to administration, the skin at the site of administration is 
prepared according to the standard procedures for a conventional intramuscular injection. Once the skin 
has been disinfected, the skin around the injection site is held firmly while the Application Cartridge is 
placed against the injection site at a 90° angle. An indicator light on the device notifies the user that 
sufficient pressure has been applied. The activation button is then depressed, causing the electrodes and 
injection needle to be inserted into the target tissue. An automated safety check is performed, and, if 
passed, allows the injection of the study vaccine into the muscle. The procedure will conclude with a 
series of brief, localized muscle contractions at the administration site. The procedure will require 
approximately 7-10 seconds to complete, during which time the site is held firmly and with the device 
depressed against the skin. Once the indicator light on the device indicates that the procedure is 
complete, the device is withdrawn and site is covered with a sterile covering and pressure applied with 3 
fingers for 1 minute. 
 Investigational agent accountability 
5.12.1 Documentation 
The investigational agent will be prepared by the study pharmacist at the Siteman Cancer Center 
Investigational Pharmacy/JHU equivalent pharmacy. The study pharmacist will be responsible for 
maintaining an accurate record of the codes, inventory, and an accountability record of vaccine supplies 
for this study. Electronic documentation as well as paper copies will be used. 
5.12.2 Disposition 
The empty vials and the unused portion of a vial will be discarded in a biohazard containment bag and 
incinerated or autoclaved. Any unopened vials that remain at the end of the study will be returned to the 
production facility or discarded at the discretion of the principal investigator in accordance with policies 
that apply to investigational agents. Partially used vials will not be administered to other subjects or used 
for in vitro experimental studies. They will be disposed of in accordance with institutional or pharmacy 
policy. 
 
6 
TREATMENT PLAN 
 
Plasmid DNA administration 
All subjects will be treated as outpatients in the Siteman Cancer Center/Sidney Kimmel Comprehensive 
Cancer Center. 
Patients with pathologically verified R0/R1surgical resection and no evidence of recurrent and/or 
metastatic disease will be identified for tissue procurement and sequencing, and patients will be 
consented within the 16 weeks following surgical resection to allow time for vaccine creation. Patients will 
initiate gemcitabine/capecitabine, mFOLFIRINOX or comparable adjuvant chemotherapy no more than 12 
weeks following resection. Adjuvant chemotherapy will be administered as per the NCCN 
recommendations for adjuvant chemotherapy following surgical resection. Additional chemoradiation, 
dose modifications, reductions, and supportive care will be performed at the discretion of the treating 
medical oncologist. After completion of adjuvant chemotherapy, patients will undergo repeat imaging by 
CT scan and/or MRI to evaluate for disease recurrence. Patients with no evidence of disease as 
determined by a board certified radiologist and without rising levels of tumor markers will be eligible for 
vaccination. If vaccine production is unable to be completed for a particular patient, that patient will 
discontinue participation in this study and will be replaced. 
The schedule for vaccination will be weeks 1, 5, 9, 13, 17, and 21. All vaccines will occur within +/- 1 
week with at least 3 weeks between vaccines. The first vaccine dose may be administered following 
confirmation of disease-free status. All study injections will be given intramuscularly using an integrated 

Washington University School of Medicine 
Protocol Date 08/03/23, A5 
Siteman Cancer Center 
Pancreatic Cancer SPORE 
Neoantigen DNA vaccines  
 
Page 34 of 69 
electroporation device (TDS-IM system, Ichor Medical Systems) by a trained healthcare provider and per 
the manufacturer’s instructions. At each vaccination time point, patients will receive two injections of the 
neoantigen DNA vaccine, one injection into each deltoid or lateralis. If both injections cannot be given on 
the same day, the patient will be asked to return to complete the second injection. Standard aseptic 
technique and precautions will be utilized in site preparation, vaccine administration, and medical waste 
disposal to ensure maximal safety of subjects and study personnel. 
At the discretion of the treating physician, patients may be pre-medicated with lorazepam 1mg PO at least 
30 minutes but no greater than 60 minutes prior to the first injection. Patients may also receive a second 
dose of lorazepam (1 mg PO) 10 minutes prior to injection. Patients may also receive acetaminophen 
650mg PO at least 30 minutes but no greater than 60 minutes prior to the first injection. 
The propagation of electroporation inducing electrical fields in the muscle will result in brief, localized 
muscle contractions at the site of administration, which are transiently painful. The neoantigen DNA 
vaccines will be administered by an experienced nurse who has completed a training seminar on the use 
of the TDS-IM device. The sites of immunization will be rotated for each of the immunizations.  
Following study injections, subjects will be observed for a minimum of 30 minutes. Vital signs 
(temperature, blood pressure, pulse and respiratory rate) will be taken at 30-45 minutes post-
immunization. The injection sites will be inspected for evidence of local reaction. 
On each injection day (prior to injection), study subjects will be evaluated by clinical exam and laboratory 
tests. Post-vaccination follow-up visits are at Week 25 ± 7 days and Week 73 ± 14 days. Additional follow-
up visits or telephone contact will be scheduled annually thereafter if the patient is alive and available for 
follow-up. 
At intervals throughout the study (both before and after vaccination) subjects will have blood drawn for 
immunologic assays. Any cells, serum or plasma not used will be stored for future immunological assays. 
Please see Section 8 Study Calendar for details on study visit procedures and monitoring. 
 
Definitions of Evaluability 
All enrolling patients are evaluable for the primary objective of feasibility, even patients whose 
tumor/normal exome sequencing is not completed or who are unable to have vaccine manufactured or 
administered. 
A patient must have received at least one injection of vaccine in order to be evaluable for the primary 
objective of safety and the secondary objective (prevalence of antigen-specific T cells in peripheral 
blood). 
 
Duration of therapy 
In the absence of treatment delays due to adverse events, treatment may continue for 6 doses of 
neoantigen DNA pancreatic cancer vaccines. Under certain circumstances, a subject will be terminated 
from participating in further injections. Subjects who are discontinued from additional study injections will 
continue to be followed according to the schedule of safety and immunogenicity evaluations. Please see 
Section 10 Removal of Patients from Protocol Therapy for additional details. 
 
Duration of follow up 
Patients will examined at 4 and 52 weeks following the final vaccination (Weeks 25 and 73). Additional 
follow-up visits or telephone contact will be scheduled every year thereafter if the patient is alive and 
available for follow-up. Patients removed from study for unacceptable adverse events will be followed 
until resolution or stabilization of the adverse event. 
 

Washington University School of Medicine 
Protocol Date 08/03/23, A5 
Siteman Cancer Center 
Pancreatic Cancer SPORE 
Neoantigen DNA vaccines  
 
Page 35 of 69 
7 
POTENTIAL TOXICITY AND DOSE MODIFICATIONS 
 
Previous human experience 
7.1.1 
Experience with the pING parent vector 
The neoantigen DNA vaccines have not been used in humans to date. Conventional DNA cancer 
vaccines based on the pING parent vector have been used extensively in phase 1 human clinical trials. 
We have discussed this issue with Dr. Robert Jambou at the Office of Biotechnology Activities, National 
Institutes of Health. On January 12, 2009, we made a Freedom of Information Act (FOIA) request for 
copies of information on the study population, dosing regimen, and study results to include a detailed 
adverse events/safety information for six NIH OBA-registered clinical trials: 0005-394, 0105-474, 0105-
474, 0303-573, 0312-617, 0412-684 and 0412-685. Dr. Jambou sent us the relevant clinical protocols, 
Appendix M documents, safety reports and annual reports. 
In accordance with Appendix M-I-C-4 of the NIH Guidelines for Research Involving Recombinant DNA 
Molecules posted April 1, 2002 in the Federal Register, investigators who have received authorization 
from the FDA to initiate a human gene transfer protocol must report in writing any unanticipated problems 
(serious adverse events or SAEs) to the NIH Office of Biotechnology Activities (OBA). These guidelines 
state that "Principal Investigators must submit, in accordance with Appendix M-I-C-4, a written report on 
any serious adverse event that is both unexpected and associated with the use of the gene transfer 
product. Any serious adverse event that is fatal or life-threatening, that is unexpected, and associated 
with the use of the gene transfer product must be reported to the NIH OBA as soon as possible, but not 
later 24 hours after the sponsor’s initial receipt of the information. Serious adverse events that are 
unexpected and associated with the use of the gene transfer product, but are not fatal or life-threatening, 
must be reported to the NIH OBA as soon as possible, but not later than 15 calendar days after the 
sponsor’s initial receipt of the information."  
We have used this information to prepare Table 6 below. Of note, there was only one serious adverse 
event reported. One patient with metastatic prostate cancer developed urinary retention. It is not clear if 
this was related to the DNA vaccine. Review of the available annual reports confirms minimal toxicity 
associated with DNA vaccination in these trials. Over 90% of the toxicities related to treatment were 
grade 1 and included diarrhea, dizziness, edema, fatigue, injection site reaction, nausea, vomiting, and 
rigors.  
 

Washington University School of Medicine 
Protocol Date 08/03/23, A5 
Siteman Cancer Center 
Pancreatic Cancer SPORE 
Neoantigen DNA vaccines  
 
Page 36 of 69 
Table 6: Clinical Experience with pING parent vector 
Protocol 
number  
Study 
population  
Dosing 
regimen  
Route of 
administration  
Number and 
frequency  
Safety  
0005-394  
Metastatic 
melanoma, 18 
patients total  
100, 500, or 
1500 ug 
DNA  
Intramuscularly 
using Biojector 
2000  
three 
vaccinations 
three weeks 
apart  
No SAEs 
reported  
0105-474  
Prostate 
cancer, 36 
patients  
100, 500, or 
1500 ug 
DNA  
Intramuscularly 
using Biojector 
2000  
six vaccinations 
three weeks 
apart  
One SAE 
reported, 
deemed to be 
unrelated to the 
vaccine  
0303-573  
Metastatic 
melanoma, 18 
patients total  
100, 500, or 
1500 ug 
DNA  
Intramuscularly 
using Biojector 
2000  
six vaccinations 
three weeks 
apart  
No SAEs 
reported  
0312-617  
Renal Cell 
Carcinoma, 18 
patients  
500, 1500, 
or 3000 ug 
DNA  
Intramuscularly 
using Biojector 
2000  
six vaccinations 
three weeks 
apart  
No SAEs 
reported  
0412-684  
Breast cancer, 
12 patients  
500, 1000, 
3000, or 
6000 ug 
DNA  
Intramuscularly 
using Biojector 
2000  
five 
vaccinations 
three weeks 
apart  
No SAEs 
reported  
0412-685  
Metastatic 
melanoma, 18 
patients  
500, 2000, 
or 4000 ug 
DNA  
Intramuscularly 
using Biojector 
2000  
six vaccinations 
three weeks 
apart  
No SAEs 
reported  
 
Potential toxicity 
7.2.1 
Potential toxicity related to the neoantigen DNA vaccine 
The primary objective of this trial is to evaluate the safety of the neoantigen DNA vaccines. This is one of 
the first times that neoantigens identified by next generation sequencing have been targeted for immune 
therapy in humans. This is also the first time that neoantigen DNA vaccines have been administered to 
humans. However, clinical trials of similar investigational plasmid DNA vaccines suggest that these 
vaccines will be very safe. We expect that most of the toxicity to be limited to local grade 1 or 2 reactions 
at the vaccination site. 
Please note that the risks detailed below are based on the risks of injections, the risks of vaccines in 
general, and the results of previous studies with investigational DNA vaccines. 
Risks associated with intramuscular administration of a DNA plasmid include acute bleeding and/or 
bruising. Although highly unlikely, intramuscular injection can result in muscle damage, peripheral nerve 
damage, and/or injection site infection. Due to the insertion and activation of multiple electrodes, use of 
the TDS-IM electroporation device may increase these risks. The propagation of electroporation inducing 
electrical fields in the muscle will result in brief, localized muscle contractions at the site of administration, 
which are transiently painful. As with any immunization, discomfort or redness at the injection site in the 
days following DNA vaccine administration may be expected. Since intramuscular DNA delivery with 
electroporation results in increased intracellular uptake of plasmid at the site of injection and electric field 
application, the procedure may increase the frequency and/or severity of local site reactions compared to 
conventional intramuscular administration of DNA vaccines. Such symptoms should not last longer than 
several days.  

