Phase I Trial of Intravenous Administration of Vesicular Stomatitis Virus Genetically Engineered to Express Thyroidal Sodium Iodide Symporter (NIS) and Human Interferon Beta (hIFNb), in Patients With Metastatic or Recurrent Endometrial Cancer
Phase: PHASE1
Conditions: Metastatic Endometrial Carcinoma, Recurrent Endometrial Adenocarcinoma, Recurrent Endometrial Carcinoma, Recurrent Endometrial Clear Cell Adenocarcinoma, Recurrent Endometrial Endometrioid Adenocarcinoma, Recurrent Endometrial Mixed Cell Adenocarcinoma, Recurrent Endometrial Serous Adenocarcinoma, Recurrent Endometrial Undifferentiated Carcinoma, Recurrent Uterine Corpus Carcinosarcoma, Stage IV Uterine Corpus Cancer AJCC v7
Sponsor: Mayo Clinic

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Mayo Clinic Cancer Center 
MC1562 Phase I Trial of Intravenous Administration of Vesicular Stomatitis Virus Genetically 
Engineered to Express Thyroidal Sodium Iodide Symporter (NIS) and Human Interferon beta 
(hIFNβ), in Patients with Metastatic or Recurrent Endometrial Cancer 
 
Study Chair: 
 
 
 
 
Study Co-Chairs: 
 
 
 
Study Co-Investigators: 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Statisticians: 
 
 
√Study contributor(s) not responsible for patient care 
 
Drug Availability 
Supplied Investigational Agents: VSV-IFN-NIS (IND# 16811) 
 
Document History 
Effective Date 
 
 
 
Activation 
15Sep2017 
 
MCCC Amendment 7 
pending 
MCCC Amendment 1 
01May2018 
 
 
 
MCCC Amendment 2 
22Aug2018 
 
 
 
MCCC Amendment 3 
08Feb2019 
 
 
 
MCCC Amendment 4 
14May2019 
 
 
 
MCCC Amendment 5 
24Jul2019 
 
 
 
MCCC Amendment 6 
24Nov2020 
 
 
 
 
 

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Protocol Resources 
Questions: 
Contact Name: 
Patient eligibility*, test schedule, 
treatment delays/interruptions/adjustments, 
dose modifications, adverse events, 
forms completion and submission 
 
 
 
 
Clinical Study Coordinator 
 
 
 
Clinical Data Coordinator 
 
 
  
 
Protocol document, consent form, 
regulatory issues 
 
 
 
 
Laboratory Contacts for Correlative Studies 
 
 
 
 
 
 
Serious Adverse Event Reporting  
 
 
 
  
*No waivers of eligibility allowed 
 
 

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Table of Contents 
Phase I Trial of Intravenous Administration of Vesicular Stomatitis Virus Genetically Engineered to 
Express Thyroidal Sodium Iodide Symporter (NIS) and Human Interferon beta (hIFNβ), in Patients with 
Metastatic or Recurrent Endometrial Cancer ................................................................................................ 1 
Protocol Resources ........................................................................................................................................ 2 
Table of Contents .......................................................................................................................................... 3 
Schema for Group A VSV-hIFNβ-NIS alone – PERMANENTLY CLOSED ............................................. 4 
Schema for Group B VSV-hIFNβ-NIS with Ruxolitinib .............................................................................. 5 
1.0 
Background ................................................................................................................................... 6 
2.0 
Goals ........................................................................................................................................... 15 
3.0 
Patient Eligibility ........................................................................................................................ 16 
4.0 
Test Schedule(s) .......................................................................................................................... 18 
5.0 
Grouping Factors ........................................................................................................................ 20 
6.0 
Registration/Randomization Procedures ..................................................................................... 20 
7.0 
Protocol Treatment ...................................................................................................................... 22 
8.0 
Dosage Modification Based on Adverse Events ......................................................................... 29 
9.0 
Ancillary Therapy ....................................................................................................................... 30 
10.0 
Adverse Event (AE) Reporting and Monitoring .......................................................................... 35 
11.0 
Treatment Evaluation. ................................................................................................................. 44 
12.0 
Descriptive Factors ...................................................................................................................... 48 
13.0 
Treatment/Follow-up Decision at Evaluation of Patient .............................................................. 49 
14.0 
Pharmacologic/Ancillary studies ................................................................................................ 50 
15.0 
Drug Information ........................................................................................................................ 54 
16.0 
Statistical Considerations and Methodology ............................................................................... 59 
17.0 
Pathology Considerations ............................................................................................................ 64 
18.0 
Records and Data Collection Procedures .................................................................................... 67 
19.0 
Budget Considerations ................................................................................................................ 68 
20.0 
References ................................................................................................................................... 69 
Appendix I 
ECOG Performance Status .................................................................................................. 73 
Appendix II 
New York Heart Association Classification of Congestive Heart Failure .......................... 74 
Appendix III 
Child-Pugh Score ............................................................................................................ 75 
Appendix IV 
Study Drug Diary ............................................................................................................ 76 

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Schema for Group A VSV-hIFNβ-NIS alone – PERMANENTLY CLOSED 
NOTE: Group A is permanently closed as of 10Jan2023. 
Prior to discussing protocol entry with the patient, call the Mayo Clinic Research Site Management 
Office 
) to confirm study status and ensure that a place on the protocol is currently 
available to the patient 
 
 
Registration 
(See Section 6.0) 
 
 
 
 
↓ 
 
 
 
Archived tumor or research biopsy 
and optional baseline NIS imaging 
 
 
 
↓ 
 
 
 
 
1 dose of VSV-hIFNβ-NIS 
(at assigned dose) 
 
 
 
 
↓ 
 
 
 
Optional NIS imaging Day 2 after VSV-
hIFNβ-NIS 
(and up to 2 other scans if previous imaging 
data are positive) 
Biopsy of NIS-lit areas may occur after any 
NIS imaging 
 
 
↓ 
 
 
Imaged-guided tumor biopsy on Day 29 
 
 
 
↓ 
 
 
 
 
Observation 
 
Disease progression, 
refusal, or alternate 
therapy at any time 
 
 
↓ 
 
↓ 
 
 
Event Monitoring 
 
Event Monitoring 
 
Cycle = 29 days (one cycle) 
NOTE: Groups enroll independently  
 
Generic name: VSV-hIFNβ-NIS (recombinant vesicular stomatitis 
virus carrying the human NIS and IFNβ genes) 
Brand name(s): NA 
Mayo Abbreviation: VSV-HIFNB-NIS 
Availability: Mayo Clinic Viral Vector Production Lab 
 

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Schema for Group B VSV-hIFNβ-NIS with Ruxolitinib 
Prior to discussing protocol entry with the patient, contact the Mayo Clinic Research Site 
Management Office (
) to confirm study status and ensure that a place on the 
protocol is currently available to the patient 
 
 
Registration 
(See Section 6.0) 
 
 
 
 
↓ 
 
 
 
Archived tumor or research biopsy 
and optional baseline NIS imaging 
 
 
 
↓ 
 
 
 
Start ruxolitinib 3 days before VSV 
administration and continue for up to 9 days 
(12 days total) 
1 dose of VSV-hIFNβ-NIS 
(at assigned dose) 
 
 
 
↓ 
 
 
 
Optional NIS imaging Day 2 after VSV-
hIFNβ-NIS 
(and up to 2 other scans if previous imaging 
data are positive) 
Biopsy of NIS-lit areas may occur after any 
NIS imaging 
 
 
↓ 
 
 
Imaged-guided tumor biopsy on Day 29 
 
 
 
↓ 
 
 
 
 
Observation 
 
Disease progression, 
refusal, or alternate 
therapy at any time 
 
 
↓ 
 
↓ 
 
 
Event Monitoring 
 
Event Monitoring 
Cycle = 29 days (one cycle) 
NOTE: Groups enroll independently 
Generic Name 
Brand 
Name(s) 
Mayo 
Abbreviation 
Availability 
VSV-hIFNβ-NIS (recombinant 
vesicular stomatitis virus carrying the 
human NIS and IFNβ genes) 
NA 
VSV-HIFNB 
Mayo Clinic Viral Vector 
Production Laboratory 
Ruxolitinib 
Jakafi® 
 
 

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1.0 
Background 
1.1 
Introduction 
Vesicular stomatitis virus (VSV) is a negative strand RNA virus that belongs to the family 
Rhabdoviridae (order Mononegavirales) [1-3]. It can be divided into two antigenically 
distinct species: VSV-Indiana and VSV-New Jersey. Both species can infect insects and 
mammals and the New Jersey strain is an important pathogen for cattle and swine in the 
United States. Infections in these mammals causes oral and tongue vesicles that can be 
mistaken for the much more serious hand-foot and mouth disease. Human exposure is rare 
and is predominantly found in laboratory workers who handle the virus, veterinarians and 
farm workers who are exposed to infected livestock. Infection in humans is either clinically 
silent or presents as a mild influenza-like illness with fever, headache, general malaise, 
myalgia, nausea and vomiting. About a quarter of patients may also have transient vesicles 
in their mouths or on the lips and nose. Treatment is symptomatic and the illness resolves 
spontaneously as the immune system clears the virus [3, 4]. There is one report linking VSV 
with encephalitis, in a young child [5]. Mammals infected with VSV do not exhibit a 
viremic phase [6]and infection is via arthropod transmission of the virus. The main species 
responsible for transmission is the black fly (Simulium vittatum) although the sandfly 
(Lutzomyia sp) is also implicated. Mammal to mammal transmission occurs rarely in swine 
as a consequence of vesicle contact [7], however, human to human transmission, to our 
knowledge does not occur. Hence the risk of spread of the virus from patients to their care 
providers or family is highly unlikely. VSV has broad oncolytic activity including activity in 
breast, colon, multiple myeloma, as well as ovarian and endometrial cancer. The virus preferen-
tially replicates in tumor cells due to defects in interferon signaling present in such cells. 
Viral infection leads to rapid amplification of the virus with death of infected cells by lysis. 
1.2 
VSV-hIFNβ-NIS description 
VSV-hIFNβ-NIS is a recombinant, replication competent virus based on the Indiana 
strain of VSV [8, 9]. The VSV genome codes for 5 proteins: the nucleoprotein (N), 
phospho-protein (P), matrix (M), membrane glycoprotein (G) and large (L) those are 
required for the structure and life cycle of the virus. VSV is a bullet shaped virus with a 
phospholipid bilayer envelope derived from the infected cell. The envelope contains the G 
glycoprotein res-ponsible for binding and entry into target cells while the matrix protein 
(M) is a structural protein lying below the envelope and is essential for viral assembly. 
The G protein confers broad cell tropism since the receptor for this glycoprotein is a 
phospholipid that is ubi-quitously expressed by cells. The N and P proteins form 
complexes with the RNA genome while the L protein is the viral RNA dependent RNA 
polymerase. Infected cells rapidly amplify the virus that ultimately kills cells by lysis. 
VSV-hIFNβ-NIS has two additional human genes cloned into its genome: the gene for 
human interferon β (hIFNβ) downstream of the M gene and the gene coding for the 
human sodium iodide symporter (NIS) down-stream of the gene for the G protein. The 
recombinant virus remains lytic to susceptible cells and can be grown to very high titers 
under Good Manufacturing Practice (GMP) conditions. VSV is very sensitive to the effect 
of interferon. Indeed, one of the functions of the M protein is to block the extranuclear 
transport of mRNA coding for interferons that will block viral replication. Non-
transformed cells produce and respond to IFN normally, but many tumor cells have lost 
the capacity to either produce or respond to interferon. Thus, interferon production by 
tumor cells infected with VSV-hIFNβ-NIS will protect normal cells but not tumor cells. 
NIS expression enables tumor cells infected with the virus to concentrate various isotopes 
such as 99mTc pertechnetate that allows non-invasive, and repeated imaging of the 
biodistribution of virus infected cells (as with MV-NIS) [10] [11, 12] 

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1.3 
VSV-hIFNβ-NIS: Mechanism of tumor specificity 
VSV is an RNA virus that completes its life cycle in the cytoplasm of infected cells. The 
receptor utilized for binding and entry to cells is recently identified to be from the low 
density lipid LDL receptor (LDLR) family that is ubiquitous in mammalian cells. This 
explains the use of the G glycoprotein to pseudotype many viruses to widen their cellular 
tropism. Normal cells respond to RNA virus infection by the production of interferons. 
These proteins are secreted and work in an autocrine and paracrine fashion to halt protein 
synthesis in cells so that viral replication will be blocked [13] . IFN binds to specific cell 
surface receptors that have tyrosine kinase activity. Once phosphorylated, the 
cytoplasmic parts of the receptors serve as docking sites for JAK1-2 and TYK2 that in 
turn recruit STAT1-2 and in combination with IRF-9 to form the heterotrimeric complex 
known as IFN-stimulated gene factor 3 (ISGF3) which migrates to the nucleus where it 
promotes transcription of specific genes that have IFN-stimulated response elements in 
their promoter region. In addition, phosphorylation of eIF2α blocks protein synthesis in 
cells, effectively inhibiting viral replication [2, 8]. In contrast to normal cells, many 
tumor cells have defects in IFN production, or response to the cytokine making them 
particularly susceptible to VSV replication and lysis. For this reason, VSV has been 
explored as a novel oncolytic agent by various groups [1, 8, 9]. IFN produced by infected 
cell may also inhibit angiogenesis in the tumor microenvironment, and provide another 
mechanism of tumor control. VSV has been shown to induce oncolysis in breast, cervical, 
pancreatic, hepatic, colon, prostate cancer, melanoma, and multiple myeloma as well as 
endometrial and ovarian cancer. [4, 14-19]. 
The broad tumor specificity of VSV is due to the fact that the virus does not depend on a 
specific molecular defect within tumor cells to kill them. In fact, VSV can kill tumor cells 
regardless of p53, Ras or c-Myc expression or status [20].  
One of the most important pathways through which VSV induces oncolysis is the IFN 
pathway, which stimulates transcription of p53 which subsequently promotes apoptosis in 
the cells. VSV-mp53 and VSV-M(mut)-mp53 express high levels of functional p53 and 
retain the ability to lyse transformed versus normal cells [21]. p53 has potent antitumor 
properties; in a mouse model of metastatic mammary adenocarcinoma, a single 
inoculation of VSV-M-mp53 resulted in increased survival compared to those that did not 
receive virus [22].  
1.4 
VSV-hIFNβ-NIS Radioactive iodide uptake 
Cells infected with VSV-hIFNβ-NIS express the thyroidal sodium iodide symporter 
(NIS). NIS is a 643 amino acid glycoprotein that is physiologically expressed on the 
plasma membrane of thyroid epithelial cells and to a lesser extent in the salivary glands, 
stomach and kidney. The protein concentrates isotopes such as 99mTc pertechnetate or 123I 
and is the basis of successful therapy of advanced thyroid cancer with the beta particle 
emitter 131I [23-25]. Tumor cells that express NIS can concentrate various isotopes taken 
up by this protein, enabling both molecular imaging as well as therapy in some 
circumstances [10, 11, 26-29]. We have performed studies in C57BL/KaLwRij mice 
bearing syngeneic 5TGM1 subcutaneous tumors where animals are injected with a single 
dose of VSV-hIFNβ-NIS (108 TCID50) and imaging started the next day after injection 
of 99mTc pertechnetate (Figure 1). A distinct and measurable signal was detectable in all 
tumors from mice injected with VSV-murine IFNβ-NIS but not from controls. The 
strength of the signal increased in time as the virus spread within the tumor, reaching a 
peak at 2 days in most animals and then the signal decreased over the course of the next 4 
days, as the tumors were controlled by the spreading infection. Concomitant CT imaging 

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of the animals confirmed the significant reduction in tumor burden after this single
injection of the virus. It is important to note that these experiments are in
immunocompetent animals. Using volume of interest analysis, we calculated that at the
peak of NIS expression, the tumors concentrated ~15% of the injected isotope. Thus, NIS 
expression enables almost daily tracking of the biodistribution and degree of infection
within localized tumors.
1.5
Endometrial Cancer (EC)
EC is the most common gynecologic malignancy in the United States with an estimated 
47,130 new cases and 8,010 deaths from disease in 2012 [30]. While low-risk surgical 
stage I EC has a 5-year overall survival of 96% with surgical resection alone, 5-year 
overall survival for stage III and IV disease is 67% and 16%, respectively [31]. In 
addition, optimal therapy for high-risk early-stage and advanced-stage disease remains 
unclear. External beam radiotherapy [32-34], systemic chemotherapy [35, 36], combined 
chemotherapy and radiation [37], and most recently, biologics [34, 35] have been and 
continue to be investigated as adjuvant therapies for EC after surgical staging. Despite 
nearly 3 decades of randomized controlled trials of adjuvant therapy, 5-year overall 
survival in metastatic EC continues to decline [31, 38, 39]. Patients with recurrent disease 
represent a heterogeneous sub-group of patients and more than half of recurrences will 
develop within 2 and 3 years after primary treatment [40]. 
While isolated EC vaginal recurrences can be salvaged with pelvic radiation and 
brachytherapy [40][41], multisite and distant recurrence often portends death from 
disease [31]. Numerous systemic cytotoxic therapies have been investigated for recurrent 
EC [42-46], with response rates ranging from 0% for oral etoposide [46] to 27.3% for 
single-agent paclitaxel [43]. However, the response rate for most single-agent 
chemotherapies investigated in patients with recurrent disease remains in the single-digit 
percentages [40, 45]. Additionally, in the past decade, biologic agents have emerged, and 
their activity as single agents in the patients with recurrent EC has been similar to that of 
cytotoxic agents, with response rates ranging from 3.3% for lapatinib [47] to 13.5% for 
bevacizumab [39]. And while progestin therapy for recurrent EC may extend 
progression-free survival, it does not improve overall survival [48].
Taken together, the substantial risk of recurrence in advanced-stage EC and the modest 
response to systemic therapies in patients with recurrence suggest that novel approaches 
are imperative in mitigating the high risk of death from advanced-stage and recurrent EC.
Figure 1: Left panel of the same mouse with flank 5TGM1 tumors given one IV 
dose of VSV-IFN-NIS and imaged daily.  Right panels show quantitative analysis 
of isotope uptake in the flank tumor (n=5 mice). 