Washington University School of Medicine 
Protocol Date 08/03/23, A5 
Siteman Cancer Center 
Pancreatic Cancer SPORE 
Neoantigen DNA vaccines  
 
Page 37 of 69 
Study subjects can receive medications such as acetaminophen, NSAIDs, or antihistamines as required. 
Steroids will not routinely be used in study subjects; if steroids are required the study subject will receive 
no further immunizations, but will continue to be monitored in follow-up visits. However, topical, inhaled, 
or locally injected steroids are acceptable on trial. 
Subjects may exhibit general signs and symptoms associated with administration of a vaccine injection, 
including fever, chills, rash, aches and pains, nausea, headache, dizziness and fatigue. These side 
effects will be monitored, but are generally short term and do not require treatment. 
The possibility of integration of the DNA plasmid vector into genomic DNA of transfected myocytes has 
been considered. Plasmid integration at a sufficiently high frequency carries the possibility of inducing 
deleterious mutations. Potential side effects could include an increased risk of malignancy arising from 
the cells harboring the mutation(s). However, current evidence from laboratory and animal studies 
indicates that the frequency of induced mutations following electroporation of DNA is conservatively 
estimated to be two to three orders of magnitude lower than that of naturally-occurring gene inactivating 
mutations in healthy humans. 
The effect of this vaccine on a fetus or nursing baby is unknown, so female subjects of child bearing 
potential will be required to agree to use birth control for sexual intercourse beginning 21 days prior to 
enrollment and continuing through the last protocol visit. Women who are pregnant or nursing will be 
excluded from the study. 
The potential discomforts of this study include having blood drawn, intramuscular injection of the vaccine, 
and possible reactions to the vaccine. Drawing blood causes transient discomfort and may cause fainting. 
Bruising at the blood draw site may occur, but can be prevented or lessened by applying pressure for 
several minutes. Intramuscular injection also causes transient discomfort. Infection at the site of blood 
drawing or vaccination is extremely unlikely as alcohol swabbing and sterile equipment will be used.  
The use of plasmid DNA has the potential to cause an allergic reaction due to the presence of bacterial 
endotoxin. However, each lot of DNA will be tested for endotoxin to ensure that endotoxin content does 
not exceed USP specifications. Antibodies to DNA may potentially develop in DNA vaccine recipients. 
However, the development of such antibodies in response to DNA vaccination is rare and has not been 
associated with disease in animals or humans to date. 
7.2.2 
Potential toxicity related to the TDS-IM device 
The following are anticipated adverse reactions based on the previous clinical studies of TDS-IM based 
DNA vaccine delivery. All known risks and precautions described are explained in detail in the informed 
consent. 
Local Reactions 
Local site reactions associated with the use of the TDS-IM device in humans include acute pain 
associated with the localized muscle contractions during the application of electroporation in virtually all 
subjects. Mild, transient bleeding at the sites of electrode/needle penetration is commonly observed 
following removal of the device. Soreness, erythema, and/or induration of mild to moderate severity are 
commonly reported at the administration site. In one instance, local site soreness transiently graded as 
severe was reported in association with a device technical error related to needle deployment wherein the 
procedure was suspended prior to the application of electroporation. Other injection site findings of mild 
severity including mild bruising, hematoma, and paresthenia have been reported occasionally. All 
injection site reactions have typically resolved within 24–72 hours following administration, but, in rare 
instances, mild local tenderness has been reported to persist for up to one week. 
For the TDS-IM device there is a theoretical risk that excessive energy could be delivered to the local 
tissues of the subject, resulting in more pronounced local reactions than have been observed to date. 
However, the TDS Pulse Stimulator incorporates multiple redundant mitigations to prevent this hazard, 
including performance of a pre-pulse safety check and the use of multiple circuits that monitor total 
energy delivered and will which will terminate energy delivery. There have been no reported occurrences 
of excessive energy delivery in any of the nonclinical and clinical studies conducted with the pulse 
stimulator to date. 

Washington University School of Medicine 
Protocol Date 08/03/23, A5 
Siteman Cancer Center 
Pancreatic Cancer SPORE 
Neoantigen DNA vaccines  
 
Page 38 of 69 
Systemic Reactions 
Systemic adverse events reported during the studies utilizing the TDS-IM and judged to be possibly 
related to the study product and/or delivery device have been generally mild to moderate in severity and 
include flu-like symptoms, headache, fever, dizziness, malaise, fatigue, arthralgia, myalgia, and aphthous 
stomatitis. Transient elevation in serum creatine phosphokinase of mild to moderate severity and judged 
to be associated with procedure administration have been reported in a small minority of subjects. Two 
subjects have reported the onset of severe fatigue within 24 hours of dosing that was resolved by the 
following day and was not observed following subsequent doses. 
In approximately 1% of procedure administrations, one or more symptoms consistent with a vasovagal 
reaction (i.e., dizziness / lightheadedness, hypotension, diaphoresis, and/or skin pallor) have been 
observed immediately after procedure application. In rare instances, these reactions have progressed to 
brief syncope, particularly in subjects with pre-existing risk factors for syncope (e.g., pronounced sinus 
bradycardia). In all cases, the subjects have rapidly recovered from the syncope without incident.  
A theoretical risk associated with the use of DNA vaccines is the possibility of uptake of the vaccine 
candidate into non-target tissues and/or integration of the vaccine DNA into the genomic DNA of the test 
subject. As described in Section 7.3 below, nonclinical studies have been performed to assess the 
potential for systemic biodistribution and genomic integration following TDS based delivery multiple DNA 
vaccine candidates, including a tyrosinase melanoma DNA vaccine candidate utilizing the same vector 
backbone (pING) as the Mammaglobin-A DNA vaccine candidate. To date, there has been no evidence 
of significant systemic uptake or genomic integration events following TDS based DNA vaccine 
administration.  
 
Toxicity monitoring and management 
Toxicity will be characterized according to the National Cancer Institute Common Terminology Criteria for 
Adverse Events version 4.0 (CTCAE). Subjects who are immunized with the plasmid DNA vaccine will be 
evaluated at the time of each vaccination on weeks 1, 5, 9, 13, 17, and 21. Follow up on subject well-
being will be performed by telephone on the first, second, or third day after each vaccination. All 
information will be recorded on case report forms. Adverse events will be reported to the Quality 
Assurance and Safety Monitoring Committee of the Siteman Cancer Center/Sidney Kimmel 
Comprehensive Cancer Center, the Institutional Review Board, the Institutional Biosafety Committee, the 
Office of Biotechnology Activities and the Food and Drug Administration as detailed in Section 11 
Adverse Event Reporting. 
Significant local inflammation will be treated with cold packs and oral analgesics as indicated. Skin 
ulceration at the vaccine site will be treated with local wound care and antibiotics as indicated. 
Development of signs and/or symptoms of autoimmune involvement will be initially treated conservatively 
with analgesics. More aggressive intervention (systemic corticosteroids) will be used as necessary and 
will result in termination of future vaccine inoculation. 
 
Dose modifications 
No dose modifications are planned. If a subject develops an adverse event that is classified as possibly, 
probably, or definitely associated with protocol therapy, this may result in removal of the subject from 
protocol therapy as outlined in Section 10 Removal of Subjects from Protocol Therapy. Protocol 
Stopping Criteria are outlined in Section 12 Data and Safety Monitoring. 
 

Washington University School of Medicine 
Protocol Date 08/03/23, A5 
Siteman Cancer Center 
Pancreatic Cancer SPORE 
Neoantigen DNA vaccines  
 
Page 39 of 69 
8 
STUDY CALENDAR 
The window for Step 0 procedures is up to 28 days prior to enrollment.  Enrollment must occur within 28 
days of the patient starting on adjuvant chemotherapy.  The window for Step 1 screening procedures is 
14 days prior to first vaccine dose, with the exception of Immune monitoring labs and pathology review, 
which should be performed immediately after informed consent.  Informed consent should be obtained 
within 16 weeks of surgical resection, but there is no window for informed consent prior to study 
enrollment in order to allow time for vaccine creation. 
Skinfold measurements must be performed at site of vaccine injections (deltoids and thighs) during Step 
0 screening, Step 1 screening, and prior to each vaccine dose. 
 
Enrollment 
Eligibility Re-
Check 
Wk 1f 
Wk 5f 
Wk 9f 
Wk 
13f 
Wk 
17f 
Wk 
21f 
Wk 
25f 
Wk 
73h 
Neoantigen DNA 
vaccinea,i 
 
 
X 
X 
X 
X 
X 
X 
 
 
Informed consent 
X 
 
 
 
 
 
 
 
 
 
Demographics 
X 
 
 
 
 
 
 
 
 
 
Medical history 
X 
 
 
 
 
 
 
 
 
 
Physical exam 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
Concurrent Meds 
X ------------------------------------------------------------------------------------------------------------------------ X 
 
Vital signs 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
Height 
X 
 
 
 
 
 
 
 
 
 
Weight 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
Performance Status 
X 
X 
X 
X 
X 
X 
X 
X 
X 
 
CBC w/diff, plts 
X 
X 
 
X 
X 
X 
X 
X 
X 
 
CMPb 
X 
X 
 
X 
X 
X 
X 
X 
X 
 
INR 
X 
X 
 
 
 
 
 
 
 
 
EKG  
X 
X 
 
 
 
 
 
 
 
 
Radiologic 
evaluation 
Radiologic measurements will be performed as clinically indicated. Documentation of tumor recurrence or 
metastatic disease must be provided for patients removed from study for progressive disease. 
Adverse event 
evaluation 
 
 
Xg 
Xg 
Xg 
Xg 
Xg 
Xg 
X 
 
B-HCG 
Xe 
Xe 
Xe 
Xe 
Xe 
Xe 
Xe 
Xe 
 
 
Immune monitoringc 
X 
 
X 
X 
X 
X 
X 
X 
X 
X 
Pathology Reviewd 
X 
 
 
 
 
 
 
 
 
 
a: Personalized polyepitope pancreatic cancer DNA vaccine 
b: Albumin, alkaline phosphatase, total bilirubin, bicarbonate, BUN, calcium, chloride, creatinine, glucose, LDH, phosphorus, 
potassium, total protein, SGOT [AST], SGPT [ALT], sodium. 
c: Immune monitoring labs to consist of 7 10mL green top heparin tubes for isolation of PBMCs. During adjuvant therapy, these 
should occur at a clinic visit associated with the restaging scan, as well as at each follow up visit. This will yield approximately 50-
200 x106 PBMCs per collection. 
d: Pathology review to confirm diagnosis of adenocarcinoma and evaluate tumor cellularity.  
e: Serum or urine pregnancy test (women of childbearing potential). 
f: +/- 7 days with at least 21 days between vaccinations  
g: Adverse event evaluation on day of vaccine and by telephone contact on the first, second, or third day after vaccine. 
h: ± 14 days; follow-up or telephone contact at week 73 and annually thereafter if the patient is alive and available for follow-up 
i: patients may discontinue participation if vaccine production is unable to be completed 

Washington University School of Medicine 
Protocol Date 08/03/23, A5 
Siteman Cancer Center 
Pancreatic Cancer SPORE 
Neoantigen DNA vaccines  
 
Page 40 of 69 
9 
CRITERIA FOR RESPONSE 
 
Primary objective: safety 
Assessment of plasmid DNA safety will include both clinical observation and laboratory evaluation. Safety 
will be closely monitored after injection with eight or more clinical and laboratory assessments in the first 
24 weeks of the trial. The following parameters will be assessed following vaccination: 
(1) 
Local signs and symptoms 
(2)  
Systemic signs and symptoms 
(3) 
Laboratory evaluations, including blood counts and serum chemistries 
(4) 
Adverse, and serious adverse events 
Toxicity will be graded according to the National Cancer Institute Common Terminology Criteria for 
Adverse Events v4.0. 
 
Secondary and exploratory objectives: evaluation of the immune response 
9.2.1 
Introduction to immune monitoring 
The secondary endpoint is to evaluate the immunogenicity of the neoantigen DNA vaccine. 
Immunogenicity will be measured by ELISPOT analysis, a surrogate for CD4 and CD8 T cell function. 
Additionally, an exploratory endpoint is to evaluate the immunogenicity of the neoantigen DNA vaccine as 
measured by multiparametric flow cytometry. In both assays the quantity and quality of antigen-specific T 
cells is determined; the ELISPOT analysis is based on measuring the frequencies of IFN-γ producing T 
cells in response to polyepitope antigen, whereas the multiparametric flow cytometry assesses 
phenotypic as well as functional characteristics of epitope-specific T cells. In the proposed study, blood 
samples will be collected at multiple time points (n=8) and PBMC isolated and cryopreserved. Upon 
completion of the vaccination protocol, all samples will be analyzed simultaneously in order to minimize 
assay-to-assay variation. 
9.2.2 
Sample collection and processing 
During Step 0 screening, tumor samples will be reviewed by a pathologist with expertise in pancreatic 
cancer who will review the operative specimen slides to verify tumor cellularity and quantity sufficient for 
proceeding. Once tissue availability is confirmed at the site, specimens from outside Washington 
University will be shipped to the Gillanders lab at the following address: Gillanders Lab, 425 S. Euclid 
Ave. CSRB 3344, St. Louis, MO 63110.  The corresponding FFPE blocks will be punched with a 
disposable 1mm biopsy punch. 6-8 full thickness punches will be taken from areas of high tumor 
cellularity and divided evenly into two DNA LoBind Eppendorf tubes (one tube for DNA and one for RNA). 
To identify somatic mutations, DNA and RNA will be extracted from the FFPE preserved tissue by the 
Center for Human Immunology and Immunotherapy Programs (CHiiPS) core. All tissue selected for 
sequencing will be processed into a single-cell suspension by mechanical and enzymatic digestion, and 
used to extract nucleic acids. Tumor DNA + RNA will then undergo tumor exome and tumor cDNA-
capture sequencing, respectively at the Genome Institute. Normal genomic DNA will be isolated from 
PBMCs by the CHiiPS core or Gillanders laboratory personnel for normal exome sequencing at The 
Genome Institute.  
Patients will undergo their first blood draw during Step 0 screening to establish a baseline for immune 
monitoring and for creation of the vaccine. This first blood draw will be shipped ambient same day of draw 
for overnight delivery to the Gillanders lab at the following address:  Gillanders Lab, 425 S. Euclid Ave. 
CSRB 3344, St. Louis, MO 63110. Additional optional pre-vaccine blood samples will be obtained midway 
through chemotherapy and at the end of chemotherapy coinciding as closely as possible to the restaging 
scans, and also shipped overnight same day of draw to the Gillanders lab. Immune monitoring labs will be 
draw prior to vaccination on weeks 1, 5, 9, 13, 17, and 21. Post- vaccine immune monitoring labs will be 
drawn on weeks 25, and 73. Seven collection tubes (BD Vacutainer® sodium heparin (green top), REF 
367874, 10 mL each for a total of approximately 60 mL) are filled by venipuncture at each time point. 
Blood samples will be transported to Dr. Gillanders’/JHU equivalent laboratory (6th floor Clinical Sciences 

Washington University School of Medicine 
Protocol Date 08/03/23, A5 
Siteman Cancer Center 
Pancreatic Cancer SPORE 
Neoantigen DNA vaccines  
 
Page 41 of 69 
Research Building, room 6664) within one hour of collection. PBMC will be obtained by Ficoll-Hypaque 
gradient centrifugation and cryopreserved in 10% DMSO according to standard procedures. Blood will be 
batch shipped after Week 9 collection and Week 21 collection to the Gillanders lab at the address listed 
above.  Exome sequencing of PBMC will be performed to obtain germline sequences. 
9.2.3 
ELISPOT Analysis (Secondary Objective) 
ELISPOT analysis, will be performed as previously described [78-82]. PBMC from subjects in the phase 
1b clinical trial will be tested for secretion of IFN-γ by ELISPOT assay. PBMC will be plated at various 
concentrations starting at 300,000 cells per well, in triplicate, following the protocol previously described 
by Dr. Mohanakumar and colleagues. PBMC will be co-cultured with the mutant and mesothelin peptides 
encoded by the polyepitope construct. As negative controls, PBMC will be incubated in medium alone or 
stimulated with matching wild type peptides. As a positive control we will include a mix of viral peptides, 
CEF, containing immunodominant epitopes for multiple common HLA alleles from influenza virus, 
cytomegalovirus, and Epstein-Barr virus. After 24-48 hours, the plates will be developed and the spots 
counted in an ImmunoSpot Series I analyzer (Cellular Technology). 
9.2.4 
Multi-parametric flow cytometry (Exploratory Objective) 
Patient-derived PBMC will be used for functional assays to characterize immunity to personalized 
pancreatic cancer DNA vaccines. Polyfunctional CD8 T cell responses will be determined after stimulation 
of cultured PBMC with polyepitope pulsed autologous PBMC using muti-parameter flow cytometry. 
Fluorescently labeled MHC class I tetramers expressing vaccine-encoded peptides will be used to gate 
on peptide-specific T cells. The choice of tetramers to be used in these analyses will be dictated by the 
neoantigens and mesothelin antigens used for a given patient’s personalized vaccine, and the patient’s 
HLA phenotype. We will also simultaneously stain tetramer-positive and tetramer-negative CD8 T cells for 
a variety of well-accepted markers that report the functional status of antigen specific CD8 T cells. These 
include the cell surface markers PD-1, CTLA-4, LAG-3, and TIM-3 (markers of T cell inactivation), the 
proliferation marker Ki-67, and the cellular activation markers ICOS, granzyme B, TNF-α and IFN-γ. We 
have already validated all of our staining reagents and demonstrated that specific combinations of mAb 
labeled with different fluorescent tags can be used together allowing for multi-color analysis in a single 
run. Analyses will be performed using either a BD Fortessa (6-color analysis) or BD LSR II cytometers 
(11-color analysis). A positive effect of immunotherapy is expected to show increased numbers of 
tetramer-positive cells that also display decreased expression of PD-1, CTLA-4, LAG3, and TIM3, and 
increased staining for Ki-67, ICOS, granzyme B, TNF-β and IFN-γ.  
PBMC aliquots (2x106 cells/mL) will be stimulated with individual peptides encoded by the polyepitope 
vaccine and cultured in the presence of IL-2 (50U/mL). Ten days after activation, cells will be harvested, 
washed and restimulated with irradiated (10,000 rads) peptide-pulsed autologous PBMC for 16h (in the 
presence of Brefeldin A) and stained with antibodies for the various markers listed above. Cells will be 
analyzed by the Immune Monitoring Core, also known as the Center for Human Immunology and 
Immunotherapy Programs (CHiiPs), headed by Dr. Robert Schreiber. Cultured cells stimulated with 
unpulsed PBMC will be used as negative control, and cells stimulated with CEF-pulsed PBMC will be 
used as positive controls using an appropriate tetramer. One advantage of using 10 day 
activated/antigen-driven PBMC to measure polyfunctional responses is the expected relative high 
frequency of polyepitope-specific T cells providing a larger sample size for statistically significant data 
analysis. 
 
Additional Exploratory objectives 
We will also monitor the function and phenotype of antigen-specific T cells using time-of-flight mass 
spectrometry (CyTOF). The CyTOF is a next-generation technology based on inductively coupled plasma 
mass spectrometry (ICP-MS) that employs antibodies or peptide-MHC tetramers labeled with heavy metal 
isotopes instead of fluorophores to identify cell associated proteins of interest [83, 84]. This instrument 
permits investigators to probe up to 49 parameters simultaneously on individual cells thus allowing for 
cellular phenotyping at unprecedented depth. In addition, it facilitates the effective analysis of 
lymphocytes whose numbers may be limited such as those derived from human cancer patients. The 
Washington University Center for Human Immunology and Immunotherapy Programs has now obtained 

Washington University School of Medicine 
Protocol Date 08/03/23, A5 
Siteman Cancer Center 
Pancreatic Cancer SPORE 
Neoantigen DNA vaccines  
 
Page 42 of 69 
and installed two CyTOF2 instruments that are being used successfully under the guidance of Dr. 
Stephen Oh, Assistant Professor of Medicine and Director of Mass Cytometry for CHiiPs. 
Clinical responses and time to disease progression will be evaluated with physical examination and 
diagnostic imaging as clinically indicated. 
 