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1.6 
Oncolytic virotherapy clinical trials in EC 
It has been a century since the first published account of viral oncolytic activity in 
gynecologic cancer when an advanced cervical cancer regressed in response to rabies 
vaccination [49].  At present, EOC is the gynecologic malignancy that has garnered the 
most focus regarding oncolytic virus therapy. Measles virus, vaccinia, herpes simplex, 
reovirus, several adenoviruses, and VSV all have shown activity against ovarian cancer in 
preclinical cancer models [50, 51], and EOC clinical trials have been performed using 
measles, herpes simplex, reovirus, and adenoviruses [52-54]. 
Oncolytic virotherapy in EC, however, is only just emerging. A small cohort of 3 cases of 
recurrent EC treated on a phase I trial with intraperitoneal Ad5.SSTR/TK.RGD, an 
infectivity-enhanced adenovirus was promising, with 2 recurrent ECs maintaining stable 
disease [32]. To date, there exist no other published cases of EC treated with oncolytic 
virus therapy. 
1.7 
Rationale for Testing VSV-hIFNβ-NIS in EC  
Drs. KahWhye Peng, Stephen Russell, and colleagues have developed a genetically 
engineered VSV that expresses the human IFN-beta gene. We have shown through a 
series of experiments that this VSV-hIFNβ virus replicates efficiently in both type I (Fig 
2) and type II human EC cell lines (Fig 3). Moreover, we have shown that the VSV-
hIFNß virus is cytotoxic to both type I (Fig 4) and type II human EC cell lines (Fig 5) and 
improves overall survival among mice with AN3CA cell line xenografts (Fig 6). 
Additionally, high copy numbers of VSV are observed in xenografted tumors after IV 
administration of VSV-hIFNβ (Figure 7) and low in normal tissues, supporting the 
selective infectivity of EC by VSV. 
 
 
 
 
 
 
 
Figure 2 
Figure 3 
Figure 4 
Figure 5 

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VSV-hIFNβ is being tested in a phase I clinical trial for patients with hepatocellular 
carcinoma at Mayo Clinic in Arizona (PI Mitesh Borad; Clinical trials.gov identifier 
NCT01628640).  
Given the high mortality associated with advanced-stage and recurrent EC, the efficacy 
limitations of currently available therapies, and the promising antitumor potency of VSV, 
we propose a first-in-human study of IV VSV-hIFNβ-NIS in women with metastatic, 
incurable EC.  
1.8  
p53 in EC  
While VSV appears to cause apoptosis mainly through the IFN pathway, the p53 
pathway, a critical pathway in EC carcinogenesis, also plays a critical role in VSV 
oncolysis [55]. Differing histologic appearance and clinical behavior has led to the 
separation of EC into two sub-groups: type I includes low-grade, endometrioid histology 
which typically has a good prognosis with >75% of cases presenting at an early stage. 
Type II EC includes serous and clear cell histologies and a higher proportion present with 
metastatic disease and worse prognosis than type I EC [56]. p53 plays a role in EC 
carcinogenesis, specifically within the aggressive type II EC which overexpresses p53 
[57]. In fact, p53 expression is a prognostic biomarker with potential clinical utility as it 
is associated with reduced survival in patients with EC. p53 not only loses its genome 
stabilizing and tumor suppression activities, but also exhibits oncogenic function in 
cancer cells [30, 58-60].  
1.9a 
Safety of VSV in humans 
VSV is an important pathogen in the field of veterinary medicine, since its symptoms can 
be mistaken for those of foot and mouth disease. The virus infects swine, cattle and horses 
and most humans in the United States have not been exposed to the virus although human 
infection is common in Central America. In the United States, adults that are infected 
typically work with farm animals or are in the veterinary profession. Therefore, the vast 
majority of humans do not have immunity to the virus [4]. Mammals infected with VSV 
exhibit a short lived low level viremic phase [6] and infection is via arthropod 
transmission of the virus. The main species responsible for transmission is the black fly 
(Simulium vittatum) although the sandfly (Lutzomyia sp) is also implicated. Mammal to 
mammal transmission occurs rarely in swine as a consequence of direct contact with 
vesicles containing the virus [8][7]. However, human to human transmission, to our 
knowledge does not occur. Hence the risk of spread of the virus from patients to their care 
Figure 6 
Figure 7 

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providers or family is highly unlikely. Humans infected with VSV are usually 
asymptomatic or have a mild, flu-like illness that resolves spontaneously within a few 
days and only requires symptomatic therapy.  
The most serious complication of VSV infections have included encephalitis that has been 
reported in a single 3 year old patient from Panama [6][5]. Additionally, a male patient 
that received the recombinant VSV that expresses the human interferon β gene (VSV-
hIFNβ) in a phase I clinical trial at Mayo Clinic Arizona (PI Mitesh Borad; Clinical 
Trials.gov identifier NCT01628640), where the virus is being administered by image 
guided intratumoral injection into a liver tumor nodule, died after receiving VSV-hIFNβ. 
He had extensive cancer involvement of the liver and experienced severe liver injury 
following injection of VSV-hIFNβ (personal communication with study PI). 
1.9b 
Therapeutic efficacy and noninvasive imaging of VSV-hIFNβ-NIS 
VSV derivatives have potent and rapid oncolytic activity against multiple myeloma. VSV 
(D51)-NIS is a virus with a mutation in the M protein that increases its susceptibility to 
IFN [10]. This virus was engineered to express NIS and tested against the 5TGM1 
myeloma model. A single injection of VSV(D51)-NIS (5 × 108 TCID50) was able to 
significantly slow down the growth of syngeneic subcutaneous 5TGM1 tumors in 
immunocompetent animals [49]. This slow down was translated into a significant 
prolongation of survival. In a second study, the same virus also produced a response in an 
orthotopic model of myeloma: a single injection of the virus reduced the paraprotein level 
in treated mice compared to controls and also prolonged survival [49]. Despite VSV-
hIFNβ-NIS being a safer virus due to expression of IFNβ by infected cells, the virus 
retains its potent oncolytic potency against multiple myeloma cell lines and primary cells. 
Indeed, a single dose of VSV-hIFNβ-NIS was able to control 5TGM1 tumors growing 
subcutaneously in syngeneic mice or as an orthotopic model. The biodistribution of the 
virus infected population can be serially monitored by 99mTc pertechnetate administration 
and planar and SPECT/CT imaging [49] or 18F-TFB administration with PET/CT or 
PET/MR imaging. Moreover, due to NIS expression, therapy with the beta emitting 
isotope 131I led to an even greater improvement in disease control and survival due to the 
bystander effect of the electrons emitted by the decay of this isotope [29, 49].  
1.9c 
Preclinical data: VSV-hIFNβ-NIS toxicity and biodistribution studies in mice and 
canine 
The activity of interferon beta is species specific, hence human IFNβ is not biologically 
active in mice or canines. For our toxicology and efficacy studies, we have also generated 
viruses that express the murine (VSV-mIFNβ-NIS) or canine interferon beta genes (VSV-
cIFNβ-NIS) in place of hIFNβ (Figure 8). 
 
 
Figure 8: Schematic representation of the full length infectious clones of VSV-IFNβ-NIS 
encoding species specific IFNb cDNA. 
IFNβ
NIS
VSV-IFNβ-NIS
Human IFNβ
Murine IFNβ
Canine IFNβ

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Dose response efficacy study of VSV-mIFN-NIS in immunocompetent and irradiated 
SCID mice. C57BL/KaLwRij mice with subcutaneous 5TGM1 syngeneic myeloma 
tumors received one intravenous dose of saline or 105, 106, 107 or 108 TCID50 VSV-
mIFNβ-NIS via the tail vein. The virus significantly extended survival of mice in all 
treatment groups, and there was a clear dose response in relation to survival of animals 
and duration of tumor response, where the highest dose group showed the best response 
data. CB17 SCID mice were irradiated at 150 rads before implantation of human KAS6/1 
myeloma tumor cells. When the subcutaneous flank tumors were established, mice 
received one intravenous dose of VSV-mIFNβ-NIS (105, 106, 107 or 108 TCID50). Mice in 
the virus treated groups responded to the virus therapy with regression of tumors and 
extended survival compared to the saline treatment group. Importantly, no toxicity, in 
particular no clinical signs of neurotoxicity, was observed in any of the treated animals at 
the top doses used in these (~20 g) mice. 
Human Equivalent Dose = [70/0.02] * 108 TCID50 = 3.5 x 1011 TCID50/70 kg human 
• 
assume 20g mouse 
Dose response efficacy and safety study of VSV-hIFNβ-NIS and VSV-cIFNβ-NIS in 
research hounds and companion canines with spontaneous cancer. A rapid dose-
escalation study in purpose-bred Beagle dogs delineated adverse event (AE) profile and 
Maximum Tolerated Dose (MTD) following systemic VSV-hIFNβ-NIS administration, 
indicating 1010 TCID50 (per 0.5m2) was well tolerated in normal dogs. At 1011 TCID50, 
severe adverse events occurred and corollary evaluation indicated the primary dose 
limiting toxicity was hepatotoxicity. A gradual dose escalation study in pet dogs with 
diagnosed hematologic malignancies is ongoing at the University of Tennessee at a 
starting dose of 1010 TCID50 with two-fold dose escalations treating 3 dogs at each dose 
level, alternating between VSV-hIFNβ-NIS and VSV-cIFNβ-NIS. A total of 5 dogs were 
treated with VSV-hIFNβ-NIS and 4 with VSV-cIFNβ-NIS. Analysis of virus shedding 
suggested low to no detection of virus genomes in buccal swabs or urine. Analysis of 
infectious virus recovery from urine and buccal swab samples were mostly negative, 
except for suspected contaminated samples. There was no significant treatment 
associated toxicity (mild and transient fever, and lymphopenia). All treated dogs had 
transient elevations in LFTs. While most LFT elevations were mild, two dogs, Beasley 
and Roxie, had significantly elevated ALTs in the first week following VSV therapy. 
Beasley had an approximately 2X ULN (Upper level of normal) increase in ALT, a 
Grade 2 toxicity according to VCOG CTCAE. This elevation was transient and 
normalized by Day 7. Roxie’s ALT elevation was approximately 10X ULN, a Grade 3 
toxicity, which normalized by Day 7 also. Both were T-cell lymphoma bearing dogs 
treated with VSV-hIFN-NIS and had the most significant responses to systemic VSV 
therapy (transient response with decrease in tumor size). 
Human Equivalent Dose = [70/10] * 1010 TCID50 = 7 x 1010 TCID50/70 kg human 
• 
assume 10kg dog 
1.9d 
VSV-hIFNβ-NIS Translation 
A recombinant VSV that expresses the human interferon β gene (VSV-hIFNβ) is in 
clinical testing at Mayo Clinic Arizona where the virus is given by image guided 
intratumoral injection into a liver tumor nodule (PI: Dr. Mitesh Borad, IND#14347, 
Clinical Trials.gov identifier: NCT01628640). 
A patient enrolled in the Arizona study died after receiving VSV-hIFNβ. He had extensive 
cancer involvement of the liver and experienced severe liver injury following injection of 

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VSV-hIFNβ. It is not known what role the virus played in the liver effects because at the 
start of the trial the patient’s liver was filled with tumor and very little normal liver 
remained. 
As a precaution all patients will be monitored for 24 hours post-infusion in the Mayo 
Clinic Hospital –Methodist Campus. Additionally, daily assessments and laboratory tests 
(CBC, electrolytes, liver enzymes, and coagulation panel) will be performed for the first 
5 days after VSV administration to assess for liver toxicity and tumor lysis syndrome 
(TLS). 
VSV-hIFNβ-NIS is a similar virus with the added benefits of (i) in vivo molecular 
imaging and (ii) the potential to combine virotherapy with selective and localized 
radiation due to specific radioisotope uptake and concentration (131I).  
The current study is the first to use such a virus systemically in patients with EC. As such 
the trial has a number of unique features including: (i) VSV-hIFNβ-NIS enables us to 
track its biodistribution, replication and gene expression using non-invasive imaging, so 
we can determine the pharmacokinetics and pharmacodynamics of the virus in vivo; (ii) 
Unlike many other oncolytic viruses currently being tested in clinical trials, the patient 
population is not expected to have pre-existing anti-VSV immunity [5] that can neutralize 
the virus in its systemic journey to tumor sites. For many viruses, this has been a 
significant hurdle that impedes access of the virus to the tumor. Therefore, for the first 
time, the potential impact of an oncolytic virus on human cancer can be evaluated with a 
reasonable expectation of establishing an infection within the target tissue. (iii) With 
VSV, there is the possibility to administer high titers of replicating oncolytic virus, 
thereby improving the potential for tumor cell infection and oncolysis. In the current 
study, the safety and efficacy of VSV-hIFNβ-NIS alone will be determined and if 
effective, the potential to combine it with therapeutic radioisotopes will be determined in a 
subsequent trial. 
1.9e 
VSV-hIFNβ-NIS and ruxolitinib 
Tumor burden was limited in the initial phase 1 trial to constrain the potentially high 
levels of interferon beta production by infected cells. In order to abrogate this concern, 
we propose using ruxolitinib in conjunction with VSV-hIFNβ-NIS in a new group of 
patients with no tumor burden restriction. 
Ruxolitinib (Jakafi™) is an oral Janus-associated kinase 1 (JAK1) and JAK2 inhibitor 
used clinically for the treatment of polycythemia vera and myelofibrosis. Side effects of 
ruxolitinib include thrombocytopenia, anemia and neutropenia, but these are dose-
dependent and can be modulated by reducing the dose or withholding ruxolitinib. VSV-
hIFNβ-NIS infected cells express IFNβ. Binding of IFNβ to its receptor and subsequent 
activation of downstream interferon inducible antiviral genes is mediated via Jak1/Stat 
signaling. It has been shown that addition of ruxolitinib with VSV infection sensitizes 
VSV resistant cancer cells to virus infection (Escobar et al, 2013). Importantly, we 
demonstrated in preclinical studies that ruxolitinib can ameliorate toxicity associated with 
high circulating levels of interferon beta in Balb/c mice. When given orally to Balb/c 
bearing a highly VSV susceptible MPC-11 tumor, ruxolitinib is able to protect Balb/c 
mice from lethal toxicity associated with acute tumor lysis syndrome and high circulating 
levels of IFNβ after these tumor bearing mice were treated with an intravenous dose of 
VSV-mIFNβ-NIS. The antitumor activity of VSV-IFN-NIS was not negated and tumor 
growth was significantly inhibited compared to saline controls.  

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1.9f 
Summary 
This trial is two parallel phase I trials. Both treatment groups will follow the same dosing 
escalation schedule for the VSV-hIFNβ-NIS, will follow the same dose limiting toxicity 
(DLT) determination and will be analyzed separately. 
1.9g 
Hypotheses 
The primary hypotheses to be tested in this Phase I trial are as follows: 
1. VSV-hIFNβ-NIS administered intravenously (IV) to patients with metastatic and/or 
recurrent EC will selectively propagate in deposits of endometrial cancer cells 
throughout the body leading to tumor cell killing and a reduction in tumor burden. 
2. Longitudinal NIS imaging using Tc-99m pertechnetate and planar and SPECT/CT or 
TFB-PET enables detection of pharmacokinetics of viral replication in the measureable 
EC. 
3. Ruxolitinib administered to patients undergoing treatment with VSV-hIFNβ-NIS will 
be safe, enhance antitumor activity of VSV and protect patients from toxic effects 
associated with high IFNβ levels. 
 
 
 

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2.0 
Goals 
2.1 
Primary 
To evaluate the optimal dose schedule, safety and tolerability as measured by the 
incidence of significant toxicity of VSV-hIFNβ-NIS in immunocompetent patients with 
metastatic and/or recurrent EC. 
 
2.2 
Secondary 
2.21 
To determine the toxicity profile of VSV-hIFNβ-NIS (alone and in combination 
with ruxolitinib).  
2.22  
To determine the time course of viral gene expression and virus elimination, and 
the biodistribution of virally infected cells at various times points after infection 
with VSV-hIFNβ-NIS (alone and in combination with ruxolitinib) using Tc-99m 
pertechnetate planar and SPECT/CT or TFB-PET imaging. 
2.23 
To assess virus replication, viremia; viral shedding in urine and respiratory 
secretions; and virus persistence after IV administration of VSV-hIFNβ-NIS 
(alone and in combination with ruxolitinib). 
2.24 
To monitor humoral responses to the injected virus. 
2.25 
To estimate the tumor response rate and overall survival. 
 
2.3 
Correlative 
2.31 
To determine the pharmacokinetic (PK) profile of VSV-IFNβ-NIS in patients 
with EC by measurement of VSV-IFNβ-NIS in blood by reverse transcriptase 
polymerase chain reaction (RT-PCR). 
2.32 
To characterize the pharmacodynamics (PD) of VSV-IFNβ-NIS by way of 
measuring serum interferon-β and also VSV-RT-PCR of VSV-IFNβ-NIS listed 
above. 
2.33 
Assess CD8+ T cell (both general and VSV-IFNβ-NIS specific) and NK cell 
responses. 
2.34 
Gene expression analysis pre- and post-virotherapy. 
2.35 
Evaluate transcription of interferon mediated genes (protein kinase R, the death 
receptor-TRAIL, 2’-5’ oligoadenylate/RNAse L proteins, heat shock proteins 
[Hsp 60/70/90], major histocompatibility class antigens and IRF-7). 
2.36 
Assess presence of VSV in tumor and normal tissues subsequent to 
administration of IV VSV-IFNβ-NIS. 
 