10 
REMOVAL OF SUBJECTS FROM PROTOCOL THERAPY 
 Removal of subjects from protocol therapy 
Subjects may be removed from protocol therapy if any one or more of the following events occur: 
(1) 
Development of recurrent or metastatic disease prior to the initial vaccination will result in 
stopping vaccine development at its current stage; 
(2) 
Development of recurrent or metastatic disease during the vaccination period will result in 
stopping further vaccinations 
(3) 
Intercurrent illness that prevents further administration of protocol therapy; 
(4) 
Pregnancy; 
(5) 
Type 1 hypersensitivity reaction associated with protocol therapy; 
(6) 
Grade 2 systemic or injection site adverse event classified as possibly, probably, or definitely 
associated with protocol therapy that does not resolve to at least grade 1 prior to the next 
scheduled treatment; 
(7) 
Grade 3 or 4 systemic or injection site adverse event classified as possibly, probably, or definitely 
associated with protocol therapy; 
(8) 
Any significant autoimmune disease or phenomena presumed to be related to protocol therapy; 
(9) 
Unable to complete vaccine production; 
(10) 
Subject refusal to continue protocol therapy and/or observations; 
(11) 
Significant protocol violation or noncompliance, either on the part of the subject or investigator(s); 
(12) 
The principal investigator or study sponsor believes it is in the subject's best interest to 
discontinue participation in the study; 
(13) 
Administrative reasons, e.g., study termination by the principal investigator, Siteman Cancer 
Center/Sidney Kimmel Comprehensive Cancer Center, HRPO, FDA, or other group. 
Please note that even if a subject is removed from protocol therapy, they will continue to be followed for 
adverse events. 
 Voluntary subject withdrawal 
The subject has the right to voluntarily withdraw from the study at any time for any reason without 
prejudice to her future medical care by the physician or at the institution. 
For any subject who withdraws consent, the date and reason for consent withdrawal should be 
documented. Subject data will be included in the analysis up to the date of the consent withdrawal. 
 Procedure for discontinuation 
The procedure to be followed at the time a subject either discontinues participation or is removed from the 
study is: 
(1) 
Check for the development of adverse events. 
(2) 
Complete the End-of-Study form and include an explanation of why the subject is withdrawing or 
withdrawn. 

Washington University School of Medicine 
Protocol Date 08/03/23, A5 
Siteman Cancer Center 
Pancreatic Cancer SPORE 
Neoantigen DNA vaccines  
 
Page 43 of 69 
(3) 
Attempt to perform follow-up evaluations as outlined above. 

Washington University School of Medicine 
Protocol Date 08/03/23, A5 
Siteman Cancer Center 
Pancreatic Cancer SPORE 
Neoantigen DNA vaccines  
 
Page 44 of 69 
11 
ADVERSE EVENT REPORTING 
The entities providing oversight of safety and compliance with the protocol require reporting as outlined 
below.  Please refer to Appendix A for definitions and Appendix B for a grid of reporting timelines. 
Adverse events will be tracked from start of treatment through 30 days following the last day 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 
Refer to the data submission schedule in Section 14 for instructions on the collection of AEs in the EDC. 
Reporting requirements for Washington University study team may be found in section 11.1.  Reporting 
requirements for secondary site study teams participating in Washington University-coordinated research 
may be found in Section 1.2. 
 
 Sponsor-Investigator Reporting Requirements 
11.1.1 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. 
11.1.2 Reporting to the Quality Assurance and Safety Monitoring Committee 
(QASMC) at Washington University 
The Washington University Sponsor Investigator (or designee) is required to notify the 
QASMC of any unanticipated problems involving risks to participants or others occurring 
at WU or BJH or SLCH institution that have 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 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. 
 
For events that occur at secondary sites, the Washington University Sponsor Investigator 
(or designee) 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. 
11.1.3 Reporting to Ichor Medical Systems 
Since the TDS-IM device may be involved with the occurrence of adverse events, Ichor 
Medical Systems will be included in the Adverse Event reporting plan. Reports to Ichor 
for SAEs or for adverse events with possible relationship to the device should follow the 
timing for reporting to the WUSM Human Research Protection Office and should be 
directed to Drew Hannaman at dhannaman@ichorms.com.  The Washington University 
PI will be responsible for submitting all MedWatch forms from secondary sites to Ichor 
Medical Systems. 
 

Washington University School of Medicine 
Protocol Date 08/03/23, A5 
Siteman Cancer Center 
Pancreatic Cancer SPORE 
Neoantigen DNA vaccines  
 
Page 45 of 69 
11.1.4 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 A 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 A) no later than 15 calendar days after it is 
determined that the information qualifies for reporting.  Report an adverse event 
(refer to Appendix A) 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 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”). 
11.1.5 Reporting to the Institutional Biosafety Committee 
In accordance with institutional policies and NIH guidelines, any unanticipated problems 
must be reported to the Institutional Biological and Chemical Safety Committee (IBC) at 
Washington University School of Medicine. 
 

Washington University School of Medicine 
Protocol Date 08/03/23, A5 
Siteman Cancer Center 
Pancreatic Cancer SPORE 
Neoantigen DNA vaccines  
 
Page 46 of 69 
The Washington University Sponsor-Investigator (or designee) must report the following 
events to the Biosafety Officer at the time of submission to HRPO: 
• 
Any overt personnel exposure to recombinant DNA-containing material, whether 
or not that exposure leads to illness 
• 
Any significant spill of recombinant DNA-containing material outside of a biological 
safety cabinet, where a significant spill is: 
o 
A spill of recombinant risk group 1 agent-containing material which 
requires remediation by EH&S or other first responders 
o 
A spill of recombinant risk group 2 agent-containing material which is 
greater than 1 liter 
o 
Any size spill of risk group 3 agent-containing material or material the IBC 
has mandated to be handled using BSL2+ practices and procedures 
o 
Any incident which results in the release of recombinant DNA to the 
environment 
 
In addition, the IBC must be informed of the following events: 
• 
Any serious adverse event which is both unexpected and associated with the use 
of the gene transfer product 
 
The Biosafety Officer may be contacted at ehsibc@wustl.edu. 
 
When submitting reports to the IBC, the FDA MedWatch form will be used.  The 
Washington University Sponsor-Investigator (or designee) will be responsible for 
submitting all MedWatch forms from secondary sites to the IBC.   
 
WUSM IBC guidelines specify that any study modifications related to the investigational 
agent as well as the IRB renewal paperwork should be sent to the IBC for approval (if a 
modification) or acknowledgment (if an annual renewal).  The IRB and IBC will review all 
submissions simultaneously. 
11.1.6 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. 
 
 
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 A, 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 FDA Form 3500a (MedWatch) and Washington University’s cover sheet 
(Appendix C).  A formal written report must be sent to the Washington University Sponsor-
Investigator and designee within 4 calendar days (for fatal or life-threatening suspected adverse 
reactions) or 11 calendar days (for serious unexpected suspected 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, IBC, and 
Ichor as needed. 
 

Washington University School of Medicine 
Protocol Date 08/03/23, A5 
Siteman Cancer Center 
Pancreatic Cancer SPORE 
Neoantigen DNA vaccines  
 
Page 47 of 69 
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.  
Washington University IRB approval is not required for protocol exceptions occurring at secondary 
sites. 
 
 
Exceptions to Expedited Reporting 
Events that do not require expedited reporting as described in Section 11.1 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. 
 
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 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 
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 6 months beginning after study activation at Washington University, 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 and accrual by site 
• 
Objectives of protocol with supporting data and list the number of participants who have met 
each objective 

Washington University School of Medicine 
Protocol Date 08/03/23, A5 
Siteman Cancer Center 
Pancreatic Cancer SPORE 
Neoantigen DNA vaccines  
 
Page 48 of 69 
• 
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 
• 
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 11). 
Refer to the Washington University Quality Assurance and Safety Monitoring Committee Policies and 
Procedures for full details on the responsibilities of the DSMC.  This is located on the QASMC website at 
https://siteman.wustl.edu/research/clinical-research-resources/protocol-office-prmcqasmc/.   
 NIH Recombinant DNA Advisory Committee 
12.1.1 Initiation of the clinical investigation 
Appendix M-I-C-1 of the NIH Guidelines for Research Involving Recombinant DNA Molecules specify: 
No later than 20 working days after enrollment (see definition of enrollment in Section I-E-7) of the 
first research participant in a human gene transfer experiment, the Principal Investigator(s) shall 
submit the following documentation to NIH OBA: (1) a copy of the informed consent document 
approved by the Institutional Review Board (IRB); (2) a copy of the protocol approved by the 
Institutional Biosafety Committee (IBC) and IRB; (3) a copy of the final IBC approval from the clinical 
trial site; (4) a copy of the final IRB approval; (5) a brief written report that includes the following 
information: (a) how the investigator(s) responded to each of the RAC’s recommendations on the 
protocol (if applicable); and (b) any modifications to the protocol as required by FDA; (6) applicable 
NIH grant number(s); (7) the FDA Investigational New Drug Application (IND) number; and (8) the 
date of the initiation of the trial. The purpose of requesting the FDA IND number is for facilitating 
interagency collaboration in the Federal oversight of human gene transfer research. 
12.1.2 Annual reports 
Appendix M-I-C-3 of the NIH Guidelines for Research Involving Recombinant DNA Molecules specify: 
Within 60 days after the one-year anniversary of the date on which the investigational new drug (IND) 
application went into effect, and after each subsequent anniversary until the trial is completed, the 
Principal Investigator (or delegate) shall submit the information set forth in (a), (b), and (c). When 
multiple studies are conducted under the single IND, the Principal Investigator (or delegate) may 
choose to submit a single annual report covering all studies, provided that each study is identified by 
its OBA protocol number. 
(a) Clinical Trial Information. A brief summary of the status of each trial in progress and each trial 
completed during the previous year. The summary is required to include the following information for 
each trial: (1) the title and purpose of the trial; (2) clinical site; (3) the Principal Investigator; (4) clinical 
protocol identifiers, including the NIH OBA protocol number, NIH grant number(s) (if applicable), and 
the FDA IND application number; (5) participant population (such as disease indication and general 
age group, e.g., adult or pediatric); (6) the total number of participants planned for inclusion in the 
trial; the number entered into the trial to date; the number whose participation in the trial was 
completed; and the number who dropped out of the trial with a brief description of the reasons; (7) the 
status of the trial, e.g., open to accrual of subjects, closed but data collection ongoing, or fully 
completed, and (8) if the trial has been completed, a brief description of any study results. 
(b) Progress Report and Data Analysis. Information obtained during the previous year's clinical and 
non-clinical investigations, including: (1) a narrative or tabular summary showing the most frequent 
and most serious adverse experiences by body system; (2) a summary of all serious adverse events 
submitted during the past year; (3) a summary of serious adverse events that were expected or 

Washington University School of Medicine 
Protocol Date 08/03/23, A5 
Siteman Cancer Center 
Pancreatic Cancer SPORE 
Neoantigen DNA vaccines  
 
Page 49 of 69 
considered to have causes not associated with the use of the gene transfer product such as disease 
progression or concurrent medications; (4) if any deaths have occurred, the number of participants 
who died during participation in the investigation and causes of death; and (5) a brief description of 
any information obtained that is pertinent to an understanding of the gene transfer product’s actions, 
including, for example, information about dose-response, information from controlled trials, and 
information about bioavailability. 
(c) A copy of the updated clinical protocol including a technical and non-technical abstract. 
 
13 
STATISTICAL CONSIDERATIONS 
 Experimental Design 
This is a phase 1, open-label, trial of a neoantigen DNA vaccine strategy. Twenty patients who have had 
an R0/R1 surgical resection, and who have completed adjuvant chemotherapy will be enrolled. At our 
institution, pancreatic cancer patients undergo routine staging during (three months) and after adjuvant 
chemotherapy. If there is no evidence of disease, patients will be eligible for enrollment. After enrollment, 
exome sequencing of tumor/normal DNA and cDNA-capture sequencing of the tumor will be performed to 
identify somatic mutations present in the tumor and confirm expression of these mutations at the mRNA 
level. Epitope prediction algorithms will then be used to prioritize neoantigens for inclusion in neoantigen 
DNA vaccines, and the neoantigen DNA vaccines will be designed and manufactured. The neoantigen 
DNA vaccines will be administered following completion of adjuvant chemotherapy. The neoantigen DNA 
vaccines will be administered at six time points, weeks 1, 5, 9, 13, 17, and 21.  
13.1.1 Sample size and accrual 
Development of cancer vaccines is currently an area of intense research interest. The traditional 
paradigm for phase 1 clinical trials has been heavily influenced by phase 1 clinical trials of 
chemotherapeutic agents, where dose escalation designs are appropriate given the rather narrow dose 
versus safety concerns of these agents. However, there are major differences between these two classes 
of therapeutics that have important implications for early clinical development. Specifically, the phase 1 
concept of dose escalation to find a maximum-tolerated dose does not apply to most therapeutic cancer 
vaccines. Most therapeutic cancer vaccines are associated with minimal toxicity at a range that is feasible 
to manufacture or administer, and there is little reason to believe that the maximum-tolerated dose is the 
most effective dose [52]. Consistent with recent recommendations published in the statistical literature, 
the general philosophy of this phase I clinical trial is to facilitate a prompt preliminary evaluation of the 
safety and immunogenicity of a personalized pancreatic cancer DNA vaccine strategy by testing a fixed 
dose of vaccine, instead of performing a dose escalation [52]. The sample size (n=15) is chosen to 
provide a reasonable ability to detect serious adverse events associated with vaccine administration. 
Based on extensive simulations regarding the sample size for pilot or translational studies, Piantadosi [83] 
recommended that a sample size of 10 to 20 patients would provide a reasonable precision for estimating 
preliminary information. We therefor expect that, as detailed below, the proposed sample size for this 
study will be adequate in estimating both safety and preliminary immune efficacy data.  The sample size 
of n=15 will consist of EVALUABLE patients.  Patients who screen fail due to sequencing issues such as 
poor sample quality will be replaced. 
13.1.2 Sample Size Calculations for Safety 
The primary objective of this study is to evaluate the safety of the neoantigen DNA vaccines. We have 
chosen to power the study to provide a reasonable ability to detect serious adverse events associated 
with vaccine administration. 
Sample size calculations for safety are expressed in terms of the ability to detect serious adverse events. 
The ability of the study to identify serious adverse events is best expressed by the maximum true rate of 
events that would be unlikely to be observed and the minimum true rate of events that would very likely 
be observed. Table below shows the probabilities of observing 0, or 2 or more serious adverse events 
given variety of hypothesized “true” AE rates. If the true rate is at least 15%, for example, there is >90% 
chance of observing at least 1 serious adverse event among a sample size of n=15 evaluable patients. 

Washington University School of Medicine 
Protocol Date 08/03/23, A5 
Siteman Cancer Center 
Pancreatic Cancer SPORE 
Neoantigen DNA vaccines  
 
Page 50 of 69 
Conversely, there is only 10% chance that we would observe 2 or more serious adverse event among 
n=15 evaluable patients if the true rate is less than 3.5%. Although we believe that the true event rate is 
likely to be quite low, this table presents a range of event rates in an attempt to illustrate the sensitivity of 
this study to identify potential safety problems with the neoantigen DNA vaccines. 
 
Probability of detecting SAE for different safety scenarios 
True Event rate 
Pr(0/15) 
Pr(2+/15) 
0.010 
0.860 
0.010 
0.035 
0.586 
0.100 
0.050 
0.463 
0.171 
0.100 
0.206 
0.451 
0.150 
0.087 
0.681 
13.1.3 Sample Size Calculations for Immunogenicity 
The secondary objective and primary scientific endpoint is to assess the prevalence of antigen-specific T 
cells in the peripheral blood of patients pre- and post-vaccination as measured by ELISPOT.. A power 
analysis was performed using paired t-test for over time differences (e.g., changes in ELISPOT). 
Assuming a moderate correlation between measures taken from the same individual, the designed 
sample size (n=15 evaluable patients) allows us 80% power at 1-sided 0.05 alpha level to detect a 
minimum increase of 0.68*SD, where SD represents the standard deviation of measures for 
immunogenicity. Our preliminary data showed that antigen-specific T cells can be measured with a 
coefficient of variability (CV=SD/Mean) of approximately 20%. Therefore, if assuming similar variability in 
the proposed study, the designed sample size provides 80% power to detect a minimal of 15% change in 
the average antigen-specific T cells. We expect that more power could be achieved because multiple 
measurements will be taken from the same patient and this allows borrowing information across different 
time points. 
 Data Analysis 
As a phase I trial to evaluate the preliminary data on the safety and immunogenicity of a neoantigen 
DNA vaccine strategy, the data analysis for this study is descriptive in nature. Demographic and clinical 
characteristics of the sample, toxicity by grade, as well as response and time to toxicity will be listed and 
summarized using descriptive statistics. 
13.2.1 Primary objective: safety analysis 
Toxicity evaluation is the primary objective of this trial. The data will be descriptive, and standard toxicity 
definitions and criteria will be used as outlined in the National Cancer Institute Common Terminology 
Criteria for Adverse Events version 4.0. 
Since enrollment will be concurrent with the first dose of the personalized pancreatic cancer DNA 
vaccines, all subjects will have received at least one vaccination, and all subjects will provide at least 
some safety data. 
The number and percentage of subjects experiencing each type of adverse event will be tabulated by 
severity, and relationship to treatment. If appropriate, confidence intervals will be used to characterize the 
precision of the estimate. A complete listing of adverse events will also be tabulated, and will provide 
details including severity, relationship to treatment, onset, duration, and outcome. 
Laboratory data measured on a continuous scale will be characterized by summary statistics (mean and 
standard deviation). Boxplots of laboratory data will be generated for baseline values and for values 

Washington University School of Medicine 
Protocol Date 08/03/23, A5 
Siteman Cancer Center 
Pancreatic Cancer SPORE 
Neoantigen DNA vaccines  
 
Page 51 of 69 
measured during and after protocol therapy at each specific time point. Each boxplot will show the 
median, 1st and 3rd quartiles. Outliers will be individually plotted in a separate graph, as appropriate. 
13.2.2 Secondary and Exploratory objectives: immune response as measured by ELISPOT 
analysis and multiparametric flow cytometry 
Immune response as measured by ELISPOT analysis is the secondary objective of this trial. Immune 
response as measured by multiparametic flow cytometry is an exploratory objective of this trial. The 
frequency of antigen-specific T cells at each time will be summarized using means, standard deviations 
and medians, and the change over time will also be compared using two-ANOVA for repeated 
measurement data or Friedman rank-sum test as appropriate. The immunogenicity of the neoantigen 
DNA vaccine will also be analyzed qualitatively by summarizing the phenotypic and functional 
characteristics of antigen-specific T cells. Responses will be considered positive if the number of T cells 
after vaccination is greater than two standard deviations above the mean before vaccination [82]. The 
frequency of positive responses at each time point will be assessed and binomial response rates with 
95% confidence interval estimates will be presented. In addition to presenting the binomial response 
rates, graphical and tabular summaries of the underlying distributions will be made. 
13.2.3 Additional Exploratory objectives 
Blood samples will be obtained at multiple time points (prior to vaccination, during, and post-vaccination) 
in order to procure PBMC for correlative studies. Functional studies will include ELISPOT, T cell poly-
functionality by intra-cellular cytokine and degranulation analysis using multi-parametric flow cytometry, 
as detailed in Section 9. 
Time to disease progression will be evaluated with physical examination and diagnostic imaging as 
clinically indicated. The time to disease progression will be described using Kaplan-Meier product limited 
method. The median progression-free survival (PFS), median overall survival (OS), and their 95% CIs will 
be estimated. The association between immunogenicity and PFS or OS will be explored by comparing the 
survival curves between immune responders versus non-responders. To determine whether the observed 
difference is larger than might be expected by chance, a permutation test will be used to compare the 
observed test statistic to the distribution of test statistics that would be seen if there were no difference 
between the two studies. Specifically, we will randomly shuffle the status of response and calculate the 
test statistic from the shuffled data. This procedure will be repeated 10,000 times and the resultant testing 
statistics will provide an accurate representation of the null distribution. The observed test statistics of 
between-study differences will be compared to the null distributions. For each outcome, the permuted p-
value will be the fraction of permuted samples that resulted in a small statistic than the original sample 
[85]. 

Washington University School of Medicine 
Protocol Date 08/03/23, A5 
Siteman Cancer Center 
Pancreatic Cancer SPORE 
Neoantigen DNA vaccines  
 
Page 52 of 69 
14 
DATA MANAGEMENT 
Data collected will be collected using paper case report forms or OnCore electronic data capture forms. 
Forms must be completed within 28 days of time point. 
Case Report Form 
Submission Schedule 
Original Consent Form 
Prior to registration 
Registration Form 
Eligibility Form 
Demographics Form 
On-Study Form 
Medical History Form 
Prior to starting treatment 
Physical Exam Form 
Enrollment 
Mid chemo 
End of chemo 
Week 1 
Week 5 
Week 9 
Week 13 
Week 17 
Week 21 
Week 25 
Week 73 
Vaccine Administration Form 
Week 1 
Week 5 
Week 9 
Week 13 
Week 17 
Week 21 
Immune Monitoring Form 
Enrollment 
Mid chemo 
End of chemo 
Week 1 
Week 5 
Week 9 
Week 13 
Week 17 
Week 21 
Week 25 
Week 73 
Imaging Form 
Mid chemo 
End of chemo 
Toxicity Form 
Week 1 
Week 5 
Week 9 
Week 13 
Week 17 
Week 21 
Week 25 
Concomitant Medications Form 
Continuous through Week 25 
Follow Up Form 
Week 25 
Week 73 
Year 2 and annually thereafter  
MedWatch Form 
See Section 11.0 for reporting requirements 
 

Washington University School of Medicine 
Protocol Date 08/03/23, A5 
Siteman Cancer Center 
Pancreatic Cancer SPORE 
Neoantigen DNA vaccines  
 
Page 53 of 69 
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. 
 