 

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3.0 
Patient Eligibility 
3.1 
Inclusion criteria 
3.11 
Age ≥18 years. 
3.12 
Measurable Stage IVA, Stage IVB (with or without measurable disease) or 
recurrent (with or without measurable disease) endometrial carcinoma. 
NOTE: Histologic confirmation of the original primary tumor is required. 
Patients with the following histologic epithelial cell types are eligible: 
Endometrioid adenocarcinoma, serous adenocarcinoma, undifferentiated 
carcinoma, clear cell adenocarcinoma, mixed epithelial carcinoma, 
carcinosarcoma, adenocarcinoma not otherwise specified (NOS). 
NOTE: Measurable disease is defined by RECIST (version 1.1)  
(see Section 11.0). 
3.13 
Group A only: Largest tumor diameter ≤5cm.  
NOTE: Group B patients have no maximum tumor size. 
3.14 
The following laboratory values obtained ≤15 days prior to registration: 
• 
Absolute Neutrophil Count (ANC) ≥1500/μL 
• 
Platelet Count (PLT) ≥100,000/μL 
• 
Hemoglobin ≥10 g/dL 
• 
Creatinine ≤2.0 mg/dL  
• 
AST and ALT ≤2 x upper limit of normal (ULN) 
NOTE: If baseline liver disease, Child Pugh score not exceeding Class A (see 
Appendix III) 
• 
Total bilirubin ≤1.5 x ULN 
• 
INR/PT, aPTT ≤1.4 x ULN  
unless on therapeutic warfarin then INR/PT≤3.5 
3.15 
Ability to provide written informed consent. 
3.16 
Willingness to return to Mayo Clinic in Rochester, Minnesota for follow-up. 
3.17 
Life expectancy ≥12 weeks. 
3.18 
ECOG performance status (PS) 0, 1, or 2 (see Appendix I). 
3.19a 
Willingness to provide mandatory biological specimens for research purposes 
(See Sections 4.0, 14.0, 17.0). 
3.19b Prior therapy:  
3.19b1 
Any number of prior chemotherapy regimens and/or targeted therapies 
and/or prior external beam radiation therapy and/or prior hormonal 
therapy for endometrial cancer are allowed provided the last treatment 
was ≥4 weeks prior to registration. 
3.19b2 
Vaginal brachytherapy may have been administered at any time prior 
to registration. 
 
3.2 
Exclusion criteria 
3.21 
Availability of and patient acceptance of curative therapy.  
3.22 
Active infection requiring treatment, including any active viral infection, ≤5 days 
prior to registration. 

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3.23 
Active or latent tuberculosis or hepatitis. 
3.24 
Known untreated or symptomatic brain metastases. 
3.25 
Any of the following prior therapies: 
• 
Chemotherapy <4 weeks prior to registration 
• 
Targeted biologic therapy<4 weeks prior to registration 
• 
Immunotherapy <4 weeks prior to registration 
• 
Any viral or gene therapy prior to registration 
• 
External beam radiotherapy <4 weeks prior to registration 
NOTE: Vaginal brachytherapy may be performed at any time prior to 
registration 
3.26 
New York Heart Association classification III or IV, known symptomatic 
coronary artery disease, or symptoms of coronary artery disease on systems 
review, or uncontrolled current cardiac arrhythmias (atrial fibrillation or 
supraventricular tachycardia(SVT)) (see Appendix II). 
3.27 
Active CNS disorder or seizure disorder or known CNS disease or neurologic 
symptomatology. 
3.28 
HIV positive test result or other immunodeficiency or immunosuppression. 
3.29a 
History of hepatitis B or C or chronic hepatitis. 
3.29b Other concurrent chemotherapy, immunotherapy, radiotherapy, or any ancillary 
therapy considered investigational (used for a non-FDA approved indication and 
in the context of a research investigation). 
3.29c 
Treatment with oral/systemic corticosteroids, with the exception of topical or 
inhaled steroids. 
3.29d Exposure to household contacts ≤15 months old or household contact with 
known immunodeficiency.  
3.29e  Any of the following because this study involves an investigational agent whose 
genotoxic, mutagenic and teratogenic effects on the developing fetus and 
newborn are unknown: 
• 
Pregnant persons or persons of reproductive ability who are unwilling to use 
effective contraception 
• 
Nursing persons 
3.29f 
Any other pathology or condition that the principal investigator deems to 
negatively impact treatment safety. 
3.29g Any immunotherapy-related adverse events CTCAE >Grade 1 at the time of 
registration.  
3.29h Receipt of a live virus vaccine ≤2 months prior to registration. 
 
 

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4.0 
Test Schedule(s) 
4.1 
Endometrial Cancer Test Schedule for Both Groups 
 
Pre-treatment 
Post-therapy  
(relative to VSV-hIFNβ-NIS administration) 
Observation 
Tests and Procedures1 
≤29 days 
prior to 
Reg 
≤15 days 
prior to 
Reg 
After 
Reg/Prior 
to Tx2 
Day 
1 
Day 
2 
Day 
3 
Day 
4 
Day 
5 
Day 
8 
Day 
10 
Day 
15 
Day 29 
6 weeks 
post 
infusion 
3 mos 
post 
infusion 
At 6, 9, 12 
months3 
Window 
 
 
 
 
 
±1 
±1 
±1 
±1 
±1 -3 to +1 
±3 
±7 days ±7 days 
±14 days 
History and exam, wt, PS 
X 
 
 
X 
X 
 
 
X 
 
 
X 
X 
X 
X 
X 
Pregnancy test 
 
 
X4 
 
 
 
 
 
 
 
 
 
 
 
 
HIV by EIA, Hepatitis panel, Quantiferon 
(TB)R 
XR 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Adverse event assessment, vital signs5 
 
X6 
X7 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
CBC with diff: WBC, ANC, Hgb, PLT 
 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
 
 
 
INR/PT aPTT, fibrinogen 
 
X 
XR 
XR 
 
 
 
 
XR XR 
 
XR 
 
 
 
AST/ALT, Alk Phos, Na, K, T. Bili, Cr, Ca, 
uric acid, phosphorus 
 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
 
 
 
C-reactive protein (CRP)8 
 
 
X 
 
 
 
 
 
 
 
 
 
 
 
 
CA-125 
 
 
XR 
 
 
 
 
 
 
 
XR 
XR 
X 
X 
X 
ECG 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
CT chest/abdomen/pelvis9 
X 
 
 
 
 
 
 
 
 
 
 
X 
 
X 
X 
 
1 Any additional tests and procedures should be performed as needed for clinical care at treating physician’s discretion. 
2 Clinical blood draws for pre-treatment may be done the evening prior due to blood volume required. 
3 Patients will be observed every 3 months for one year after VSV-hIFNβ-NIS administration or until starting another treatment or progression, whichever is 
shorter. Patients will then move to event monitoring. (See Section 13.2) 
4 Only for persons capable of becoming pregnant (NOTE: More than 95% of the patients enrolling in this trial will not be capable of becoming pregnant) 
5 Collect temperature, pulse, blood pressure, and respiratory rate 
6 AE assessment for eligibility is documented on the eligibility checklist. 
7 Baseline AE assessment should be done after registration and prior to treatment. AE assessment at “≤14 days prior to Reg” is solely for eligibility (refer to 
Section 18.5) 
8 Check baseline CRP prior to treatment Day 1; thereafter may be used clinically to monitor for CRS. 
9 CT chest/abdomen/pelvis will be done for baseline eligibility determination; to assess for tumor response at 6 weeks post IV VSV-hIFNβ-NIS infusion; and 
every 3 months until progression of cancer. 

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Pre-treatment 
Post-therapy  
(relative to VSV-hIFNβ-NIS administration) 
Observation 
Tests and Procedures1 
≤29 days 
prior to 
Reg 
≤15 days 
prior to 
Reg 
After 
Reg/Prior 
to Tx2 
Day 
1 
Day 
2 
Day 
3 
Day 
4 
Day 
5 
Day 
8 
Day 
10 
Day 
15 
Day 29 
6 weeks 
post 
infusion 
3 mos 
post 
infusion 
At 6, 9, 12 
months3 
Window 
 
 
 
 
 
±1 
±1 
±1 
±1 
±1 -3 to +1 
±3 
±7 days ±7 days 
±14 days 
Research tissue submission: Tumor biopsy  
(or archived fresh frozen surgical tissue for 
pretreatment only)R 
 
 
XR 
 
 
 
 
 
 
 
 
XR10 
 
XR 
 
Optional NIS imaging11,R 
 
 
XR 
 
XR 
 
 
 
 
XR 
 
 
 
 
 
Biopsy of NIS positive area12,R 
 
 
 
 
 
XR 
 
 
XR 
 
 
 
 
 
 
Viral shedding: mouth rinse, buccal swab, 
and urine collectionR,13 
 
 
XR 
XR XR XR 
 
 
XR XR 
XR 
XR,14 
 
 
 
Research bloodR 
 
 
X15R 
XR XR XR 
 
 
XR XR 
XR 
XR16 
XR 
XR 
XR 
Cycle = 29 days (one cycle) 
R Tests done for research purposes only. 
 
 
 
10 Tumor biopsy will be done on Day 29. Image-guided biopsy may be done under ultrasound-guidance or CT-guidance. 
11 Optional NIS imaging (if available): Up to a total of four (4) scans may be done on each patient. The timing of the scans will be at baseline, Day 2 after VSV-
hIFNβ-NIS infusion, and between Days 7-10 (scan at Days 7-10 only if previous imaging data is positive). Additional scans are at investigator’s discretion. 
12 One optional biopsy of NIS positive area may be done after any NIS imaging if patient agrees. 
13 See Section 14.0 for collection times. 
14 These studies will only be performed if there is evidence of increasing viral proliferation at Day 15. If at Day 29, there is 10-fold increase in VSV-hIFNβ-
NIS/mcg of RNA, repeat tests will be performed within 2 days to confirm the result. Monthly testing will be done thereafter until resolution to baseline. 
15 Pre-treatment research blood (Day 1) to be collected with other samples. 
16 These studies will only be performed if there is evidence of increasing viral proliferation at Day 15. If at Day 29, there is 10-fold increase in VSV-hIFNβ-
NIS/mcg of RNA, repeat tests will be performed within 2 days to confirm the result. Monthly testing will be done thereafter until resolution to baseline. 

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4.2 
Event Monitoring 
CRF 
Event Monitoring Phase* 
q. 
3 mos 
until PD 
At PD 
After PD 
q. 6 mos. 
Death 
New Primary 
Event Monitoring 
X 
X 
X 
X 
At each occurrence 
*If a patient is still alive 5 years after registration, no further follow-up is required. 
5.0 
Grouping Factors 
5.1 
Group: A (VSV-hIFNβ-NIS alone) vs. B (VSV-hIFNβ-NIS with ruxolitinib) 
6.0 
Registration/Randomization Procedures 
6.1 
Phase I Registration 
Prior to discussing protocol entry with the patient, contact the Mayo Clinic 
Research Site Management Office (
) to ensure that a place on 
the protocol is open to the patient.  
6.11 
Registration Procedures 
6.111 To register a patient, fax (
) a completed eligibility checklist 
to the Mayo Clinic Research Site Management Office between 8 a.m. 
and 5 p.m. central time Monday through Friday. 
6.112 MCR only: Patients will be admitted to Mayo Clinic Hospital –Methodist 
Campus in the morning prior to virus administration. The clinical 
research associate (CRA) must inquire with the Medical Oncology unit 
regarding bed availability prior to registering the patient. 
6.2 
All Patients 
6.21 
Correlative Research 
A mandatory correlative research component is part of this study; the patient will 
be automatically registered onto this component (see Sections 3.18, 14.0 and 
17.0). 
6.22 
Registration Staff verification 
Prior to accepting the registration, registration/randomization application will 
verify the following: 
• 
IRB approval at the registering institution 
• 
Patient eligibility 
• 
Existence of a signed consent form 
• 
Existence of a signed authorization for use and disclosure of protected health 
information  
6.23 
IRB Documentation 
Documentation of IRB approval must be on file in the Research Site 
Management Office before an investigator may register any patients.  

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In addition to submitting initial IRB approval documents, ongoing IRB approval 
documentation must be on file (no less than annually) at the Research Site 
Management Office (
 If the necessary documentation is not 
submitted in advance of attempting patient registration, the registration will not 
be accepted and the patient may not be enrolled in the protocol until the situation 
is resolved. 
When the study has been permanently closed to patient enrollment, submission of 
annual IRB approvals to the Research Site Management Office is no longer 
necessary. 
6.24 
Permissions 
At the time of registration, the following will be recorded: 
• 
Patient has/has not given permission to store and use his/her sample(s) for 
future research of cancer at Mayo Clinic. 
• 
Patient has/has not given permission to store and use his/her sample(s) for 
future research to learn, prevent, or treat other health problems at Mayo 
Clinic. 
• 
Patient has/has not given permission for MCCC to give his/her sample(s) to 
researchers at other institutions. 
6.25 
Treatment location 
Treatment on this protocol must commence at Mayo Clinic in Rochester under 
the supervision of a health care professional in the Cancer Center. 
6.26 
Treatment start 
Treatment cannot begin prior to registration and must begin ≤14 days after 
registration. 
 
6.27 
Pretreatment tests/procedures 
Pretreatment tests/procedures (see Section 4.0) must be completed within the 
guidelines specified on the test schedule. 
 
6.28 
Baseline symptoms 
All required baseline symptoms (see Section 10.6) must be documented and 
graded. 
 
.

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7.0 
Protocol Treatment 
This study consists of a single dose escalation scheme to determine the maximum tolerated dose 
(MTD) of VSV-hIFN-NIS when given to patients with metastatic and/or recurrent EC. 
NOTE: Patient must stay within 30 miles of Rochester, MN, for 10-15 days after infusion. 
7.1 
Treatment Schedule 
7.11 
Pre- treatment for VSV-hIFNβ-NIS for all groups 
All patients will receive the following medications prophylactically to mitigate 
fever, hypotension, and nausea. 
Agent 
Dose 
Route 
Day 
Acetaminophen 
650 mg 
PO 
1  
15 minutes prior to infusion  
Diphenhydramine 
25 mg 
IV or PO 
1  
15 minutes prior to infusion  
Ondansetron 
8 mg 
IV or PO 
1  
15 minutes prior to infusion  
Naproxen 
500 mg 
PO 
1 
15 minutes prior to infusion 
7.12 
Group A VSV-hIFNβ-NIS alone 
Agent 
Dose 
Route 
Day 
VSV-hIFNβ-NIS 
As assigned by 
Registration Office* 
IV 
1 
*See Section 7.2 
7.13 
Group B VSV-hIFNβ-NIS with ruxolitinib 
Agent 
Dose 
Route 
Day 
VSV-hIFNβ-NIS 
As assigned by the Registration 
Office 
IV 
1 
Ruxolitinib 
15mg twice per day* 
Oral 
-3 to 9 
*Patients with creatinine clearance between 30-50 ml/mm will start at reduced 
dose of 10mg twice daily  
Note: Ruxolitinib is given for a total of 12 days beginning 3 days prior to virus 
administration. (Patients will continue their own prescription while in the hospital.) 
7.14 
Virus administration (Group A and B) 
The virus will be administered by slow intravenous (IV) infusion (60 minutes) in 
100ml of normal saline with 1% human serum albumin (HSA) and concurrently 
with 1 liter (1000ml) of 0.9% normal saline, under close observation in the 
Medical Oncology Unit, Mayo Clinic Hospital – Methodist Campus.  
After the infusion, the tubing should be flushed using the vascular access protocol. 
The duration of IV infusion can be increased to 90 minutes if an infusion reaction 
is observed. The day of virus administration will be defined as Day 1 of therapy. 

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As a precaution, all patients will remain in the Medical Oncology Unit, Mayo 
Clinic Hospital – Methodist Campus for up to 48 hours after VSV administration. 
Additionally, daily assessments and laboratory tests (CBC, electrolytes, liver 
enzymes, and coagulation panel) will be performed for the first 5 days after VSV 
administration to assess for liver toxicity and tumor lysis syndrome (TLS). 
Because of this requirement, patients will need to stay in Rochester, Minnesota 
for the first 5 days after VSV administration (patient reimbursement for meals 
and lodging is available).  
7.15 
Ruxolitinib (Jakafi®) (Group B only) 
Ruxolitinib is an oral agent that will be given as 15 mg twice per day by mouth 
for 12 days, starting three days before virus infusion. Patients with renal 
impairment at baseline (creatinine clearance 30-50ml) will be started at 10 mg 
twice per day. 
7.16 
Post- treatment for both groups 
All patients will receive IV normal saline for dehydration prophylaxis starting 
2 hours after VSV infusion is complete. 
Agent 
Dose 
Route 
Day 
Normal saline (NS) 
100ml/hour* 
IV 
1  
post infusion 
*Fluid infusion rate may be adjusted per clinical indication 
7.17 
Reaction to the intravenously administered virus 
Patients will be closely monitored in the hospital during infusion of the virus. 
Patients who develop febrile or allergic responses to the infusion will be treated 
with acetaminophen, a cooling blanket, and/or naproxen sodium. For rigors, 
fentanyl may be administered. Symptoms suggestive of anaphylaxis such as 
dyspnea, itching, dizziness or symptomatic hypotension will result in the abrupt 
cessation of the viral infusion (if necessary) and activation of the Rapid Response 
Team (RRT). Administration of all supportive care medications will be tracked in 
the medical record.  
Infusion related reaction (IRR) and symptoms secondary to viremia (this is NOT 
sepsis) could occur 4-6 hours after infusion.  
See Section 9.2 for supportive care guidelines. 
7.18 
Monitoring of viral spread 
The in vivo distribution of VSV-hIFNβ-NIS infected cells and the kinetics of 
virus spread and elimination will be monitored by whole body NIS imaging and 
by serial measurements of viral RNA in mononuclear cells derived from blood, 
saliva and urine (Viral N-gene RNA copy number/μg RNA). 
7.19a 
Optional NIS imaging (if available) 
7.19a1 
Timing of imaging 
Imaging for NIS may be performed at baseline and Day 2 after VSV-
hIFNβ-NIS infusion. Whole-body images will be obtained. If isotope 
uptake is observed in organs other than the thyroid, stomach and 
salivary glands, further imaging may be performed between Days 7-10 
and on Day 15 if needed to document elimination of the virus and virus 