Adverse Event Collection in the Case Report Forms 
All adverse events that occur beginning with start of treatment (minus exceptions defined in Section 
11.0) 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. 
 
15 
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 
Additional details regarding the auditing policies and procedures can be found at 
https://siteman.wustl.edu/wp-content/uploads/2015/10/QASMC-Policies-and-Procedures-03.31.2015.pdf 
 
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. 

Washington University School of Medicine 
Protocol Date 08/03/23, A5 
Siteman Cancer Center 
Pancreatic Cancer SPORE 
Neoantigen DNA vaccines  
 
Page 54 of 69 
• 
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 DSMC (if applicable), and the Washington University study 
team. 
• 
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, 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. 
 
17 
REGULATORY AND ETHICAL OBLIGATIONS 
 Informed consent 
In accordance with US FDA regulations (21 CFR 50) and guidelines (Federal Register, May 9, 1997, Vol. 
62, Number 90 - ICH Good Clinical Practice Consolidated Guideline) it is the investigator’s responsibility 
to ensure that witnessed informed consent is obtained from the subject before participating in an 
investigational study, after an adequate explanation of the purpose, methods, risks, potential benefits and 
subject responsibilities of the study. Procedures that are to be performed as part of the practice of 
medicine and which would be done whether or not study entry was contemplated, such as for diagnosis 
or treatment of a disease or medical condition, may be performed and the results subsequently used for 
determining study eligibility without first obtaining consent. On the other hand, informed consent must be 
obtained prior to initiation of any screening procedures that are performed solely for the purpose of 
determining eligibility for research. 
Each subject must be given a copy of the informed consent. The original signed consent must be retained 
in the institution’s records and is subject to review by the sponsor, the HRPO and any other applicable 
regulatory agencies responsible for the conduct of the institution. All elements listed in the ICH Good 
Clinical Practice guidelines must be included in the informed consent. 
Informed consent will be obtained by either the principal investigator or by individuals approved by the 
principal investigator and whose names have been submitted to the IRB. Informed consent will be 
obtained from the subject after the details of the protocol have been reviewed. The individual responsible 
for obtaining consent will assure, prior to signing of the informed consent, that the subject has had all 
questions regarding therapy and the protocol answered. 
 Institutional Review Board 
In accordance with US FDA regulations (21 CFR 56) and guidelines (Federal Register, May 9, 1997 Vol. 
62 Number 90 - ICH Good Clinical Practice Consolidated Guideline) all research involving human 
subjects must be reviewed and approved by the local IRB. All modifications to the protocol, consent 
forms, or other study documents must be reviewed and approved by the local IRB. At Washington 
University School of Medicine, the Human Research Protection Office serves as the local IRB. 

Washington University School of Medicine 
Protocol Date 08/03/23, A5 
Siteman Cancer Center 
Pancreatic Cancer SPORE 
Neoantigen DNA vaccines  
 
Page 55 of 69 
 Subject confidentiality 
In order to ensure subject confidentiality, each subject will be assigned a study number. Subject samples 
and medical information will be de-identified and labeled with the study number. The link between subject 
identification and study number will be safeguarded in a secure file in a locked room, and access will be 
restricted to the principal investigator, study coordinator, and other co-investigators as necessary. 
Collected data will be recorded on case report forms. Case report forms will be safeguarded in a locked 
cabinet and/or a password-protected secure computer drive and access will be restricted to the principal 
investigator, study coordinator, and other co-investigators as necessary. Subject medical information 
related to, or obtained for the purposes of this trial are confidential, and disclosure to third parties is 
prohibited. The exception is regulatory authorities including the FDA, NIH/OBA, and the local IRB. Data 
from this study must be available for inspection on request of regulatory authorities including the FDA and 
the local IRB. 
 
18 
ADMINISTRATIVE AND LEGAL OBLIGATIONS 
 Study documentation and retention of records 
18.1.1 Study documentation 
Source documents are original documents, data, and records from which the subject’s data are obtained. 
These include but are not limited to hospital records, clinical and office charts, laboratory and pharmacy 
records, diaries, diagnostic imaging studies, and correspondence. 
The principal investigator and staff are responsible for maintaining a comprehensive file of all study-
related documents, suitable for inspection at any time by representatives from the PRMC, HRPO, FDA, 
and any other applicable regulatory agency. 
Pertinent documents in the study file include: 
(1) 
The original protocol with all amendments 
(2) 
Curriculum vitae of principal investigator and co-investigators 
(3) 
Approval notification and any other correspondence with the PRMC, HRPO, NIH RAC and FDA 
Pertinent documents in each individual subject file include: 
(1) 
Informed consent forms 
(2) 
Case report forms 
(3) 
Supporting copies of source documentation 
All original source documentation must be readily available. 
18.1.2 Retention of records 
The principal investigator must retain records related to this study including protocols; amendments; 
IRB/IBC approvals; FDA IND records and other correspondence; completed, signed and dated consent 
forms; patient medical records; case report forms; drug accountability records and any other 
correspondence related to the conduct of the study. 
U.S. FDA regulations (21 CFR 312.62[c]) require that all records pertaining to the conduct of this study, 
must be retained by the responsible investigator for a minimum of 2 years after marketing application 
approval. If no application is filed, these records must be kept 3 years after the investigation is 
discontinued and the U.S. FDA and the applicable local health authorities are notified. 
 Policy regarding research-related injuries 
Washington University School of Medicine investigators and their staffs will try to reduce, control, and 
treat any complications from this research. 

Washington University School of Medicine 
Protocol Date 08/03/23, A5 
Siteman Cancer Center 
Pancreatic Cancer SPORE 
Neoantigen DNA vaccines  
 
Page 56 of 69 
Any subjects who believe that they have been injured as a result of participation in this study will be 
instructed to contact the principal investigator, William E. Gillanders, M.D. at (314) 747-0072. 
Alternatively, they can contact Dr. Jonathan Green, Chairman of the Human Research Protection Office, 
at (800) 438-0445. 
Decisions about payment for medical treatment for research-related injuries will be made by Washington 
University School of Medicine. 
In general, Washington University School of Medicine will provide no long-term medical care or financial 
compensation for research-related injuries. 
 Study termination 
The principal investigator and the Siteman Cancer Center reserve the right to terminate the study. The 
principal investigator will notify the PRMC and HRPO in writing of the study's completion or early 
termination. 

Washington University School of Medicine 
Protocol Date 08/03/23, A5 
Siteman Cancer Center 
Pancreatic Cancer SPORE 
Neoantigen DNA vaccines  
 
Page 57 of 69 
19 
REFERENCES 
1. 
Siegel, R., et al., Cancer statistics, 2014. CA Cancer J Clin, 2014. 64(1): p. 9-29. 
2. 
Rahib, L., et al., Projecting cancer incidence and deaths to 2030: the unexpected burden of 
thyroid, liver, and pancreas cancers in the United States. Cancer Res, 2014. 74(11): p. 2913-21. 
3. 
Conroy, T., et al., FOLFIRINOX versus gemcitabine for metastatic pancreatic cancer. N Engl J 
Med, 2011. 364(19): p. 1817-25. 
4. 
Von Hoff, D.D., et al., Increased survival in pancreatic cancer with nab-paclitaxel plus 
gemcitabine. N Engl J Med, 2013. 369(18): p. 1691-703. 
5. 
Jones, S., et al., Core signaling pathways in human pancreatic cancers revealed by global 
genomic analyses. Science, 2008. 321(5897): p. 1801-6. 
6. 
Clark, C.E., G.L. Beatty, and R.H. Vonderheide, Immunosurveillance of pancreatic 
adenocarcinoma: insights from genetically engineered mouse models of cancer. Cancer Lett, 
2009. 279(1): p. 1-7. 
7. 
Clark, C.E., et al., Dynamics of the immune reaction to pancreatic cancer from inception to 
invasion. Cancer Res, 2007. 67(19): p. 9518-27. 
8. 
Mahadevan, D. and D.D. Von Hoff, Tumor-stroma interactions in pancreatic ductal 
adenocarcinoma. Mol Cancer Ther, 2007. 6(4): p. 1186-97. 
9. 
Porembka, M.R., et al., Pancreatic adenocarcinoma induces bone marrow mobilization of 
myeloid-derived suppressor cells which promote primary tumor growth. Cancer Immunol 
Immunother, 2012. 61(9): p. 1373-85. 
10. 
Sanford, D.E., et al., Inflammatory monocyte mobilization decreases patient survival in pancreatic 
cancer: a role for targeting the CCL2/CCR2 axis. Clin Cancer Res, 2013. 19(13): p. 3404-15. 
11. 
van der Bruggen, P., et al., A gene encoding an antigen recognized by cytolytic T lymphocytes on 
a human melanoma. Science, 1991. 254(5038): p. 1643-7. 
12. 
Peshwa, M.V., et al., Induction of prostate tumor-specific CD8+ cytotoxic T-lymphocytes in vitro 
using antigen-presenting cells pulsed with prostatic acid phosphatase peptide. Prostate, 1998. 
36(2): p. 129-38. 
13. 
Peoples, G.E., et al., Breast and ovarian cancer-specific cytotoxic T lymphocytes recognize the 
same HER2/neu-derived peptide. Proc Natl Acad Sci U S A, 1995. 92(2): p. 432-6. 
14. 
Johnston, F.M., et al., Circulating mesothelin protein and cellular antimesothelin immunity in 
patients with pancreatic cancer. Clin Cancer Res, 2009. 15(21): p. 6511-8. 
15. 
Le, D.T., et al., Safety and survival with GVAX pancreas prime and Listeria Monocytogenes-
expressing mesothelin (CRS-207) boost vaccines for metastatic pancreatic cancer. J Clin Oncol, 
2015. 33(12): p. 1325-33. 
16. 
Lutz, E., et al., A lethally irradiated allogeneic granulocyte-macrophage colony stimulating factor-
secreting tumor vaccine for pancreatic adenocarcinoma. A Phase II trial of safety, efficacy, and 
immune activation. Ann Surg, 2011. 253(2): p. 328-35. 
17. 
Thomas, A.M., et al., Mesothelin-specific CD8(+) T cell responses provide evidence of in vivo 
cross-priming by antigen-presenting cells in vaccinated pancreatic cancer patients. J Exp Med, 
2004. 200(3): p. 297-306. 
18. 
Parmiani, G., et al., Unique human tumor antigens: immunobiology and use in clinical trials. J 
Immunol, 2007. 178(4): p. 1975-9. 
19. 
Sensi, M. and A. Anichini, Unique tumor antigens: evidence for immune control of genome 
integrity and immunogenic targets for T cell-mediated patient-specific immunotherapy. Clin 
Cancer Res, 2006. 12(17): p. 5023-32. 
20. 
Wolfel, T., et al., A p16INK4a-insensitive CDK4 mutant targeted by cytolytic T lymphocytes in a 
human melanoma. Science, 1995. 269(5228): p. 1281-4. 
21. 
Wang, R.F., et al., Cloning genes encoding MHC class II-restricted antigens: mutated CDC27 as 
a tumor antigen. Science, 1999. 284(5418): p. 1351-4. 
22. 
Echchakir, H., et al., A point mutation in the alpha-actinin-4 gene generates an antigenic peptide 
recognized by autologous cytolytic T lymphocytes on a human lung carcinoma. Cancer Res, 
2001. 61(10): p. 4078-83. 
23. 
Brandle, D., et al., A mutated HLA-A2 molecule recognized by autologous cytotoxic T 
lymphocytes on a human renal cell carcinoma. J Exp Med, 1996. 183(6): p. 2501-8. 