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infected cells. Biopsy will also be taken from accessible NIS image 
positive tumors with patient consent. 
7.19a2 
Planar and SPECT/CT image parameters – No longer in use 
Approximately 20 mCi (±10%) 99mTc pertechnetate will be 
administered intravenously. After a delay of about 15 minutes, whole 
body planar images will be acquired. Subsequently, SPECT/CT 
imaging over two bed positions will be acquired. For most patients, 
this imaging will entail coverage of the thorax, abdomen, pelvis, and 
proximal thighs. This scan range could be adjusted based on any 
comparison imaging acquired before SPECT/CT (i.e., PET/CT, whole 
body low dose CT, radiographic skeletal survey, MRI, etc.) or the 1 
hour planar images demonstrating clinically significant lesions outside 
of the projected scan range. 
7.19a3 
TFB-PET image parameters 
18F-TFB will be prepared and administered per protocol “F-18 TFB-
PET studies in cancer patients undergoing NIS-containing viral 
therapies” (IND 137182). 
Patient preparation: no fasting is required. Patients may be maintained 
on a low iodine diet, stop thyroid-suppressive medications or thyroid 
supplementation, and avoid iodine-containing substances per clinical 
standard of care. 
PET/CT protocol: Scan coverage will extend from upper thighs to the 
base of the skull, starting from skull. At a minimum, 3 minutes per bed 
position will be used. In certain circumstances, coverage may be 
extended to the toes. Low dose CT will be acquired over the same 
range for attenuation correction and anatomic localization. 
7.19a4 
Biopsy of NIS image positive area 
Biopsymay be taken from accessible NIS image-positive tumors after 
any post-treatment NIS imaging. Image-guided biopsy can be 
performed under ultrasound-guidance or CT-guidance.  
7.19b Tumor Biopsy 
Image-guided biopsy of accessible tumor may be performed on Day 29 to assess 
for presence of virus, tumor viability, and immunologic response unless 
circumstances preclude biopsy. 
Attempts will be made to avoid biopsying RECIST target lesions when feasible. 
7.19c 
VSV infection 
Human infection with VSV is uncommon and often asymptomatic. Vesicles on 
the lips and in the mouth or nose occur in about 25% of infected patients and 
have to be distinguished from lesions due to herpes viruses. A swab will be sent 
for herpes virus detection by PCR (Clinical Microbiology laboratory) and blister 
fluid sent for VSV detection by rescue (on Vero cells) and PCR assay. Common 
symptoms include a combination of the following: headache, retroorbital pain, 
fatigue, general malaise, pharyngitis, myalgia, nausea, vomiting and diarrhea.  
The disease is self-limiting in normal adults, treatment will be implemented 
based on clinical judgement if the symptoms (including temperature >38.5°C) 
persist for as long as 6 days from onset or if there are symptoms suggestive of 

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encephalitis, and earlier at the treating physician’s discretion.  
Although there are no clinical trials to support its use, laboratory studies suggest 
that ribavirin (10 mg/kg/day in 4 divided doses orally) can effectively block VSV 
replication. Administration of this medication will be tracked and a participant 
with persistent illness as described above will be classified as having experienced 
a DLT (see Section 7.5). 
Therapy for VSV-hIFNβ-NIS Persistence 
Agent 
Dose 
Route 
Reaction 
Ribavirin 
10 mg/kg/day  
(in 4 divided doses) 
PO 
Persistent VSV- lesions 
Or 
Suspected encephalitis 
Or 
Persistent viremia  
7.19d  Outpatient Hydration Guidelines 
1-2L/day IV normal saline (NS) in Infusion Therapy Center if any of the 
following: 
• Any 1 point increase in AE grade of nausea, vomiting, dehydration, or 
clinically significant electrolyte laboratory values 
• Treating provider discretion 
• If >2L/day IV normal saline required, patient should be hospitalized for IV 
hydration and further management of dehydration. 
7.2 
Dose escalation (Group A and B) 
Three patients will initially be treated at each dose level. The dose of each subsequent 
dose level will be determined by the adverse event evaluations. The starting dose will be  
5 x 109 TCID50 injected intravenously. Between the first and second patient enrolled on 
the first 2 dosing cohorts, there will be a 3 week staggering period managed by the 
enrolling clinical research associate.  
Each group enrolls independently of the other group as the group receiving ruxolitinib 
(Group B) may be able to tolerate a higher dose than Group A. 
As of Amendment 4, 2/6 patients at dose level 2 in Group A have experienced DLT. 
Under the new DLT criteria, dose level 2 for Group A will be re-evaluated with new 
patients. Patients enrolled prior to the new DLT criteria will not be utilized to evaluate 
the MTD.  
Group B will begin enrollment at dose level 1. 
7.21 
Dose escalation table for VSV-hIFNβ-NIS 
Level 
Dose (TCID50) 
-2 
5 × 108 
-1 
1.7 × 109 
1* 
5× 109 
2 
1.7 × 1010 
3 
5 × 1010 
4 
1.7 × 1011 
5 
3.5 × 1011 
*starting dose 

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7.22 
Dose Escalation Rules 
7.221 
DLT is not seen in any of the 3 patients at a given dose level, then 
3 additional patients will be treated at the next dose level. 
7.222 
If DLT is seen in 1 of 3 patients treated at a given dose level, 
3 additional patients will be entered, one by one, at the same dose level. 
7.223 
If no additional DLT is observed at that dose level, then 3 additional 
patients will be treated at the next dose level. 
7.224 
If two or more subjects out of six experiences DLT at any dose level, 
MTD will have been exceeded. 
7.225 
If MTD is exceeded at any dose level, the cohort at the next lower dose 
level will be expanded one by one as needed to reach a total of 
6 participants. 
7.226  
MTD will be defined as the highest dose at which no more than one out 
of six participants experiences DLT. 
7.227  
If no DLT is observed at the highest dose level (i.e., 3.5 × 1011), 
6 additional patients will be enrolled at this dose level and it will be 
defined as the maximum feasible dose. 
7.3 
Dose De-escalation 
7.31 
If two or more patients experience DLT at dose level 1, patients will be entered at 
a lower dose of 1.7 × 109 (see Table in Section 7.21). If two or more patients 
experience DLT at this dose, further patients will be accrued at the -2 dose level 
of 5× 10 8 TCID50. 
7.32 
Toxicity Observation Period: The first 4 weeks after therapy will be considered 
the toxicity observation period, where any DLTs observed in this timeframe will 
determine whether or not accrual can continue to the next dose level. 
7.33 
If a patient fails to complete the initial course of therapy (i.e. registers, but does 
not receive therapy or lost to follow-up during first 4 weeks), the patient will be 
regarded as non-evaluable and an additional patient will be treated at the current 
dose level. For these instances, a specific notation will be made for review by the 
Cancer Center Data Safety Monitoring Board (DSMB). If more than one 
participant must be replaced at a dose level for reasons other than toxicity, the 
reasons will be reported to the FDA and the trial will be voluntarily halted 
pending comments by the FDA review team. 
7.4 
Anticipated toxicity 
7.41 
An acute febrile reaction to the intravenously administered virus may occur. 
Ancillary support is described in Sections 7.12 and 9.2. 
7.42 
A flu-like illness characterized by fever, headache, myalgia, malaise and 
occasionally vesicular lesions around the mouth can occur. Ancillary support is 
described in Sections 7.17 and 9.2. 
7.43 
Liver toxicity with increase in ALT and/or AST, and cytopenia - especially 
lymphopenia - can occur. 
7.44 
Nausea, vomiting, and/or diarrhea with sequelae of dehydration and electrolyte 
imbalances have occurred. 
7.45 
Ruxolitinib is known to cause thrombocytopenia, anemia, and neutropenia. 
7.46 
Patients will be asked to allow supportive care measures including dialysis, if 
needed. 

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7.47 
Patients will be asked to allow autopsy if they die after receiving the vaccine to 
facilitate further research into the vaccine and how it works. 
7.5 
Dose Limiting Toxicities 
7.51 
Dose limiting toxicity (DLT) will be defined as any of the following with 
attribution of at least possibly related to study treatment during the first 28 days: 
Definitions of Dose Limiting Toxicity (DLT) 
CTCAE SOC 
Adverse Event 
Criteria 
Investigations 
Creatinine increased 
Grade 3 (Serum creatinine >3 x upper limit of 
normal) 
Investigations 
Neutrophil count decreased 
Grade 4 for seven (7) days or more (<500/mm3) 
or  
Grade 4 ANC (<500/mm3) with bacterial 
infection 
Investigations 
Platelet count decreased 
Grade 3 for seven (7) days or more 
(<50,000/mm3) 
Grade 4 PLT (<25,000/mm3) 
Immune system  
Allergic reaction (only if not 
associated with viremia -see 
Cytokine release syndrome below) 
≥Grade 2  
disorders 
Autoimmune disorder 
≥Grade 2 with the exception of vitiligo 
Immune system 
disorders 
Cytokine release syndrome** 
Grade ≥3 allergic reactions related to the study 
infusion 
Grade 3 CRS** that does not resolve to 
≤Grade 2 within seven (7) days 
Grade 4 CRS** that does not decrease to 
Grade 2 within 72 hours 
Grade 5 CRS 
Infections and 
infestations 
Infections and infestations – Other 
specify - Life-threatening VSV 
infection 
A combination of three or more of the following 
symptoms, at least two of which are Grade 4, 
resulting in hospitalization or extension of 
hospitalization:  
Malaise, fever, headache, oral ulcerations, 
nausea, vomiting, muscle aches, weakness 
Infections and 
infestations 
Infections and infestations – Other 
specify - Increasing viremia 
(detection of viral RNA by RT-
PCR in PBL) 
>10 fold increase in the copy number (copies/μg 
RNA) between sequential samples at least 
3 days apart beyond Day 14 
All other events Any other event not listed above or 
in Exceptions below table* 
≥ Grade 3 
*With the exception of lymphopenia or other related events (e.g., anemia, white blood 
cell count decreased), which will not be considered a dose limiting toxicity. 
Grade ≥3 flu-like symptoms, fever, nausea, vomiting, dehydration, diarrhea, headache, 
myalgia, fatigue, ALT increased, or AST increased, will also not be considered as a dose 
limiting toxicity as they are anticipated toxicities of treatment (see Section 7.4) 
UNLESS they do not resolve to Grade ≤2 within 7 days of onset in the setting of 
adequate medical management. 

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Other laboratory values that have no clinical correlate, and resolve to Grade ≤2 within 
7 days with adequate medical management; and asymptomatic lipase or amylase 
Grade 3 not associated with clinical manifestations of pancreatitis will not be considered 
a DLT.  
Tumor flare phenomenon defined as local pain, irritation, or rash localized at sites of 
known or suspected tumor that resolve to Grade ≤2 within 6 days will not be considered 
a DLT.  
Symptomatic thyroid dysfunction which is manageable with adequate treatment and 
resolves to ≤Grade 2 within 6 days will also not be considered a DLT. 
**Grading of cytokine release syndrome (CRS) is per Lee et al. Blood, 2014; 124 (2), 
188-195 – See Section 7.52  and Section 9.3. 
7.52 
CRS Grading 
Per Lee et al., Blood, 2014: 124 (2), 188-195 
(https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4093680/) 
CRS Grade 
Description 
Grade 1 CRS 
Fever, constitutional symptoms 
Grade 2 CRS 
Hypotension: responds to fluids or one low dose pressor 
Hypoxia: responds to <40% O2 
Organ toxicity: CTCAE Grade 2 
Grade 3 CRS 
Hypotension: requires multiple pressors or high dose pressors 
Hypoxia: responds to ≥40% O2 
Organ toxicity: CTCAE Grade 3, Grade 4 transaminitis 
Grade 4 CRS 
Mechanical ventilation 
Organ toxicity: Grade 4 excluding transaminitis 
 
 
 

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8.0 
Dosage Modification Based on Adverse Events 
8.1 
VSV-hIFNβ-NIS Dose Modifications 
Given that this is a Phase I trial, there are no dose modifications for VSV-hIFNβ-NIS in 
this study. Patients will only receive a single dose of the virus. 
8.2 
Ruxolitinib Dose Modifications 
Based on discussions with Principal Investigator (PI), ruxolitinib may be omitted or 
modified. Ruxolitinib at a reduced dose of 10 mg per dose may be given at discretion of 
PI and treating physician. 
If patient starts ruxolitinib at reduced dose due to renal or hepatic impairment, then patient 
should stop ruxolitinib if significant worsening occurs, based on discussion with PI. 
 

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9.0 
Ancillary Therapy 
9.1 
Full Supportive Care 
Patients should receive full supportive care during the study including blood products, 
antibiotics and treatment of concurrent medical conditions and other newly diagnosed 
diseases. All blood products and concomitant medications such as antidiarrheals, 
analgesics, and/or antiemetics received from the first day of study treatment 
administration until 30 days after the final dose will be recorded in the medical records. 
9.2 
Concurrent enrollment in other trials 
Patients enrolled in this study will not be eligible for concurrent enrollment in any other 
study involving an interventional pharmacologic agent (drugs, biologicals, 
immunotherapy, gene therapy), whether for therapeutic intent or symptom control. 
Enrollment in non-therapeutic studies will be allowed. 
9.3 
Supportive/Ancillary Care Related to Virus Administration 
9.31 
Reaction to the intravenously administered virus 
Patients will be closely monitored in the hospital during infusion of the virus and 
for up to 48 hours. Patients who develop febrile or allergic responses to the 
infusion will be treated with acetaminophen, cooling blanket, and naproxen. For 
rigors, fentanyl may be administered. Ondansetron may be given for nausea and 
vomiting prophylaxis and treatment, and normal saline will be administered IV to 
reduce the risk of dehydration and electrolyte imbalance. Additional measures 
may be taken at any time if dehydration is suspected, including outpatient 
hydration, and, if necessary, inpatient hydration. 
Symptoms suggestive of anaphylaxis such as dyspnea, itching, dizziness or 
symptomatic hypotension will result in the abrupt cessation of the viral infusion 
and activation of the Rapid Response Team (RRT) for aggressive supportive 
therapy as clinically indicated which may include fluids, hydrocortisone, or other 
agents deemed clinically appropriate. Administration of all supportive care 
medications will be tracked.  
Suggested Therapy for Acute Infusion Reactions* 
Agent 
Dose 
Route 
Reaction 
Acetaminophen and 
cooling blanket 
650 mg q 4h PRN 
PO 
Acute febrile reaction 
Naproxen 
220 mg q8h PRN 
PO 
Acute febrile reaction 
Fentanyl 
25 mcg 
(max 2 doses) 
IV 
Rigors 
Hydrocortisone 
100 mg loading dose 
then 50 mg q6h 
IV 
Hypotension and 
suspected CRS 
Ondansetron 
8mg q 8h PRN 
IV or PO 
Nausea 
*As clinically indicated 
VSV infection in healthy adults is normally self-limiting, however, treatment will 
be implemented if the symptoms (including temperature ≥38.5° C) persist for as 
long as 6 days, if there are symptoms suggestive of encephalitis, and earlier at the 
treating physician’s discretion. Treatment may include ribavirin (10 mg/kg/day in 

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4 divided doses orally or 20mg/kg/day intravenously). Administration of these 
medications will be tracked and a participant with persistent symptoms or 
encephalitis will be classified as having experienced a DLT (see Section 7.5). 
9.32 
Liver toxicity 
Patients experiencing ≥Grade 3 liver toxicity (AST/ALT ≥5X upper limit of 
normal (ULN), bilirubin ≥3X ULN, or alkaline phosphatase (APPT) ≥2.5X 
ULN) will be admitted to the hospital. Universal precautions will be followed. 
Blood, urine, and saliva will be evaluated using PCR to evaluate for viremia and 
viral shedding, and plasma IFN beta levels. If there is any evidence of 
increasingly high IFN beta levels, or viremia or viral shedding, ribavirin may be 
administered as described in Section 9.2. The patient will be evaluated for tumor 
lysis syndrome with serum K+, Ca++, phosphate, and uric acid levels. If 
evidence of tumor lysis syndrome is present, the patient will be hydrated and 
treated with rasburicase as clinically indicated. Daily monitoring of hematologic 
parameters and serum chemistries will be performed, vital signs and urine output 
will be monitored, and all other clinically indicated parameters will be 
monitored. Transfer to the intensive care unit with full supportive care will be 
performed if clinically indicated. High IFN beta levels can potentially result in 
liver toxicity, and ruxolitinib has been shown to ameliorate it in murine models. 
Ruxolitinib may be given orally at 15 mg for 14 days or at physician discretion. 
9.33 
Persistent viremia 
Patients who demonstrate persistent and increasing viremia or viral shedding (10-
fold increases in VSV-hIFNβ-NIS genome/mcg RNA at 4 weeks) that is 
confirmed on 2 consecutive samples may be treated with ribavirin. 
9.34 
Viral transmission 
VSV is not spread from person to person and there is a very short viremic phase 
with natural infection. However, the patient, nursing staff as well as immediate 
family of the patient will be educated on the potential role of perioral vesicles in 
the transmission of the virus since this is thought to be possible in some 
mammals. 
9.35 
Cytokine Release Syndrome (CRS) and Interferon beta (IFNβ or IFN) toxicity 
Please note for this treatment regimen CRS is generally early onset (within a few 
hours after virus administration) and associated with fevers, hypotension, and 
hypoxemia. The syndrome typically resolves by 48 hours post-dose and is not 
associated with major IL-6 elevation. 
Interferon beta (IFNβ or IFN) toxicity is usually later onset (Days 3-8) associated 
with abnormal liver function tests and thrombocytopenia, with or without 
hypotension and hypoxemia. 
Following administration of VSV-hIFNβ-NIS, a small percentage (<2%) of 
patients are at risk of severe or life-threatening IFNβ toxicity associated with 
TLS. Immediately after administration, VSV-hIFNβ-NIS extravasates into tumor 
and seeds infection, which can rapidly expand, and in a small percentage of 
patients release large amounts of IFNβ that can result in inflammatory lysis of 
infected tumor cells with systemic sequelae [Miller 2014]. The timing and 
duration of CRS, IFNβ toxicity and TLS are depicted in Figure 9.35a, and their 

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respective unique and distinguishing features are noted in Table 9.371. 
Figure 9.35a 
Timing of toxicities of intravenous VSV  
 