Washington University School of Medicine 
Protocol Date 08/03/23, A5 
Siteman Cancer Center 
Pancreatic Cancer SPORE 
Neoantigen DNA vaccines  
 
Page 58 of 69 
24. 
Matsushita, H., et al., Cancer exome analysis reveals a T-cell-dependent mechanism of cancer 
immunoediting. Nature, 2012. 482(7385): p. 400-4. 
25. 
Gubin, M.M., et al., Checkpoint blockade cancer immunotherapy targets tumour-specific mutant 
antigens. Nature, 2014. 515(7528): p. 577-81. 
26. 
Parker, K.C., M.A. Bednarek, and J.E. Coligan, Scheme for ranking potential HLA-A2 binding 
peptides based on independent binding of individual peptide side-chains. J Immunol, 1994. 
152(1): p. 163-75. 
27. 
Rammensee, H., et al., SYFPEITHI: database for MHC ligands and peptide motifs. 
Immunogenetics, 1999. 50(3-4): p. 213-9. 
28. 
Cox, G.J., T.J. Zamb, and L.A. Babiuk, Bovine herpesvirus 1: immune responses in mice and 
cattle injected with plasmid DNA. J Virol, 1993. 67(9): p. 5664-7. 
29. 
Davis, H.L., M.L. Michel, and R.G. Whalen, DNA-based immunization induces continuous 
secretion of hepatitis B surface antigen and high levels of circulating antibody. Hum Mol Genet, 
1993. 2(11): p. 1847-51. 
30. 
Fynan, E.F., et al., DNA vaccines: protective immunizations by parenteral, mucosal, and gene-
gun inoculations. Proc Natl Acad Sci U S A, 1993. 90(24): p. 11478-82. 
31. 
Tang, D.C., M. DeVit, and S.A. Johnston, Genetic immunization is a simple method for eliciting an 
immune response. Nature, 1992. 356(6365): p. 152-4. 
32. 
Ulmer, J.B., et al., Heterologous protection against influenza by injection of DNA encoding a viral 
protein. Science, 1993. 259(5102): p. 1745-9. 
33. 
Wang, B., et al., Gene inoculation generates immune responses against human 
immunodeficiency virus type 1. Proc Natl Acad Sci U S A, 1993. 90(9): p. 4156-60. 
34. 
Donnelly, J.J., et al., DNA vaccines. Annu Rev Immunol, 1997. 15: p. 617-48. 
35. 
Gurunathan, S., D.M. Klinman, and R.A. Seder, DNA vaccines: immunology, application, and 
optimization*. Annu Rev Immunol, 2000. 18: p. 927-74. 
36. 
Stevenson, F.K., et al., DNA fusion gene vaccines against cancer: from the laboratory to the 
clinic. Immunol Rev, 2004. 199: p. 156-80. 
37. 
Dupuis, M., et al., Distribution of DNA vaccines determines their immunogenicity after 
intramuscular injection in mice. J Immunol, 2000. 165(5): p. 2850-8. 
38. 
Aihara, H. and J. Miyazaki, Gene transfer into muscle by electroporation in vivo. Nat Biotechnol, 
1998. 16(9): p. 867-70. 
39. 
Mathiesen, I., Electropermeabilization of skeletal muscle enhances gene transfer in vivo. Gene 
Ther, 1999. 6(4): p. 508-14. 
40. 
Mir, L.M., et al., High-efficiency gene transfer into skeletal muscle mediated by electric pulses. 
Proc Natl Acad Sci U S A, 1999. 96(8): p. 4262-7. 
41. 
Widera, G., et al., Increased DNA vaccine delivery and immunogenicity by electroporation in vivo. 
J Immunol, 2000. 164(9): p. 4635-40. 
42. 
Boyer, J.D., et al., SIV DNA vaccine co-administered with IL-12 expression plasmid enhances 
CD8 SIV cellular immune responses in cynomolgus macaques. J Med Primatol, 2005. 34(5-6): p. 
262-70. 
43. 
Chong, S.Y., et al., Comparative ability of plasmid IL-12 and IL-15 to enhance cellular and 
humoral immune responses elicited by a SIVgag plasmid DNA vaccine and alter disease 
progression following SHIV(89.6P) challenge in rhesus macaques. Vaccine, 2007. 25(26): p. 
4967-82. 
44. 
Hirao, L.A., et al., Combined effects of IL-12 and electroporation enhances the potency of DNA 
vaccination in macaques. Vaccine, 2008. 26(25): p. 3112-20. 
45. 
Hirao, L.A., et al., Intradermal/subcutaneous immunization by electroporation improves plasmid 
vaccine delivery and potency in pigs and rhesus macaques. Vaccine, 2008. 26(3): p. 440-8. 
46. 
Luckay, A., et al., Effect of plasmid DNA vaccine design and in vivo electroporation on the 
resulting vaccine-specific immune responses in rhesus macaques. J Virol, 2007. 81(10): p. 5257-
69. 
47. 
Otten, G., et al., Enhancement of DNA vaccine potency in rhesus macaques by electroporation. 
Vaccine, 2004. 22(19): p. 2489-93. 
48. 
Otten, G.R., et al., Potent immunogenicity of an HIV-1 gag-pol fusion DNA vaccine delivered by in 
vivo electroporation. Vaccine, 2006. 24(21): p. 4503-9. 

Washington University School of Medicine 
Protocol Date 08/03/23, A5 
Siteman Cancer Center 
Pancreatic Cancer SPORE 
Neoantigen DNA vaccines  
 
Page 59 of 69 
49. 
Schadeck, E.B., et al., A dose sparing effect by plasmid encoded IL-12 adjuvant on a SIVgag-
plasmid DNA vaccine in rhesus macaques. Vaccine, 2006. 24(21): p. 4677-87. 
50. 
Rice, J., C.H. Ottensmeier, and F.K. Stevenson, DNA vaccines: precision tools for activating 
effective immunity against cancer. Nat Rev Cancer, 2008. 8(2): p. 108-20. 
51. 
Weiner, D.B., DNA vaccines: Crossing a line in the sand Introduction to special issue. Vaccine, 
2008. 
52. 
Simon, R.M., et al., Clinical trial designs for the early clinical development of therapeutic cancer 
vaccines. J Clin Oncol, 2001. 19(6): p. 1848-54. 
53. 
Bowne, W.B., et al., Coupling and uncoupling of tumor immunity and autoimmunity. J Exp Med, 
1999. 190(11): p. 1717-22. 
54. 
Hawkins, W.G., et al., Immunization with DNA coding for gp100 results in CD4 T-cell independent 
antitumor immunity. Surgery, 2000. 128(2): p. 273-80. 
55. 
Dunn, G.P., et al., A critical function for type I interferons in cancer immunoediting. Nat Immunol, 
2005. 6(7): p. 722-9. 
56. 
Dunn, G.P., C.M. Koebel, and R.D. Schreiber, Interferons, immunity and cancer immunoediting. 
Nat Rev Immunol, 2006. 6(11): p. 836-48. 
57. 
Dunn, G.P., L.J. Old, and R.D. Schreiber, The three Es of cancer immunoediting. Annu Rev 
Immunol, 2004. 22: p. 329-60. 
58. 
Shankaran, V., et al., IFNgamma and lymphocytes prevent primary tumour development and 
shape tumour immunogenicity. Nature, 2001. 410(6832): p. 1107-11. 
59. 
Diamond, M.S., et al., Type I interferon is selectively required by dendritic cells for immune 
rejection of tumors. J Exp Med, 2011. 208(10): p. 1989-2003. 
60. 
Dunn, G.P., et al., Cancer immunoediting: from immunosurveillance to tumor escape. Nat 
Immunol, 2002. 3(11): p. 991-8. 
61. 
Vesely, M.D., et al., Natural innate and adaptive immunity to cancer. Annu Rev Immunol, 2011. 
29: p. 235-71. 
62. 
Vesely, M.D. and R.D. Schreiber, Cancer immunoediting: antigens, mechanisms, and 
implications to cancer immunotherapy. Ann N Y Acad Sci, 2013. 1284: p. 1-5. 
63. 
Klco, J.M., et al., Functional heterogeneity of genetically defined subclones in acute myeloid 
leukemia. Cancer Cell, 2014. 25(3): p. 379-92. 
64. 
Welch, J.S., et al., The origin and evolution of mutations in acute myeloid leukemia. Cell, 2012. 
150(2): p. 264-78. 
65. 
Ding, L., et al., Genome remodelling in a basal-like breast cancer metastasis and xenograft. 
Nature, 2010. 464(7291): p. 999-1005. 
66. 
Ellis, M.J., et al., Whole-genome analysis informs breast cancer response to aromatase inhibition. 
Nature, 2012. 486(7403): p. 353-60. 
67. 
Peters, B. and A. Sette, Generating quantitative models describing the sequence specificity of 
biological processes with the stabilized matrix method. BMC Bioinformatics, 2005. 6: p. 132. 
68. 
Lundegaard, C., et al., NetMHC-3.0: accurate web accessible predictions of human, mouse and 
monkey MHC class I affinities for peptides of length 8-11. Nucleic Acids Res, 2008. 36(Web 
Server issue): p. W509-12. 
69. 
Hoof, I., et al., NetMHCpan, a method for MHC class I binding prediction beyond humans. 
Immunogenetics, 2009. 61(1): p. 1-13. 
70. 
Nielsen, M., et al., The role of the proteasome in generating cytotoxic T-cell epitopes: insights 
obtained from improved predictions of proteasomal cleavage. Immunogenetics, 2005. 57(1-2): p. 
33-41. 
71. 
Dannull, J., et al., Prostate stem cell antigen is a promising candidate for immunotherapy of 
advanced prostate cancer. Cancer Res, 2000. 60(19): p. 5522-8. 
72. 
Prazeres, D.M., et al., Purification of plasmids for gene therapy and DNA vaccination. Biotechnol 
Annu Rev, 2001. 7: p. 1-30. 
73. 
Manoj, S., L.A. Babiuk, and S. van Drunen Littel-van den Hurk, Approaches to enhance the 
efficacy of DNA vaccines. Crit Rev Clin Lab Sci, 2004. 41(1): p. 1-39. 
74. 
Donnelly, J., K. Berry, and J.B. Ulmer, Technical and regulatory hurdles for DNA vaccines. Int J 
Parasitol, 2003. 33(5-6): p. 457-67. 
75. 
Scheerlinck, J.P., et al., In vivo electroporation improves immune responses to DNA vaccination 
in sheep. Vaccine, 2004. 22(13-14): p. 1820-5. 

Washington University School of Medicine 
Protocol Date 08/03/23, A5 
Siteman Cancer Center 
Pancreatic Cancer SPORE 
Neoantigen DNA vaccines  
 
Page 60 of 69 
76. 
Babiuk, S., et al., Electroporation improves the efficacy of DNA vaccines in large animals. 
Vaccine, 2002. 20(27-28): p. 3399-3408. 
77. 
Luxembourg, A., et al., Enhancement of immune responses to an HBV DNA vaccine by 
electroporation. Vaccine, 2006. 24(21): p. 4490-3. 
78. 
Jaramillo, A., et al., Identification of HLA-A3-restricted CD8+ T cell epitopes derived from 
mammaglobin-A, a tumor-associated antigen of human breast cancer. Int.J.Cancer, 2002. 102(5): 
p. 499. 
79. 
Jaramillo, A., et al., Recognition of HLA-A2-restricted mammaglobin-A-derived epitopes by CD8+ 
cytotoxic T lymphocytes from breast cancer patients. Breast Cancer Res.Treat., 2004. 88(1): p. 
29. 
80. 
Manna, P.P., et al., Generation of CD8+ cytotoxic T lymphocytes against breast cancer cells by 
stimulation with mammaglobin-A-pulsed dendritic cells. Breast Cancer Res.Treat., 2003. 79(1): p. 
133. 
81. 
Narayanan, K., et al., Response of established human breast tumors to vaccination with 
mammaglobin-A cDNA. J.Natl.Cancer Inst., 2004. 96(18): p. 1388. 
82. 
Hobeika, A.C., et al., Enumerating antigen-specific T-cell responses in peripheral blood: a 
comparison of peptide MHC Tetramer, ELISpot, and intracellular cytokine analysis. J 
Immunother, 2005. 28(1): p. 63-72. 
83. 
Andersen, R.S., et al., Parallel detection of antigen-specific T cell responses by combinatorial 
encoding of MHC multimers. Nat Protoc, 2012. 7(5): p. 891-902. 
84. 
Cheung, R.K. and P.J. Utz, Screening: CyTOF-the next generation of cell detection. Nat Rev 
Rheumatol, 2011. 7(9): p. 502-3. 
85. 
Westfall, P.H. and S.S. Young, Resampling-Based Multiple Testing: Examples and Methods for 
p-Value Adjustment. 1993, New York: John Wiley & Sons, Inc. 

Washington University School of Medicine 
Protocol Date 08/03/23, A5 
Siteman Cancer Center 
Pancreatic Cancer SPORE 
Neoantigen DNA vaccines  
 
Page 61 of 69 
20 
APPENDICES 
 Abbreviations 
AE 
Adverse Event 
CBER 
Center for Biologics Evaluation and Research 
CFR 
Code of Federal Regulations 
CPA 
Cooperative Project Assurance 
CR 
Complete response 
CRA 
Clinical Research Associate 
CRF 
Case Report Form 
CT 
Computed Tomography 
CTCAE 
Common Terminology Criteria for Adverse Events 
CTEP 
Cancer Therapy Evaluation Program 
DNA 
Deoxyribonucleic acid 
DCTD 
Division of Cancer Treatment and Diagnosis 
ELISPOT 
Enzyme-linked immunospot assay 
ECOG 
Eastern Cooperative Oncology Group 
FDA 
Food and Drug Administration 
FWA 
Federal-wide Assurance 
GLP 
Good Laboratory Practice 
GMP 
Good Manufacturing Practice 
GPC 
Gel permeation chromatography 
HIPAA 
Health Insurance Portability and Accountability Act 
HLA 
Human leukocyte antigen 
HRPO 
Human Research Protection Office (Institutional Review Board at WUSM) 
IBC 
Institutional Biosafety Committee 
ICH 
International Conference of Harmonization 
IDB 
Investigational Drug Branch 
IRB 
Institutional Review Board 
LD 
Longest diameter 
MPA 
Multiple Project Assurance 
MRI 
Magnetic Resonance Imaging 
NCI 
National Cancer Institute 
NIH 
National Institutes of Health 
OBA 
Office of Biotechnology Activities 
OHRP 
Office for Human Research Protection 
PBMC 
Peripheral blood mononuclear cell 
PD 
Progressive disease 
PHI 
Protected Health Information 
PR 
Partial response 
RAC 
Recombinant DNA Advisory Committee 
RECIST 
Response Evaluation Criteria in Solid Tumors 
QASMC 
Quality Assurance and Safety Monitoring Committee 
SAE 
Serious Adverse Event 
SAS 
Statistical Analysis System; Analytical software from the SAS Institute, Cary, NC 
SCC 
Siteman Cancer Center 
SCIP 
Siteman Cancer Information Portal 
SD 
Stable disease 
SD 
Standard Deviation 
THF 
Tetrahydrofuran 
UPN 
Universal Product Number 
WUSM 
Washington University School of Medicine 
WUSTL 
Washington University in Saint Louis 

Washington University School of Medicine 
Protocol Date 08/03/23, A5 
Siteman Cancer Center 
Pancreatic Cancer SPORE 
Neoantigen DNA vaccines  
 
Page 62 of 69 
 ECOG/Zubrod performance status scale 
ECOG/Zubrod Score
Performance Status
0
Asymptomatic
1
Symptomatic, fully ambulatory
2
Symptomatic, in bed < 50% of the day
3
Symptomatic, in bed > 50% of the day but not bedridden
4
Bedridden
5
Dead
 
 National Cancer Institute Common Terminology Criteria for Adverse Events 
This study will collect adverse events using the NCI Common Terminology Criteria for Adverse Events 
v4.0 (CTCAE), if applicable. The CTCAE provides a descriptive terminology that is to be used for adverse 
event reporting. A grading (severity) scale is also provided in the CTCAE for each adverse event term. An 
electronic version of the CTCAE may be accessed through the web at http://ctep.cancer.gov. 
Alternatively, a full copy is available from the principal investigator.  
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

Washington University School of Medicine 
Protocol Date 08/03/23, A5 
Siteman Cancer Center 
Pancreatic Cancer SPORE 
Neoantigen DNA vaccines  
 
Page 63 of 69 
20.4 APPENDIX A: 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 
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 

Washington University School of Medicine 
Protocol Date 08/03/23, A5 
Siteman Cancer Center 
Pancreatic Cancer SPORE 
Neoantigen DNA vaccines  
 
Page 64 of 69 
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. 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

Washington University School of Medicine 
Protocol Date 08/03/23, A5 
Siteman Cancer Center 
Pancreatic Cancer SPORE 
Neoantigen DNA vaccines  
 
Page 65 of 69 
20.5 APPENDIX B: Reporting Timelines 
 
Expedited Reporting Timelines 
Event 
HRPO 
QASMC 
FDA 
IBC 
Ichor Medical Systems 
Serious AND 
unexpected suspected 
adverse reaction 
 
 
Report no later than 15 
calendar days after it is 
determined that the 
information qualifies for 
reporting 
Report at time of 
HRPO submission; 
IBC will retrieve 
information from 
myIRB and will 
follow up with study 
team if needed 
Since the TDS-IM device 
may be involved with the 
occurrence of adverse 
events, Ichor Medical 
Systems will be included 
in the Adverse Event 
reporting plan. Reports to 
Ichor for SAEs or for 
adverse events with 
possible relationship to the 
device should follow the 
timing for reporting to the 
WUSM Human Research 
Protection Office and 
should be directed to 
Drew Hannaman at 
dhannaman@ichorms.com 
Unexpected fatal or 
life-threatening 
suspected adverse 
reaction 
 
 
Report no later than 7 
calendar days after initial 
receipt of the information 
 
 
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 
 
 
 
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. 
 
 
 
 
A series of minor 
Report within 10 working days.    
 
 
 

Washington University School of Medicine 
Protocol Date 08/03/23, A5 
Siteman Cancer Center 
Pancreatic Cancer SPORE 
Neoantigen DNA vaccines  
 
Page 66 of 69 
Expedited Reporting Timelines 
Event 
HRPO 
QASMC 
FDA 
IBC 
Ichor Medical Systems 
deviations that are 
being reported as a 
continuing 
noncompliance 
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 
 
 
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. 
 
 
 
 
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 
 
 
 
 

Washington University School of Medicine 
Protocol Date 08/03/23, A5 
Siteman Cancer Center 
Pancreatic Cancer SPORE 
Neoantigen DNA vaccines  
 
Page 67 of 69 
Expedited Reporting Timelines 
Event 
HRPO 
QASMC 
FDA 
IBC 
Ichor Medical Systems 
withdrawn, report at continuing 
review. 
 
Routine Reporting Timelines 
Event 
HRPO 
QASMC 
FDA 
Ichor Medical Systems 
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. 
 
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. 
 
 
 
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. 
 
 
 
 
 
 
 
 
 

Washington University School of Medicine 
Protocol Date 08/03/23, A5 
Siteman Cancer Center 
Pancreatic Cancer SPORE 
Neoantigen DNA vaccines  
 
Page 68 of 69 
Expedited Reporting Timelines for Secondary Sites 
Event 
WU (Coordinating Center) 
Local IRB 
FDA 
Ichor Medical Systems 
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 Ichor 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. 
 
 

Washington University School of Medicine 
Protocol Date 08/03/23, A5 
Siteman Cancer Center 
Pancreatic Cancer SPORE 
Neoantigen DNA vaccines  
 
Page 69 of 69 
20.6 APPENDIX C: Washington University Unanticipated Problem Reporting 
Cover Sheet 
 
SAE COVER SHEET- Secondary Site Assessment 
 
Washington University HRPO#:  
Sponsor-Investigator:  
Subject Initials:   
Subject ID:  
Treating MD:  
Treating Site:  
EVENT TERM:  
Admission 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