Following intravenous delivery, extensive intra-tumoral extravasation and 
subsequent propagation of the VSV-IFNβ-NIS virus infection at sites of tumor 
growth may occasionally lead to very high serum IFNβ levels (above 10,000 
pg/mL), peaking within the first 72 hours after virus infusion and returning to 
baseline within one to two weeks. Sustained high circulating concentrations of 
virus-encoded IFNβ can lead to a potentially fatal IFNβ toxicity syndrome with 
onset 48 to 96 hours after virus infusion. The syndrome is characterized by 
rapidly rising liver enzyme levels and falling platelet counts which may be seen 
in isolation or in association with persistent/worsening malaise, hypotension, 
and/or neurological symptoms such as confusion and aphasia. Untreated, the 
syndrome may progress to multiorgan failure and may be compounded by tumor 
lysis syndrome (TLS) due to synchronous death of virus-infected tumor cells.  
9.36 
CRS 
Suggested supportive care based on Lee et al., 
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4093680/ 
Grade 1 CRS – Vigilant supportive care; assess for infection. Treat fever and 
neutropenia if present.  
Monitor fluid balance.  
Use antipyretics, analgesics as needed. 
Grade 2 CRS without extensive co-morbidities and not older age – vigilant 
supportive care; monitor cardiac and other organ function closely 
Grade 2 CRS with extensive co-morbidities or older age and Grades 3-4 CRS – 
vigilant supportive care 
NOTE: Vigilant supportive care including empiric treatment of concurrent 
bacterial infections and maintenance of adequate hydration and blood pressure 
for every grade. Immunosuppression (corticosteroids) should be used in all 
patients with Grade 3 or 4 CRS and instituted earlier in patients with extensive 
comorbidities or older age. Grades 2-4 organ adverse events are dictated by 
CTCAE v4.0. 
9.37 
IFN toxicity 
Additional monitoring and evaluations are necessary for early detection of 
persistent signs and symptoms of IFNβ toxicity or additional related clinical 
toxicities. Therefore, all enrolled patients are required to be hospitalized 

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overnight on day of infusion and discharged the following day (Day 2) if 
clinically stable. Once outpatient, the patient will be clinically evaluated in 
person on study Days 3, 4, and 5, including safety labs. Table 9.371 outlines the 
specific IFNβ toxicities and their respective grading per NCI CTCAE v5.0. 
Table 9.371 Signs and symptoms of interferon beta toxicity using CTCAE v5.0 
CTCAE Term 
Grade 1 
Grade 2 
Grade 3 
Grade 4 
Grade 5 
Aspartate 
aminotransferase 
(AST) increased 
>ULN – 3.0 x 
ULN if baseline 
was normal; 1.5 – 
3.0 x baseline if 
baseline was 
abnormal 
>3.0 – 5.0 x ULN 
if baseline was 
normal; >3.0 – 
5.0 x baseline if 
baseline was 
abnormal 
>5.0 – 20.0 x 
ULN if baseline 
was normal; >5.0 
– 20.0 x baseline 
if baseline was 
abnormal 
>20.0 x ULN if 
baseline was 
normal; >20.0 x 
baseline if 
baseline was 
abnormal 
- 
Hypotension 
Asymptomatic, 
intervention not 
indicated 
Non-urgent 
medical 
intervention 
indicated 
Medical 
intervention 
indication; 
hospitalization 
indicated 
Life-threatening 
consequences 
and urgent 
intervention 
indicated 
Death 
Platelet count 
decreased 
<LLN – 
75,000/mm3 
<LLN – 75.0 x 
10e9 / L 
<75,000 – 
50,000/mm3 
<75.0 – 50.0 x 
10e9 /L 
<50,000 – 
25,000/mm3; 
<50.0 – 25.0 x 
10e9 / L 
< 25,000/mm3; 
<25.0 x 10e9 / L 
- 
Persistence of vital signs perturbations at any time point from Day 3 onwards, 
particularly hypotension and/or hypoxia (patient requires supplemental oxygen or 
an increase in baseline supplemental oxygen requirement), should trigger 
inpatient observation and initiation (or extension) of ruxolitinib. Investigators 
should notify the Sponsor/Principal Investigator should they have any doubt 
about safety monitoring and need for inpatient management.  
Investigators are alerted to be vigilant for persistent and/or delayed signs and 
symptoms of CRS and/or early signs of tumor lysis syndrome (TLS). Clinical 
evidence of tumor lysis justifies implementation of TLS management. 
Based on pre-clinical data [Zhang 2016] and emerging clinical experience, 
persistent prolonged CRS and/or IFNβ toxicity is indicative of VSV in vivo 
proliferation and substantive viremia. Ruxolitinib has been demonstrated to 
reduce IFNβ-mediated JAK-STAT signaling. Therefore, ruxolitinib treatment 
should be implemented at the trigger events listed in Table 9.372. 
Table 9.372 Algorithm for implementation or extension of ruxolitinib 
treatment 
Day 
Clinical and/or Lab Findings 
Management 
Day 3 
• 
≥ Gr 3 Hypotension1  
OR 
• 
Hypoxia requiring supplemental oxygen 
OR 
• 
New onset ≥ Gr3 or ≥ 5x baseline AST 
elevation 
• 
Admit for inpatient 
observation 
• 
Aggressive supportive care 
• 
Initiate treatment with oral 
allopurinol for TLS 
prophylaxis2 
• 
≥ Gr 3 Hypotension1  
OR 
• 
Hypoxia requiring supplemental oxygen 
AND 
• 
As above  
• 
Initiate ruxolitinib 15mg PO 
BID 

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Day 
Clinical and/or Lab Findings 
Management 
• 
New onset ≥ Gr3 or ≥ 5x baseline AST 
elevation 
OR 
• 
New onset neurologic symptoms 
Day 4  
 
or  
 
Day 5 
• 
≥ Gr 3 Hypotension1  
OR 
• 
New onset ≥ Gr3 or ≥ 5x baseline AST 
elevation  
OR 
• 
Hypoxia requiring supplemental oxygen 
OR 
• 
If already inpatient and signs and 
symptoms do not improve (e.g., 
persistent or worsening clinical condition 
with new signs/ symptoms such as ≥ Gr 
3 thrombocytopenia, neurological 
symptoms, or TLS) 
OR 
• 
Rising AST elevation 
• 
Admit for inpatient 
observation 
• 
Aggressive supportive care 
• 
If not already initiated, begin 
treatment with oral allopurinol 
for TLS prophylaxis2 
• 
Initiate ruxolitinib 15mg PO 
BID 
• 
If already on ruxolitinib, 
consider increasing to 25mg 
BID 
• 
Rasburicase, if indicated 
• 
Consider steroids 
1 per CTCAE v5. 
2 Standard dose of allopurinol should be initiated for TLS prophylaxis; adjustments for 
renal failure or other causes are permitted. 
If conditions above are met, the patient will be admitted for inpatient observation, 
and symptomatic treatment will be initiated per institutional guidelines, along 
with ruxolitinib administration. If additional clinical findings are present 
suggestive of IFNβ toxicity, such as neurological symptoms or other clinical 
conditions, additional supportive treatment measures should be considered, such 
as corticosteroids. It is up to physician discretion whether treatment should be 
initiated earlier, such as if the patient has worsening clinical condition on Day 2; 
this change should first be discussed with the Principal Investigator. 
9.38 
Neutropenia 
Patients experiencing neutropenia (ANC <500/μL) may be treated with 
levofloxacin (500mg orally daily) or with cefdinir (300 mg orally every 12 hours 
if patient has allergy to levofloxacin) until neutropenia resolves.  
 
 
 

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10.0 
Adverse Event (AE) Reporting and Monitoring 
The site principal investigator is responsible for reporting any/all serious adverse events to the 
sponsor as described within the protocol, regardless of attribution to study agent or treatment 
procedure. 
The sponsor/sponsor-investigator is responsible for notifying FDA and all participating 
investigators in a written safety report of any of the following: 
• 
Any suspected adverse reaction that is both serious and unexpected. 
• 
Any findings from laboratory animal or in vitro testing that suggest a significant risk for 
human subjects, including reports of mutagenicity, teratogenicity, or carcinogenicity. 
• 
Any findings from epidemiological studies, pooled analysis of multiple studies, or 
clinical studies, whether or not conducted under an IND and whether or not conducted by 
the sponsor, that suggest a significant risk in humans exposed to the drug 
• 
Any clinically important increase in the rate of a serious suspected adverse reaction over 
the rate stated in the protocol or Investigator’s Brochure (IB). 
Summary of SAE Reporting for this study  
(please read entire section for specific instructions): 
WHO: 
WHAT form: 
WHERE to send: 
All 
sites 
Pregnancy Reporting 
 
Mayo Sites – attach to MCCC 
Electronic SAE Reporting Form and 
copy Vyriad Pharmacovigilance at 
 
Mayo 
Clinic 
Sites 
Mayo Clinic Cancer Center SAE Reporting Form:  
Will automatically be sent to 
and copy Vyriad 
Pharmacovigilance at  
 
Mayo 
Clinic 
Sites 
Mayo Clinic Institutional Biosafety Reporting: 
 
Mayo Sites – Attach to MCCC 
Electronic SAE Reporting Form 
*Vyriad has an agreement with Mayo Clinic to receive all AEs for VSV-hIFNb-NIS 
Definitions 
Adverse Event 
Any untoward medical occurrence associated with the use of a drug in humans, whether or 
not considered drug related.  
Suspected Adverse Reaction 
Any adverse event for which there is a reasonable possibility that the drug caused the adverse 
event. 
Expedited Reporting 
Events reported to sponsor within 24 hours, 5 days or 10 days of study team becoming aware 
of the event. 

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Routine Reporting 
Events reported to sponsor via case report forms 
Events of Interest 
Events that would not typically be considered to meet the criteria for expedited reporting, but 
that for a specific protocol are being reported via expedited means in order to facilitate the 
review of safety data (may be requested by the FDA or the sponsor).  
Unanticipated Adverse Device Event (UADE) 
Any serious adverse effect on health or safety or any life-threatening problem or death caused 
by, or associated with, a device, if that effect, problem, or death was not previously identified 
in nature, severity, or degree of incidence in the investigational plan or application (including 
a supplementary plan or application), or any other unanticipated serious problem associated 
with a device that relates to the rights, safety, or welfare of subjects 
10.1 
Adverse Event Characteristics 
CTCAE term (AE description) and grade: The descriptions and grading scales found 
in the revised NCI Common Terminology Criteria for Adverse Events (CTCAE) version 
4.0 will be utilized for AE reporting. All appropriate treatment areas should have access 
to a copy of the CTCAE version 4.0. A copy of the CTCAE version 4.0 can be 
downloaded from the CTEP web site: 
 
a. Identify the grade and severity of the event using the CTCAE version 4.0. 
b. Determine whether the event is expected or unexpected (see Section 10.2). 
c. Determine if the adverse event is related to the study intervention (agent, treatment or 
procedure) (see Section 10.3). 
d. Determine whether the event must be reported as an expedited report. If yes, 
determine the timeframe/mechanism (see Section 10.4).  
e. Determine if other reporting is required (see Section 10.5). 
f. Note: All AEs reported via expedited mechanisms must also be reported via the routine 
data reporting mechanisms defined by the protocol (see Sections 10.6 and 18.0). 
NOTE: A severe AE is NOT the same as a serious AE, which is defined in Section 10.4. 
10.2 
Expected vs. Unexpected Events  
Expected events - are those described within the Section 15.0 of the protocol, the study 
specific consent form, package insert (if applicable), and/or the investigator brochure, (if 
an investigator brochure is not required, otherwise described in the general 
investigational plan).  
Unexpected adverse events or suspected adverse reactions are those not listed in Section 
15.0 of the protocol, the study specific consent form, package insert (if applicable), or in 
the investigator brochure (or are not listed at the specificity or severity that has been 
observed); if an investigator brochure is not required or available, is not consistent with 
the risk information described in the general investigational plan. 
Unexpected also refers to adverse events or suspected adverse reactions that are 
mentioned in the investigator brochure as occurring with a class of drugs but have not 
been observed with the drug under investigation. 
An investigational agent/intervention might exacerbate the expected AEs associated with a 
commercial agent. Therefore, if an expected AE (for the commercial agent) occurs with a 
higher degree of severity or specificity, expedited reporting is required. 

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NOTE: *The consent form may contain study specific information at the discretion of the 
Principal Investigator; it is possible that this information may NOT be included in the 
protocol or the investigator brochure. Refer to protocol or IB for reporting needs. 
10.3 
Attribution to agent(s) or procedure 
When assessing whether an adverse event (AE) is related to a medical agent(s) medical or 
procedure, the following attribution categories are utilized: 
Definite - The AE is clearly related to the agent(s)/procedure. 
Probable - The AE is likely related to the agent(s)/procedure. 
Possible - The AE may be related to the agent(s)/procedure. 
Unlikely - The AE is doubtfully related to the agent(s)/procedure. 
Unrelated - The AE is clearly NOT related to the agent(s)/procedure. 
10.31 EXPECTED Serious Adverse Events: Protocol Specific Exceptions to 
Expedited Reporting 
For this protocol only, the following Adverse Events/Grades are expected to occur 
within this population and do not require Expedited Reporting. These events must still 
be reported via Routine Reporting (see Section 10.6).* 
*Report any clinically important increase in the rate of a serious suspected 
adverse reaction (at your study site) over that which is listed in the protocol or 
investigator brochure as an expedited event.  
*Report an expected event that is greater in severity or specificity than expected 
as an expedited event. 
CTCAE System Organ 
Class (SOC) 
Adverse event/ Symptoms 
CTCAE Grade at which the event 
will not be reported via expedited 
mechanisms1 
Blood and lymphatic 
system disorders 
Anemia 
≤Grade 4 
Investigations 
Lymphocyte count decreased 
≤Grade 4 
Neutrophil count decreased 
≤Grade 4 
 
Platelet count decreased 
≤Grade 4 
White blood cell count 
decreased 
≤Grade 4 
1 These exceptions only apply if the adverse event does not result in hospitalization. If the adverse 
event results in hospitalization, then the standard expedited adverse events reporting 
requirements must be followed.  
Specific protocol exceptions to expedited reporting should be reported expeditiously by 
investigators ONLY if they exceed the expected grade of the event. 
The following hospitalizations are not considered to be SAEs because there is no 
“adverse event” (i.e., there is no untoward medical occurrence) associated with the 
hospitalization: 
● Hospitalizations for respite care 
● Planned hospitalizations required by the protocol 
● Hospitalization planned before informed consent (where the condition requiring the 
hospitalization has not changed post study drug administration) 
● Hospitalization for elective procedures unrelated to the current disease and/or treatment 
on this trial 

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● Hospitalization for administration of study drug or insertion of access for 
administration of study drug 
● Hospitalization for routine maintenance of a device (e.g., battery replacement) that was 
in place before study entry 
● Hospitalization or other serious outcomes for signs and symptoms of progression of the 
cancer. 
10.4 
Expedited Reporting Requirements for IND/IDE Agents  
10.41 Phase 1 and Early Phase 2 Studies: Expedited Reporting Requirements for 
Adverse Events that Occur on Studies under an IND/IDE within 30 Days of 
the Last Administration of the Investigational Agent/Intervention 1, 2 
FDA REPORTING REQUIREMENTS FOR SERIOUS ADVERSE EVENTS (21 CFR Part 312) 
NOTE: Investigators MUST immediately report to the sponsor ANY Serious Adverse Events, whether or not they 
are considered related to the investigational agent(s)/intervention (21 CFR 312.64) 
An adverse event is considered serious if it results in ANY of the following outcomes:  
1) 
Death 
2) 
A life-threatening adverse event  
3) 
An adverse event that results in inpatient hospitalization or prolongation of existing hospitalization for ≥ 24 
hours  
4) 
A persistent or significant incapacity or substantial disruption of the ability to conduct normal life functions  
5) 
A congenital anomaly/birth defect.  
6) 
Important Medical Events (IME) that may not result in death, be life threatening, or require hospitalization 
may be considered serious when, based upon medical judgment, they may jeopardize the patient or subject 
and may require medical or surgical intervention to prevent one of the outcomes listed in this definition. (FDA, 
21 CFR 312.32; ICH E2A and ICH E6). 
ALL SERIOUS adverse events that meet the above criteria MUST be immediately reported to the sponsor within 
the timeframes detailed in the table below.  
Hospitalization 
Grade 1 and Grade 2 Timeframes 
Grade 3-5 
Timeframes 
Resulting in 
Hospitalization 
≥24 hrs 
7 Calendar Days 
24-Hour 3 Calendar 
Days 
Not resulting in 
Hospitalization 
≥24 hrs 
Not required 
Expedited AE reporting timelines are defined as: 
o “24-Hour; 3 Calendar Days” - The AE must initially be reported within 24 hours of learning of the AE, 
followed by a complete expedited report within 3 calendar days of the initial 24-hour report. 
o “7 Calendar Days” - A complete expedited report on the AE must be submitted within 7 calendar days 
of learning of the AE. 
1Serious adverse events that occur more than 30 days after the last administration of investigational 
agent/intervention and have an attribution of possible, probable, or definite require reporting as follows:  
Expedited 24-hour notification followed by complete report within 3 calendar days for: 
• All Grade 3, 4, and Grade 5 AEs 
Expedited 7 calendar day reports for: 
• Grade 2 AEs resulting in hospitalization or prolongation of hospitalization 
2 For studies using PET or SPECT IND agents, the AE reporting period is limited to 10 radioactive half-lives, 
rounded UP to the nearest whole day, after the agent/intervention was last administered. Footnote “1” above 
applies after this reporting period. 
Effective Date: May 5, 2011 
NOTE: Refer to Section 10.31 for exceptions to Expedited Reporting 

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10.42 General reporting instructions 
The Mayo IND Coordinator will assist the sponsor-investigator in the processing 
of expedited adverse events and forwarding of suspected unexpected serious 
adverse reactions (SUSARs) to the FDA and IRB. 
Use Mayo Expedited Event Report form 
 
arm.  
Send a copy to Vyriad Pharmacovigilance at 
  
NOTE: Vyriad has an agreement with Mayo Clinic to receive all SAEs reported 
for VSV-hIFNb-NIS. 
10.43 Biotechnology/Biosafety Reporting 
For studies requiring reporting to NIH Office of Biotechnology Activities (NIH 
OBA), Mayo Clinic Cancer Center Institutions: Use the Adverse Event Reporting 
form located on the Mayo Clinic Biosafety Website 
go to the SAE sub-page. 
Mayo Clinic Cancer Center (MCCC) Institutions: 
Attach copies to the Mayo Clinic Cancer Center Adverse Event Reporting 
System 
which will automatically be sent 
to the following email address: 
This 
email will be managed by the SAE, IND and Safety Reporting Coordinators. 
10.44 Reporting of re-occurring SAEs 
ALL SERIOUS adverse events that meet the criteria outlined in table10.41 
MUST be immediately reported to the sponsor within the timeframes detailed in 
the corresponding table. This reporting includes, but is not limited to SAEs that 
re-occur again after resolution. 
10.5 
Other Required Reporting 
10.51 Unanticipated Problems Involving Risks to Subjects or Others (UPIRTSOS) 
Unanticipated Problems Involving Risks to Subjects or Others (UPIRTSOS) in 
general, include any incident, experience, or outcome that meets all of the 
following criteria: 
1. Unexpected (in terms of nature, severity, or frequency) given (a) the research 
procedures that are described in the protocol-related documents, such as the 
IRB-approved research protocol and informed consent document; and (b) the 
characteristics of the subject population being studied; 
2. Related or possibly related to participation in the research (in this guidance 
document, possibly related means there is a reasonable possibility that the 
incident, experience, or outcome may have been caused by the procedures 
involved in the research); and 
3. Suggests that the research places subjects or others at a greater risk of harm 
(including physical, psychological, economic, or social harm) than was 
previously known or recognized. 

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Some unanticipated problems involve social or economic harm instead of the 
physical or psychological harm associated with adverse events. In other cases, 
unanticipated problems place subjects or others at increased risk of harm, but no 
harm occurs. 
Note: If there is no language in the protocol indicating that pregnancy is not 
considered an adverse experience for this trial, and if the consent form does not 
indicate that subjects should not get pregnant/impregnate others, then any 
pregnancy in a subject/patient or a male patient’s partner (spontaneously 
reported) which occurs during the study or within 120 days of completing the 
study should be reported as a UPIRTSO. 
Mayo Clinic Cancer Center (MCCC) Institutions: 
If the event meets the criteria for IRB submission as a Reportable 
Event/UPIRTSO, provide appropriate documentation to 
via Mayo Clinic Cancer Center Adverse 
Event Reporting System 
which will automatically be sent 
to the following email address. The Mayo Regulatory Affairs Office will review 
and process the submission to the Mayo Clinic IRB. 
10.52 Death 
Note: A death on study requires both routine and expedited reporting 
regardless of causality, unless as noted below. Attribution to treatment or 
other cause must be provided. 
Any death occurring within 30 days of the last dose, regardless of attribution to 
an agent/intervention under an IND/IDE requires expedited reporting within 
24-hours. 
Any death occurring greater than 30 days with an attribution of possible, 
probable, or definite to an agent/intervention under an IND/IDE requires 
expedited reporting within 24-hours. 
Reportable categories of Death  
• 
Death attributable to a CTCAE term. 
• 
Death Neonatal: A disorder characterized by cessation of life during the 
first 28 days of life.  
• 
Death NOS: A cessation of life that cannot be attributed to a CTCAE term 
associated with Grade 5. 
• 
Sudden death NOS: A sudden (defined as instant or within one hour of the 
onset of symptoms) or an unobserved cessation of life that cannot be 
attributed to a CTCAE term associated with Grade 5. 
• 
Death due to progressive disease should be reported as Grade 5 
“Neoplasms benign, malignant and unspecified (including cysts and 
polyps) – Other (Progressive Disease)” under the system organ class 
(SOC) of the same name. Evidence that the death was a manifestation of 
underlying disease (e.g., radiological changes suggesting tumor growth or 
progression: clinical deterioration associated with a disease process) 
should be submitted. 

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Per NIH OBA Appendix M, should a patient die during the study or study 
follow-up, no matter what the cause, the study doctor will ask the patient’s 
family for permission to perform an autopsy. If permission is granted, a copy 
of the autopsy report will be sent to the sponsor after all identifying 
information has been removed. An autopsy will help the researchers learn 
more about the safety and efficacy of the treatment. Patients should advise 
their families about their wishes regarding autopsy. 
10.53 Secondary Malignancy 
• 
A secondary malignancy is a cancer caused by treatment for a previous 
malignancy (e.g., treatment with investigational agent/intervention, radiation 
or chemotherapy). A secondary malignancy is not considered a metastasis of 
the initial neoplasm. 
• 
All secondary malignancies that occur following treatment with an agent under 
an IND/IDE will be reported. Three options are available to describe the event: 
o Leukemia secondary to oncology chemotherapy (e.g., Acute Myeloctyic 
Leukemia [AML]) 
o Myelodysplastic syndrome (MDS) 
o Treatment-related secondary malignancy 
• 
Any malignancy possibly related to cancer treatment (including AML/MDS) 
should also be reported via the routine reporting mechanisms outlined in each 
protocol. 
10.54 Second Malignancy 
A second malignancy is one unrelated to the treatment of a prior malignancy (and is 
NOT a metastasis from the initial malignancy). Second malignancies require ONLY 
routine reporting unless otherwise specified. 
10.55 Pregnancy, Fetal Death, and Death Neonatal  
If a female subject (or female partner of a male subject) taking investigational 
product becomes pregnant, the subject taking should notify the Investigator, and 
the pregnant female should be advised to call her healthcare provider 
immediately. The patient should have appropriate follow-up as deemed necessary 
by her physician. If the baby is born with a birth defect or anomaly, a second 
expedited report is required. 
Prior to obtaining private information about a pregnant woman and her infant, the 
investigator must obtain consent from the pregnant woman and the newborn 
infant’s parent or legal guardian before any data collection can occur. A consent 
form will need to be submitted to the IRB for these subjects if a pregnancy 
occurs. If informed consent is not obtained, no information may be collected. 
In cases of fetal death, miscarriage or abortion, the mother is the patient. In cases 
where the child/fetus experiences a serious adverse event other than fetal death, 
the child/fetus is the patient. 
NOTE: When submitting Mayo Expedited Adverse Event Report reports for 
“Pregnancy”, “Pregnancy loss”, or “Neonatal loss”, the potential risk of exposure 
of the fetus to the investigational agent(s) or chemotherapy agent(s) should be 
documented in the “Description of Event” section. Include any available medical 
documentation. Include this form: 

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10.551 
Pregnancy 
Pregnancy should be reported in an expedited manner as Grade 3 
“Pregnancy, puerperium and perinatal conditions - Other 
(pregnancy)” under the Pregnancy, puerperium and perinatal 
conditions SOC. Pregnancy should be followed until the outcome is 
known. 
10.552 
Fetal Death 
Fetal death is defined in CTCAE as “A disorder characterized by death 
in utero; failure of the product of conception to show evidence of 
respiration, heartbeat, or definite movement of a voluntary muscle after 
expulsion from the uterus, without possibility of resuscitation.” 
Any fetal death should be reported expeditiously, as Grade 4 
“Pregnancy, puerperium and perinatal conditions - Other 
(pregnancy loss)” under the Pregnancy, puerperium and perinatal 
conditions SOC. 
10.553 
Death Neonatal 
Neonatal death, defined in CTCAE as “A disorder characterized by 
cessation of life occurring during the first 28 days of life” that is felt by 
the investigator to be at least possibly due to the investigational 
agent/intervention, should be reported expeditiously.  
A neonatal death should be reported expeditiously as Grade 4 
“General disorders and administration - Other (neonatal loss)” 
under the General disorders and administration SOC.  
10.6 
Required Routine Reporting  
10.61 Baseline and Adverse Events Evaluations 
Pretreatment symptoms/conditions to be graded at baseline and adverse events to 
be graded at each evaluation.  
Grading is per CTCAE v4.0 unless alternate grading is indicated in the table below: 
CTCAE  
System/Organ/Class 
(SOC) 
Adverse event/Symptoms 
Baseline 
Each 
evaluation 
Grading scale  
(if not CTCAE) 
Immune system 
disorders 
Allergic reaction (unrelated 
to viremia – see CRS below) 
 
X 
 
Immune system 
Autoimmune disorder 
 
X 
 
disorders 
Cytokine release syndrome 
 
X 
Lee et al. Blood 2014; 124 (2), 
188-195 
https://www.ncbi.nlm.nih.gov/
pmc/articles/PMC4093680/ 
Infections and 
infestations 
Infections and infestations – 
Other: specify – Life-
threatening VSV infection 
 
X 
 
Infections and 
infestations 
Infections and infestations – 
Other specify - Increasing 
viremia  
(detection of viral RNA by 
RT-PCR in PBL) 
 
X 
>10 fold increase in the copy 
number (copies/μg RNA) 
between sequential samples at 
least 3 days apart beyond 
Day 14 

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CTCAE  
System/Organ/Class 
(SOC) 
Adverse event/Symptoms 
Baseline 
Each 
evaluation 
Grading scale  
(if not CTCAE) 
Investigations 
Creatinine increased 
X 
X 
 
 
Neutrophil count decreased  
X 
X 
 
 
Platelet count decreased 
X 
X 
 
 
10.62 Submit via appropriate MCCC Case Report Forms (i.e., paper or electronic, as 
applicable) the following AEs experienced by a patient and not specified in 
Section 10.6: 
10.621 Grade 1 and 2 AEs deemed possibly, probably, or definitely related to the 
study treatment or procedure. 
10.622 Grade 3 and 4 AEs regardless of attribution to the study treatment or 
procedure. 
10.623 Grade 5 AEs (Deaths) 
10.6231 
Any death within 30 days of the patient’s last study 
treatment or procedure regardless of attribution to the study 
treatment or procedure. 
10.6232 
Any death more than 30 days after the patient’s last study 
treatment or procedure that is felt to be at least possibly 
treatment related must also be submitted as a Grade 5 AE, 
with a CTCAE type and attribution assigned. 
10.7 
Late Occurring Adverse Events 
Refer to the instructions in the Forms Packet (or electronic data entry screens, as 
applicable) regarding the submission of late occurring AEs following completion of the 
Active Monitoring Phase (i.e., compliance with Test Schedule in Section 4.0). 
 

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11.0 
Treatment Evaluation. 
11.1 
Schedule of Evaluations 
For the purposes of this study, patients will be evaluated for response at 6 weeks and 
every 3 months thereafter. 
11.2 
Definitions of Measurable and Non-Measurable Disease (RECIST 1.1 criteria): 
11.21 Measurable Disease 
11.211 
A non-nodal lesion is considered measurable if its longest diameter can 
be accurately measured as ≥2.0 cm with chest x-ray, or as ≥1.0 cm 
with CT scan or MRI. 
11.212 
A superficial non-nodal lesion is measurable if its longest diameter is 
≥1.0 cm in diameter as assessed using calipers (e.g. skin nodules) or 
imaging. In the case of skin lesions, documentation by color 
photography, including a ruler to estimate the size of the lesion, is 
recommended.  
11.213 
A malignant lymph node is considered measurable if its short axis is 
>1.5 cm when assessed by CT scan (CT scan slice thickness 
recommended to be no greater than 5 mm). 
NOTE: Tumor lesions in a previously irradiated area are not considered 
measurable disease. 
11.22 Non-Measurable Disease 
11.221 
All other lesions (or sites of disease) are considered non-measurable 
disease, including pathological nodes (those with a short axis ≥1.0 to 
<1.5 cm). Bone lesions, leptomeningeal disease, ascites, 
pleural/pericardial effusions, lymphangitis cutis/pulmonis, 
inflammatory breast disease, and abdominal masses (not followed by 
CT or MRI), are considered as non- measurable as well. 
Note: ‘Cystic lesions’ thought to represent cystic metastases can be considered as 
measurable lesions, if they meet the definition of measurability described above. 
However, if non-cystic lesions are present in the same patient, these are preferred 
for selection as target lesions. In addition, lymph nodes that have a short axis 
<1.0 cm are considered non-pathological (i.e., normal) and should not be 
recorded or followed. 
11.3 
Guidelines for Evaluation of Measurable Disease 
11.31 Measurement Methods: 
• 
All measurements should be recorded in metric notation (i.e., decimal 
fractions of centimeters) using a ruler or calipers. 
• 
The same method of assessment and the same technique must be used to 
characterize each identified and reported lesion at baseline and during follow- 
up. For patients having only lesions measuring at least 1 cm to less than 2 cm 
must use CT imaging for both pre- and post-treatment tumor assessments. 
• 
Imaging-based evaluation is preferred to evaluation by clinical examination 
when both methods have been used at the same evaluation to assess the 
antitumor effect of a treatment. 

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11.32 Acceptable Modalities for Measurable Disease: 
• 
Conventional CT and MRI: This guideline has defined measurability of 
lesions on CT scan based on the assumption that CT slice thickness is 5 mm 
or less. If CT scans have slice thickness greater than 5 mm, the minimum 
size for a measurable lesion should be twice the slice thickness. 
• 
As with CT, if an MRI is performed, the technical specifications of the 
scanning sequences used should be optimized for the evaluation of the type 
and site of disease. The lesions should be measured on the same pulse 
sequence. Ideally, the same type of scanner should be used and the image 
acquisition protocol should be followed as closely as possible to prior scans. 
Body scans should be performed with breath-hold scanning techniques, if 
possible. 
• 
Physical Examination: For superficial non-nodal lesions, physical 
examination is acceptable, but imaging is preferable, if both can be done. In 
the case of skin lesions, documentation by color photography, including a 
ruler to estimate the size of the lesion, is recommended. 
11.33 Measurement at Follow-up Evaluation:  
• 
In the case of stable disease (SD), follow-up measurements must have met 
the SD criteria at least once after study entry at a minimum interval of 8 
weeks (see Section 11.433). 
• 
The cytological confirmation of the neoplastic origin of any effusion that 
appears or worsens during treatment when the measurable tumor has met 
criteria for response or stable disease is mandatory to differentiate between 
response or stable disease (an effusion may be a side effect of the treatment) 
and progressive disease. 
• 
Cytologic and histologic techniques can be used to differentiate between PR 
and CR in rare cases. 
11.4 
Measurement of Effect 
11.4.1 Target Lesions & Target Lymph Nodes 
• 
Measurable lesions (as defined in Section 11.21) up to a maximum of 
5 lesions representative of all involved organs, should be identified as 
“Target Lesions” and recorded and measured at baseline. These lesions can 
be non- nodal or nodal (as defined in 11.21), where no more than 2 lesions 
are from the same organ and no more than 2 malignant nodal lesions are 
selected. 
Note: If fewer than 5 target lesions and target lymph nodes are identified (as 
there often will be), there is no reason to perform additional studies beyond 
those specified in the protocol to discover new lesions. 
• 
Target lesions and target lymph nodes should be selected on the basis of their 
size, be representative of all involved sites of disease, but in addition should 
be those that lend themselves to reproducible repeated measurements. It may 
be the case that, on occasion, the largest lesion (or malignant lymph node) 
does not lend itself to reproducible measurements in which circumstance the 
next largest lesion (or malignant lymph node) which can be measured 
reproducibly should be selected. 
• 
Baseline Sum of Dimensions (BSD): A sum of the longest diameter for all 

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Amendment 7 
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target lesions plus the sum of the short axis of all the target lymph nodes will 
be calculated and reported as the baseline sum of dimensions (BSD). The 
BSD will be used as reference to further characterize any objective tumor 
response in the measurable dimension of the disease. 
• 
Post-Baseline Sum of the Dimensions (PBSD): A sum of the longest 
diameter for all target lesions plus the sum of the short axis of all the target 
lymph nodes will be calculated and reported as the post-baseline sum of 
dimensions (PBSD). If the radiologist is able to provide an actual measure 
for the target lesion (or target lymph node), that should be recorded, even if it 
is below 0.5 cm. If the target lesion (or target lymph node) is believed to be 
present and is faintly seen but too small to measure, a default value of 0.5 cm 
should be assigned. If it is the opinion of the radiologist that the target lesion 
or target lymph node has likely disappeared, the measurement should be 
recorded as 0 cm. 
• 
The minimum sum of the dimensions (MSD) is the minimum of the BSD and 
the PBSD.  
11.42 Non-Target Lesions & Non-Target Lymph Nodes 
Non-measurable sites of disease (Section 11.22) are classified as non- target 
lesions or non-target lymph nodes and should also be recorded at baseline. These 
lesions and lymph nodes should be followed in accord with 11.433. 
11.43 Response Criteria 
11.431 
All target lesions and target lymph nodes followed by 
CT/MRI/physical examination must be measured on re-evaluation at 
evaluation times specified in Section 11.1. Specifically, a change in 
objective status to either a PR or CR cannot be done without re- 
measuring target lesions and target lymph nodes. 
Note: Non-target lesions and non-target lymph nodes should be 
evaluated at each assessment, especially in the case of first response or 
confirmation of response. In selected circumstances, certain non-target 
organs may be evaluated less frequently. For example, bone scans may 
need to be repeated only when complete response is identified in target 
disease or when progression in bone is suspected. 
11.432 
Evaluation of Target Lesions 
• Complete Response (CR): All of the following must be true: 
a. Disappearance of all target lesions. 
b. Each target lymph node must have reduction in short axis to 
<1.0 cm. 
• 
Partial Response (PR): At least a 30% decrease in PBSD (sum of 
the longest diameter for all target lesions plus the sum of the short 
axis of all the target lymph nodes at current evaluation) taking as 
reference the BSD (see Section 11.41). 
• 
Progression (PD): At least one of the following must be true: 
a. At least one new malignant lesion, which also includes any 
lymph node that was normal at baseline (<1.0 cm short axis) 
and increased to ≥1.0 cm short axis during follow-up. 
b. At least a 20% increase in PBSD (sum of the longest diameter 
for all target lesions plus the sum of the short axis of all the 

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Amendment 7 
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target lymph nodes at current evaluation) taking as reference 
the MSD (Section 11.41). In addition, the PBSD must also 
demonstrate an absolute increase of at least 0.5 cm from the 
MSD. 
• 
Stable Disease (SD): Neither sufficient shrinkage to qualify for 
PR, nor sufficient increase to qualify for PD taking as reference the 
MSD. 
11.433 
Evaluation of Non-Target Lesions & Non-target Lymph Nodes 
• Complete Response (CR):  All of the following must be true: 
a. Disappearance of all non-target lesions. 
b. Each non-target lymph node must have a reduction in short 
axis to <1.0 cm. 
• Non-CR/Non-PD: Persistence of one or more non-target lesions or 
non-target lymph nodes. 
• Progression (PD): At least one of the following must be true: 
a. At least one new malignant lesion, which also includes any 
lymph node that was normal at baseline (<1.0 cm short axis) 
and increased to ≥1.0 cm short axis during follow-up. 
b. Unequivocal progression of existing non-target lesions and non-
target lymph nodes. (NOTE: Unequivocal progression should 
not normally trump target lesion and target lymph node status. 
It must be representative of overall disease status change.) 
11.44 Overall Objective Status 
The overall objective status for an evaluation is determined by combining the 
patient’s status on target lesions, target lymph nodes, non-target lesions, non-
target lymph nodes, and new disease as defined in the following tables: 
11.441 For Patients with Measurable Disease 
Target Lesions & Target 
Lymph Nodes 
Non-Target Lesions & Non-
Target Lymph Nodes 
New 
Sites of Disease 
Overall 
Objective 
Status 
CR 
CR 
No 
CR 
CR 
Non-CR/Non-PD 
No 
PR 
PR 
CR 
Non-CR/Non-PD 
No 
PR 
CR/PR 
Not All Evaluated* 
No 
PR** 
SD 
CR 
Non-CR/Non-PD Not All 
Evaluated* 
No 
SD 
Not all Evaluated 
CR 
Non-CR/Non-PD Not All 
Evaluated* 
No 
Not Evaluated 
(NE) 
PD 
Unequivocal PD 
CR 
Non-CR/Non-PD Not All 
Evaluated*
Yes or No 
PD 
CR/PR/SD/PD/Not all Evaluated 
Unequivocal PD 
Yes or No 
PD 

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Target Lesions & Target 
Lymph Nodes 
Non-Target Lesions & Non-
Target Lymph Nodes 
New 
Sites of Disease 
Overall 
Objective 
Status 
CR/PR/SD/PD/Not all Evaluated 
CR 
Non-CR/Non-PD Not All 
Evaluated* 
Yes 
PD 
*See Section 11.431 
** NOTE: This study uses the protocol RECIST v1.1 template dated 2/16/2011.  
11.442 For Patients with Non-Measurable Disease Only: 
Non-Target Lesions & 
Non-Target Lymph Nodes 
New Sites of Disease 
Overall Objective Status 
CR 
No 
CR 
Non-CR/Non-PD 
No 
Non-CR/Non-PD 
Not All Evaluated* 
No 
Not Evaluated (NE) 
Unequivocal PD 
Yes or No 
PD 
Any 
Yes 
PD 
*See Section 11.431 
11.45 Symptomatic Deterioration:  
Patients with global deterioration of health status requiring discontinuation of 
treatment without objective evidence of disease progression at that time, and not 
either related to study treatment or other medical conditions, should be reported 
as PD due to “symptomatic deterioration.” Every effort should be made to 
document the objective progression even after discontinuation of treatment due to 
symptomatic deterioration. A patient is classified as having PD due to 
“symptomatic deterioration” if any of the following occur that are not either 
related to study treatment or other medical conditions: 
• 
Weight loss >10% of body weight. 
• 
Worsening of tumor-related symptoms. 
• 
Decline in performance status of >1 level on ECOG scale. 
12.0 
Descriptive Factors 
12.1 
Dose level (as assigned by Registration Office): -2 vs -1 vs 1 vs 2 vs 3 vs 4 vs 5 
 
 

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13.0 
Treatment/Follow-up Decision at Evaluation of Patient 
13.1 
Initial Treatment 
Initial treatment is a single vaccine infusion followed by a 28-day follow-up period. 
Patients who develop PD, refuse, or initiate alternate treatment during this period should 
go to Event Monitoring. Patients who complete this period or who experience 
unacceptable adverse events during this period should go to Observation (see 13.2). 
13.2 
Observation 
After treatment completion patients will have one safety-observation visit at 29 days 
post-treatment. Observation will be every 3 months for 1 year, unless the patient 
progresses, refuses, or initiates alternate treatment at which time point the patient should 
go to Event Monitoring. 
13.3 
Replacement 
If a patient fails to complete the initial course of therapy (virus administration and 28 
days of follow-up) for reasons other than dose-limiting toxicity defined adverse events, 
the patient will be regarded as uninformative in regard to the primary study goal and an 
additional patient will be treated at the current dose level; however, all toxicity 
information will be utilized in the analysis.  
13.4 
Ineligible 
A patient is deemed ineligible if after registration, it is determined that at the time of 
registration, the patient did not satisfy each and every eligibility criteria for study entry. 
The patient may continue treatment at the discretion of the physician as long as there are 
no safety concerns, and the patient was properly registered. 
If treatment is stopped, the patient will go directly to the event-monitoring phase of the 
study (or off study, if applicable). 
• 
If the patient received treatment, all data up until the point of confirmation of 
ineligibility must be submitted. Event monitoring will be required per Section 4.0 of 
the protocol.  
• 
If the patient never received treatment, on-study material must be submitted. Event 
monitoring will be required per Section 4.0 of the protocol. 
13.5 
Major violation 
A patient is deemed a major violation, if protocol requirements regarding treatment in 
Cycle 1 of the therapy are severely violated that evaluability for primary end point is 
questionable. All data up until the point of confirmation of a major violation must be 
submitted. The patient will go directly to the event-monitoring phase of the study. Event 
monitoring will be required per Section 4.0 of the protocol. 
13.6 
Cancel 
A patient is deemed a cancel if he/she is removed from the study for any reason before 
any study treatment is given. On-study material and the End of Active Treatment/Cancel 
Notification Form must be submitted. No further data submission is necessary. 
 

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14.0 
Pharmacologic/Ancillary studies 
14.1 
Summary Table of Research Blood and Body Fluid Specimens to be Collected for this Protocol 
Correlative 
Study 
Mandatory 
or 
Optional 
Blood or 
Body 
Fluid 
being 
Collected 
Type of 
Collection 
Tube 
(color of 
tube top) 
Volume to 
collect per 
tube (# of 
tubes to be 
collected) 
After 
Reg 
prior 
to 
Txa 
Day 
1-2a 
Day 
3 
Day 
8 
Day 
10 
Day 
15 
Day 
29 
6 wks 
post 
VSV 
3 
mos 
post-
VSV 
6 
mos 
post-
VSV 
9 
mos 
post-
VSV 
12 
mos 
post-
VSV 
or 
EOTb 
Anti-VSV IgG 
and PRN 
Mandatory 
Blood 
Serum 
(red) 
6ml (1) 
X 
 
 
 
 
 
X 
 
 
 
 
 
Blood for viremia 
(PK) 
Mandatory 
Blood 
Paxgene 
2.5 ml (2) 
X 
X 
X 
X 
 
X 
O 
 
 
 
 
 
Plasma for 
cytokines, hIFNβ, 
virus level, and 
PBMC for 
sequencing, virus 
recovery & 
neoantigen 
Mandatory 
Blood 
Sodium 
heparin 
(green) 
10ml  
X (6) 
 
 
 
 
 
 
X (4) X (5) 
 
 
 
Citrate 
(blue) 
2.7ml (4) 
X 
X 
X 
X 
X 
O 
 
X 
X 
 
 
 
Plasma for ELISA 
assays, PBMC for 
cytokine and T-
cell immuno-
phenotyping 
assays (MB) 
Mandatory 
Blood 
Sodium 
heparin 
(green) 
10 ml  
X 
(18) 
 
 
 
 
X (2) 
X 
(18) X (2) 
X 
(18) X (2) X (2) X (18) 
Viral shedding - 
buccal swab 
Mandatory 
Cheek 
cells 
NA 
NA 
X 
X 
X 
X 
X 
X 
O 
 
 
 
 
 
Viral shedding - 
Mouth rinse 
Mandatory 
Mouth 
rinse 
NA 
50 ml (1) 
X 
X 
X 
X 
X 
X 
O 
 
 
 
 
 
Viral shedding - 
urine 
Mandatory Midstream 
urine 
Urine 
collection 
250ml (1) 
X 
X 
X 
X 
X 
X 
O 
 
 
 
 
 
X= required; O= optional additional testing if needed 
a. See table for Day 1-Day 2 collections in Section 14.11. Baseline samples can be collected at any time after registration and prior to 
infusion of virus. 
b. End of treatment/observation at any time (eg due to progression, new treatment, etc) 
 

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14.11 Day 1-Day 2 Sample Collections (before, during, and after infusion) 
Correlative 
Study 
Tube 
Prior to 
infusion 
At end 
of 
infusion 
30 
min 
after 
60 
min 
after 
2 hrs 
after 
4 hrs 
after 
Day 2:  
20-24 
hrs 
after 
Anti-VSV IgG 
and PRN 
Serum (red) 
(1 x 6ml) 
X 
 
 
 
 
 
 
Blood for 
viremia (PK) 
Paxgene  
(2 x 2.5ml) 
X 
X 
X 
X 
X 
X 
X 
PBMC for 
sequencing and 
T-cell assay 
NaHep 
(green) 
(5 x 10ml) 
X 
 
 
 
 
 
 
Citrate  
(blue 4 x 
2.7ml) 
X 
 
X 
X 
 
X 
X 
Human IFNβ 
and cytokine 
profile 
Sodium 
heparin 
(green)  
(1 x 10ml) 
X 
 
X 
X 
 
X 
X 
Buccal swab* 
NA 
X 
 
 
 
 
 
X 
Mouth rinse* 
50 ml (1) 
X 
 
 
 
 
 
X 
Midstream 
urine* 
250ml (1) 
collection 
container 
X 
 
 
 
 
 
X 
*Initial processing will be performed in CRTU.  
Med-Onc nurse will page laboratory 
 
for immediate pickup (See Section 14.33). 
14.2 
Collection and Processing  
14.21 Blood collection and processing 
Initial samples (Baseline, Day 1-2) will be collected in the hospital. When 
samples are available page 
 
for immediate pickup. 
Subsequent blood samples will be collected in CRTU. CRTU will page 
 
for immediate pickup. 
14.22 Virus shedding samples – buccal swab, mouth rinse, urine 
Initial samples will be collected in the hospital. Subsequent samples may be 
collected in CRTU in Charlton. 
When samples are available, page 
 
for immediate pickup. 

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14.3 
Shipping and Handling 
14.31 Kits will not be used for this study. 
14.32 Shipping Specimens – Not applicable 
14.33 Handling Specimens 
MedOnc staff will notify the following staff as soon as samples are available for 
pickup: page lab at 
for 
immediate pickup. 
Subsequent blood samples will be collected in clinical labs and transferred 
through BAP. BAP will page 
for immediate 
pickup. 
14.4 
Background and Methodology 
14.41 Assessment of viremia and viral shedding.  
Mononuclear cells will be isolated from blood, throat washings and urine from all 
study patients on Days 1-2, 3, 8 and 10. Additional samples on Days 15 and 29 
might be collected (depending on the viral RNA profile) and all will be tested for 
a) viral replication (quantitative RT-PCR to determine virus RNA copy number 
and co-culture on Vero cells for virus isolation). Testing will be performed in 
 
 
Viremia safety assay 
Whole blood collected into Paxgene tubes is stabilized and RNA will be 
extracted from whole blood. RNA will be subjected to quantitative RT-PCR to 
determine the replication of the virus over from, from point of infusion, its acute 
pharmacokinetic decline during first few hours, and its subsequent (if any) re-
appearance in the blood due to replication in the tumor cells. 
Viral Shedding Assays 
Buccal swabs, mouthwash or urine samples will be collected and the presence or 
absence of infectious virus or viral genomes will be analyzed using virus overlay 
rescue assays or qRT-PCR. These assays will help us evaluate if infectious virus 
is shed into body fluids after systemic administration into human subjects. 
14.42 Assessment of the peripheral immune response to viral administration. 
VSV specific immunity will be evaluated at baseline and on Day 29 by (a) 
measuring anti VSV specific antibodies (IgG) and plaque reduction 
neutralization assay (PRN). 
14.43 Evaluation of VSV-hIFNβ-NIS biodistribution using SPECT/CT or TFB-PET. 
See Section 7.19a. 
14.44 Immunophenotyping assays  
The systemic impact of treatment on immune cell subsets will be ascertained by 
immunophenotypic analysis of frozen PBMC for subsets of T helper cells, 
macrophages, and DC. Changes in Th1/Th2 ratio, DC1/DC2 ratio, and M1/M2 
ratio between the pre-treatment sample and each other sample will be used to 

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determine the systemic impact of treatment on immune cell subsets. Testing will 
be performed in 
 
14.45 Sequencing studies 
Sequencing will be performed to determine the whole exome and transciptome of 
the normal blood cells (PBMC) and tumor cells where possible. We will use 
bioinformatics to compare and contrast the sequences with the goal of identifying 
unique gene signatures that might indicate if a patient cancer might be especially 
susceptible or resistant to VSV virotherapy. It would also help us identify new 
antigens present in the tumor cells that might be useful as tumor specific markers. 
Testing will be performed in 
 
14.46 Immune profiling 
Extensive monitoring is performed in this trial to determine the baseline and post 
virus treatment immune profile of the patient. The blood collected will be used to 
identify the subset of immune cells present, or determine the immune reactivity 
of the T cells at baseline. Changes in the immune profile and phenotype will be 
analyzed using flow cytometry, immunostaining and various T cell specific 
assays to determine if there are tumor antigen reactive T cells. Testing will be 
performed in 
 
Pre- and post-treatment PBMCs will be viably frozen. The frequency of 
cytokine-secreting tumor antigen-specific T cells will be quantitated via ELISpot. 
In addition, pre- and post-treatment plasma samples will be assayed by ELISA 
for the emergence of antibodies against tumor antigens. Testing will be 
performed in 
 
14.47 Plasma cytokines and soluble PD-L1 
To assess for changes in systemic mediators of inflammation and 
immunosuppression, we will analyze concentrations of multiple cytokines in pre- 
and post-treatment plasma via electrochemiluminescence. We will also analyze 
concentrations of soluble PD-L1 via ELISA. We will compare pre- versus post-
treatment concentrations to determine whether PD-L1 or other mediators change 
in concentration upon treatment with VSV-hIFNβ-NIS. 
 

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15.0 
Drug Information 
15.1 
VSV-hIFNβ-NIS 
15.11 Background 
VSV-hIFNβ-NIS is a live, virus engineered to express both the human interferon 
β gene and the thyroidal sodium iodide symporter (NIS). The virus was 
constructed by inserting the gene for human IFNβ downstream of M gene and the 
NIS gene (cDNA) downstream of the gene for the G protein into a full-length 
infectious molecular clone of an Indiana strain VSV. This virus is not a vaccine. 
VSV-hIFNβ-NIS propagates on BHK cells with similar kinetics to the parental 
strain of virus and can be grown to high titers. It propagates selectively in human 
cancer cells since many of them cannot mount an effective antiviral response 
mediated via the IFN pathway. However, IFN production from infected cells will 
serve to protect noncancerous cells from the effects of the virus. As a result, the 
virus is directly cytopathic to tumor cells leading their rapid lysis with 
amplification of the virus. VSV-hIFNβ-NIS infected tumor cells also express 
NIS, a membrane ion channel that actively transports iodide into cells. 
Radioiodine uptake by cells expressing NIS provides the basis for in vivo imaging 
with 99mTc pertechnetate or radioiodine I-123 that can reveal the time dependent 
profile of VSV-hIFNβ-NIS gene expression and the location of VSV-hIFNβ-NIS 
infected cells during virus spread and elimination. 
15.12 Formulation 
The VSV-hIFNβ-NIS as manufactured (i.e., undiluted) is in a buffer consisting of 
5% sucrose, 50 mM Tris (pH 7.4), 2 mM MgCl2. After manufacture, the virus is 
stored frozen at ≤-65°C until prepared for use immediately prior to 
administration. 
15.13 Preparation and storage 
VSV-hIFNβ-NIS will be prepared at the Virus and Vector Production Laboratory 
(VVPL) of the Molecular Medicine Program at Mayo Clinic in Rochester MN. The 
virus clinical product is stored at ≤-65°C. The VSV-hIFNβ-NIS clinical product 
will be transferred in limited amounts to the Mayo Clinic Hospital - Methodist 
Campus Central Pharmacy (Eisenberg 1-406) and prepared in the Mayo Clinic 
Hospital -Methodist Campus Research Pharmacy () (Rochester, MN). 
The virus will be thawed and mixed with normal saline with 1% Human 
Serum Albumin (HSA) immediately prior to administration. The diluted 
virus is stable for 6 hours at room temperature. 
15.14 Administration 
See Section 7.0. 
15.15 Pharmacokinetic information 
{None available at this time} 
 

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15.16 Potential Drug Interactions 
{Unknown at this time} 
15.17 Known potential toxicities: 
VSV-hIFNβ-NIS has not been tested in the clinic and therefore, we do not know 
what the potential toxicities are. However, the virus was rescued from a derivative 
of the Indiana strain of VSV, so we anticipate that the worst toxicities will be 
similar to those experienced after infection of humans with VSV. However, the 
recombinant virus is attenuated compared to the parent Indiana strain due to 
addition of two transcriptional units encoding NIS and IFNβ, as well as IFNβ 
production by infected cells. In humans, the infection is mild or even 
asymptomatic. Typical symptoms include fever, headache, myalgia, malaise, 
nausea, vomiting and sometimes vesicles on the lips or mouth. The illness is 
usually short lived (less than 5 days) and requires only symptomatic treatment.  
As described in Sections 7.1 and 9.3, there is a risk of cytokine release syndrome 
(CRS). Symptoms suggestive of cytokine release syndrome include fever, chills, 
shaking hypotension, and/or symptoms of anaphylaxis such as dyspnea, itching, 
dizziness, or symptomatic hypotension. 
CRS can also lead to severe coagulopathy with prolonged prothrombin time or 
partial thromboplastin time and low fibrinogen levels. Inflammation from CRS 
and increased cytokine production can change hemodynamics, decreasing renal 
blood flow and glomerular filtration rate and potentially causing acute kidney 
injury and renal dysfunction. Some patients have also developed macrophage-
activation syndrome or hemophagocytic lymphohistiocytosis. 
There is one reported case of encephalitis that coincided with acute 
seroconversion to VSV. This occurred in a 3 year old child who was otherwise 
healthy. The child did survive the infection and was only treated with supportive 
care [6]. In vitro studies suggest that ribavirin can block VSV replication but we 
are not aware of any circumstances when this drug was used to treat acute VSV 
infection. We will be particularly vigilant for symptoms suggestive of 
meningoencephalitis that have been rarely observed. Such a patient will be 
treated aggressively with ribavirin and all the supportive care necessary as the 
situation might dictate.  
Because the effects on a fetus are unknown, recipients should not be pregnant at 
the time of receiving a virus or become pregnant for one year after receiving the 
virus and should not father a child for one year after receiving the virus.  
In addition, recipients should not donate sperm/ova, blood or blood products for 
one year after virus administration. 
15.18 Drug procurement 
Mayo Clinic Viral Vector Production Laboratory, located at Mayo Clinic in 
Rochester, MN, manufactures the investigational agent on site and transfers the 
product to the Research Pharmacy for final preparation. 
15.19 Nursing Guidelines 
VSV-hIFNβ-NIS testing in humans is limited and we do not know all possible 

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reactions. Acute reactions to injected virus and/or cytokine release syndrome 
(CRS) can be expected in immunocompetent patients. Peak time for such 
reactions is generally 4-6 hours after virus administration. Monitor for fever, 
nausea, and hypotension. Administer supportive care as described in Section 7.0 
and Section 9.0.Patients will be closely monitored during and after infusion to 
address any potential adverse events. 
15.2 
Ruxolitinib (INCB018424, INC424, Jakafi®) 
15.21 Background 
Ruxolitinib represents a novel, potent, and selective inhibitor of the JAKs with 
selectivity for JAK1 and JAK2. Ruxolitinib interferes with the signaling of a 
number of cytokines and growth factors that are important for hematopoiesis and 
immune function. 
15.22 Formulation 
Ruxolitinib tablets of 5 mg, 10 mg, 15 mg, 20 mg and 25 mg strengths have been 
developed as uncoated immediate release dosage forms for oral administration.  
The higher strength tablets are quantitatively proportional to the 5 mg tablets as 
all tablet strengths are compressed from a common blend. Excipients include: 
lactose monohydrate, cellulose microcrystalline, sodium starch glycolate (Type 
A), hydroxypropylcellulose, povidone, silica colloidal anhydrous, and 
magnesium stearate. 
15.23 Preparation and storage:  
Based on available stability data, the drug product should be stored in HDPE 
bottles with induction sealing and child-resistant closure between 15°C and 30°C, 
and should be protected from light. 
15.24 Administration 
Doses are taken approximately 12 hours apart without regard to food. 
15.25 Pharmacokinetic information: 
a) Absorption – near complete absorption, >95% in the GI tract. Mean peak 
plasma concentrations are achieved 1-2 hours post-dose. There was no clinically 
relevant change in the PK of ruxolitinib upon administration with a high-fat 
meal. Therefore, the drug may be administered either with or without food. 
b) Distribution – The apparent volume of distribution at steady-state (Vss/F) is 53-
65L in MF patients.  Plasma protein binding is approximately 97% in vitro, mostly 
to albumin. There is moderate distribution to organs and tissues with no long-term 
retention of drug-related material in preclinical species and limited drug penetration 
into the central nervous system (CNS) or across the blood-brain barrier. 
c) Metabolism – Extensive metabolism, with less than 1% of parent drug 
excreted in urine or feces. Metabolism is predominantly via the cytochrome P450 
isozyme CYP3A4 to yield oxygenated and subsequent conjugated metabolites. 
When administering ruxolitinib with strong CYP3A4 inhibitors, the total daily 
dose should be reduced by approximately 50%. 
d) Excretion – The mean terminal elimination half-life is ~3 h with no 
appreciable accumulation of either parent or metabolites with twice daily dosing.  
Metabolites are excreted in urine (74%) and feces (22%). 
15.26 Potential Drug Interactions 

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At clinically relevant concentrations, ruxolitinib and M18 (major metabolite in 
human circulation) are not anticipated to inhibit systemically CYP1A2, CYP2B6, 
CYP2C8, CYP2C9, CYP2C19, CYP2D6, CYP3A4, OAT1, OAT3, OCT1, 
OCT2, OATP1B1, OATP1B3, MDR1or MXR. However, an inhibition of 
intestinal CYP3A4, MDR1 and MXR by ruxolitinib cannot entirely be excluded. 
When administering ruxolitinib with strong CYP3A4 inhibitors, the total daily 
dose of ruxolitinib should be decreased by approximately 50% based on the 
platelet counts (or as specified in country-specific product labels). No dose 
adjustment is necessary when a mild or moderate CYP3A4 inhibitor is used as 
concomitant medication (although patients should be monitored closely for 
cytopenias when starting a mild or moderate CYP3A4 inhibitor). 
Upon initiation of a CYP3A4 inducer, no dose adjustment is recommended. 
Gradual dose increases of ruxolitinib may be considered if the effectiveness of 
therapy is diminished during chronic treatment with a CYP3A4 inducer. 
Patients with severe renal or hepatic impairment are not eligible for this trial.  
In patients with moderate renal impairment (Clcr = 30 -50 mL/min), the 
recommended starting dose based on platelet count should be reduced by 
approximately 50% to be administered twice a day. Ruxolitinib doses should be 
titrated based in individual safety and efficacy. (NOTE: We suggest starting with 
10mg twice per day for this protocol [Section 8.2].) 
In patients with mild or moderate hepatic impairment, the recommended starting 
dose based on platelet count should be reduced by approximately 50% with 
subsequent dose titration based on individual safety and efficacy. (NOTE: We 
suggest starting with 10mg twice per day for this protocol [Section 8.2].) 
Avoid grapefruit and grapefruit containing products while on study. 
15.27 Known potential adverse events 
Common known potential adverse events, >10%:  
Cardiovascular: Peripheral edema (22%) 
Central nervous system: Dizziness (15-18%), headache (10-15%), 
insomnia (12%) 
Dermatologic: Bruising (19-23%) 
Endocrine & metabolic: Cholesterol increased (17%) 
Gastrointestinal: Diarrhea (23%), constipation (13%), nausea (13%), 
vomiting (12%) 
Hematologic/Investigations: Anemia (96%), thrombocytopenia (70%), 
neutropenia (19%) 
Hepatic: ALT increased (25%), AST increased (17%) 
Respiratory: Dyspnea (16%), nasopharyngitis (16%) 
Less common known potential adverse events, 1-10%: 
Gastrointestinal: Flatulence (5%) 
Genitourinary: Urinary tract infection (9%) 
Miscellaneous: Herpes Zoster infection (2%) 
Rare, less than 1% (limited to important or life-threatening): 
Cardiac murmur, edema, peripheral neuropathy, withdrawal syndrome (acute 
relapse of myelofibrosis symptoms, splenomegaly, worsening cytopenias, 
hemodynamic compensation, and septic shock-like syndrome). 

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15.28 Drug procurement 
Ruxolitinib will be provided free of charge to study participants. 
15.29 Nursing Guidelines 
15.291 
Thrombocytopenia and anemia are common and appear to be dose 
related. Monitor CBC and instruct patients to report any unusual 
bruising or bleeding, or excessive fatigue to study team. 
15.292 
Neutropenia is less common, but is also dose related. Monitor CBC, 
and instruct patient to report any signs or symptoms of infection to 
study team. 
15.293 
Dizziness has been seen. Warn patient of this side effect. 
15.294 
Headache has also been seen. Treat symptomatically and monitor for 
effectiveness. 
15.295 
Monitor LFTs and discuss elevated levels with MD prior to 
administration. 
15.296 
Document all of patient’s medications, including OTC and herbals. 
When ruxolitinib is administered with strong CYP3A4 inhibitors the 
total daily dose of ruxolitinib should be reduced by approximately 
50%. Additionally hematologic monitoring should be increased when 
ruxolitinib and CYP3A4 inhibitors are administered together. 
15.297 
Assess patient’s baseline renal function and monitor per protocol. 
Patients with moderate (Clcr 30-50 mLmin) should have their starting 
dose reduced by 50% (based on platelet count) and administered twice 
daily (see section 15.26 for recommended dosing) 
 
 
 

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16.0 
Statistical Considerations and Methodology 
16.1 
Overview 
This is two parallel Phase I studies designed to determine in a sequential manner the 
MTD of VSV-hIFNβ-NIS (alone and in combination with ruxolitinib) when administered 
to immunocompetent patients with metastatic and/or incurable EC. The study follows the 
standard ‘cohort of three’ design used in Phase I clinical trials. Toxicity and 
pharmacokinetics (viral spread, expression and elimination) of VSV-hIFNβ-NIS in the 
treatment of EC will be evaluated at each of the dose levels. This section applies to both 
treatment groups, which will be analyzed separately.  
16.11 Maximum Tolerated Dose (MTD) 
The MTD will be defined as the highest safely-tolerated dose level where at most 
one patient out of six experiences DLT with the next higher dose level having at 
least 2 of 6 patients who have experienced DLT (see Section 7). 
16.12 MTD determination 
The MTD for all dose levels of VSV-hIFNβ-NIS (alone and in combination with 
ruxolitinib) will be determined as follows: if one patient experiences a DLT, up 
to three additional patients will be treated at the same dose level. If DLT is 
observed in only one of six patients treated at a given dose level, the next cohort 
of three patients will be treated at the next higher dose level. If two or more 
patients experience DLT at a particular dose level, then the dose escalation will 
cease and all subsequent patients will be treated at the lower dose level. 
16.13 Sample Size, Accrual and Study Duration 
As of February 2019, 9 patients have been enrolled in 3 cohorts of 3 patients 
(3 patients on Dose Level 1 and 6 patients on Dose Level 2) in Group A, prior to 
the modification of the DLT criteria in Amendment 4. At this time, Dose Level 2 
was restarted in Group A for enrollment of up to 6 additional patients to be 
evaluated under the new DLT criteria at Dose Level 2 prior to any dose 
escalation. Moving forward, with the addition of Group B, this study will involve 
additional patients. 
In Group A, we can expect a minimum of 12 patients (if the MTD is reached 
upon the enrollment of 3 additional patients at Dose Level 2) and a maximum of 
37 patients (3 patients at Dose Level 1, 6 patients Dose Level 2 prior to the new 
DLT criteria, 6 patients at each Dose Level post the new DLT criteria and 4 
additional patients to account for replacements for cancels, ineligibles and 
treatment violations). 
In Group B, we can expect a minimum of 9 patients (DLTs in at least 2 patients 
observed in first three patients at the starting dose level and 6 at dose level -1 if 
deemed MTD) and as many as 40 patients (6 patients at each dose level and 
4 additional patients to account for replacements for cancels, ineligibles, and 
treatment violations).  
Thus, the entire study will have as few as 21 patients and as many as 77 patients. 
Per year at MCR, ~15 new stage IV ECs undergo surgery and 20-30 cases of 
recurrent EC are evaluated. Assuming each patient cohort will require 
approximately 5 weeks of follow up for DLT after the last patient has been 
accrued (see Section 7.5), each cohort of 3 patients will require about 2 months 

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for accrual, treatment, and observation to determine incidence of DLTs and 
whether or not dose escalation can proceed. Therefore, the accrual period is 
expected to be at most 46 months from the date of Amendment 4, or 
approximately 64 months from study activation, taking into account the time needed 
to assess toxicity in each cohort. 
16.14 General Statistical Considerations 
The trial is designed to provide data about pharmacokinetics (biodistribution, 
targeting, viral gene expression and viral elimination), safety and biological 
activity of VSV-hIFNβ-NIS in patients with myeloma. Data related to toxicity 
and pharmacology will be presented using descriptive statistics due to the 
exploratory nature of the study. The data will be presented in table formats listing 
the mean, standard deviation and number of patients per group for continuous 
data, or listing count and percentages for categorical data as appropriate. All the 
relevant data will be used both in exploratory and hypothesis generating fashions 
to examine factors related to toxicity and pharmacology. 
16.2 
Analysis Plans  
16.21 Primary outcome analyses 
The number and severity of toxicity incidents will indicate the level of tolerance 
for VSV-hIFNβ-NIS (alone and in combination with ruxolitinib) as therapy for 
EC. For each of the stages, non-hematologic toxicities will be evaluated via the 
CTCAE v.4 standard adverse event grading. Hematologic toxicity measures 
such as anemia, neutropenia and thrombocytopenia will be assessed using 
continuous variables as the outcome measures (nadir and percent change from 
baseline values) as well as categorization via CTCAE v.4 standard adverse 
event grading. Frequency distributions and other descriptive measures will form 
the basis of the analysis of these variables. 
16.22 Secondary endpoints 
The following secondary endpoints will be evaluated for each stage 
independently. In addition, differences in dose levels with and without 
cyclophosphamide may also be explored where appropriate. 
16.221 
Clinical Response 
The number of clinical responses may provide useful preliminary data 
on the efficacy of this treatment regimen in this patient population. A 
clinical response in this setting will be defined as noted in Section 11.0. 
The number of responses (CR, PR, or SD) will be summarized by 
simple descriptive summary statistics across all patients in each group 
as well as by dose level and primary type of cancer (EC). 
16.222 
Correlative studies 
Data will be collected for a number of laboratory correlative variables 
as discussed before (e.g. viral replication and shedding). Descriptive 
statistics and scatterplots will form the basis of presentation of these 
variables. Correlations between the laboratory values and other 
outcome measures will be carried out by standard parametric and non-
parametric tests (e.g. Pearson’s and Spearman’s rho). Data obtained 
from NIS imaging of VSV- hIFNβ-NIS will be used to determine the 

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biodistribution and kinetics of virus spread and NIS gene expression in 
vivo and correlate it with tumor distribution. 
16.223 
Tolerability 
Tolerability of this regimen will be explored in an ancillary manner 
through time-related variables including time until any treatment related 
toxicity, time until treatment related Grade 3+ toxicity and time until 
hematologic nadirs (WBC, ANC, platelets). Simple summary statistics 
will be supplemented with Kaplan-Meier survival estimates and related 
confidence intervals. The effect of dose and ancillary dichotomized 
covariates such as age will be explored using logrank testing involving 
one covariate at a time. Again the small sample size restricts the 
generalizability of such testing, but the results will provide preliminary 
indications for subsequent research in Phase II clinical trials. 
16.23 Monitoring 
The principal investigator(s) and the study statistician will review the study 
continually during enrollment to identify accrual, adverse event, and any 
endpoint problems that might be developing.  
The Mayo Clinic Cancer Center (MCCC) Data Safety Monitoring Board 
(DSMB) is responsible for reviewing accrual and safety data for this trial at least 
twice a year, based on reports provided by the MCCC Statistical Office. 
16.24 Adverse Event Stopping Rules 
NOTE: If the stopping rules below are met, subject enrollment will be stopped 
until subject cases are submitted to the FDA for review and discussion prior to 
re-starting subject enrollment. 
16.241 For Group B only (patients receiving ruxolitinib in combination with 
VSV-hIFNβ-NIS) 
- 
Within a group (across dose levels), more than 2 patients with 
Grade 4 neutropenia lasting more than 7 days at least possibly related 
to study treatment during first 28 days 
- 
Within a group (across dose levels), more than 2 patients with 
Grade 4 thrombocytopenia lasting more than 7 days at least possibly 
related to study treatment during the first 28 days 
16.242 For Group A and B 
- 
Any Grade 5 event at least possibly related to study treatment during 
first 28 days 
16.3 
Results Reporting on ClinicalTrials.gov 
At study activation, this study will have been registered within the “ClinicalTrials.gov” 
website. The Primary and Secondary Endpoints along with other required information for 
this study will be reported on 
For purposes of timing of the 
Results Reporting, the estimated completion date for the Primary Endpoint of this study 
is 45 months after the study opens to accrual. The definition of “Primary Endpoint 
Completion Date” (PECD) for this study is at the time the last patient registered has been 
followed for at least 6 months. 

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16.4 
Subset Analyses for Women and Minorities 
16.41 Inclusion of women and minorities 
By the nature of the cancers being studied, only women will be included in this 
trial. This study will be available to all eligible patients, regardless of race or 
ethnic origin. There is no information currently available regarding differential 
effects of this regimen in subsets defined by race and there is no reason to expect 
such differences to exist. Therefore, although the planned analyses will, as 
always, look for differences in treatment effect based on racial groupings, the 
sample size is not increased to provide additional power for subset analyses. 
16.42 Enrollment of women 
Women will comprise 100% of this patient population secondary to the nature of 
the cancers being studied as only occurring in women.  
16.43 Expected minority enrollment 
In prior Mayo Clinic studies in this disease, approximate 3% of all patients were 
classified as ethnic minorities. Therefore, it is likely that only 1 or 2 patients 
classified as ethnic minorities will be enrolled in this trial. 
16.44 Ethnicity/Race/Gender Table 
Expected sizes of ethnicity and race by gender subsets are shown in the following 
table: 
Accrual Targets 
Ethnic Category 
Sex/Gender 
Females 
Males 
Total 
Hispanic or Latino 
3 
0 
3 
Not Hispanic or Latino 
74 
0 
74 
Ethnic Category: Total of all subjects 
77 
0 
77 
Racial Category 
American Indian or Alaskan Native 
0 
0 
0 
Asian 
0 
0 
0 
Black or African American 
3 
0 
3 
Native Hawaiian or other Pacific Islander 
0 
0 
0 
White 
74 
0 
74 
Racial Category: Total of all subjects 
77 
0 
77 
 
Ethnic 
Categories: 
Hispanic or Latino – a person of Cuban, Mexican, Puerto Rican, South or 
Central American, or other Spanish culture or origin, regardless of race. The 
term “Spanish origin” can also be used in addition to “Hispanic or Latino.” 
Not Hispanic or Latino 
Racial 
Categories: 
American Indian or Alaskan Native – a person having origins in any of the 
original peoples of North, Central, or South America, and who maintains tribal 
affiliations or community attachment. 
Asian – a person having origins in any of the original peoples of the Far East, 
Southeast Asia, or the Indian subcontinent including, for example, Cambodia, 

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China, India, Japan, Korea, Malaysia, Pakistan, the Philippine Islands, 
Thailand, and Vietnam. (Note: Individuals from the Philippine Islands have 
been recorded as Pacific Islanders in previous data collection strategies.)  
Black or African American – a person having origins in any of the black 
racial groups of Africa.  
Native Hawaiian or other Pacific Islander – a person having origins in any of 
the original peoples of Hawaii, Guam, Samoa, or other Pacific Islands. 
White – a person having origins in any of the original peoples of Europe, the 
Middle East, or North Africa. 
 
 

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17.0 
Pathology Considerations 
Biopsies of accessible lesions will be collected at specified intervals for research purposes in connection with the study. Attempts 
will be made to avoid biopsying RECIST target lesions when feasible.  
17.1  
Summary Table of Research Tissue Specimens to be Collected for this Protocol 
Correlative Study 
(Section for more 
information) 
Mandatory 
or Optional 
Type of 
Tissue to 
Collect 
Block, Slides, 
Core, etc. (# of 
each to submit) 
Pre-
Treatment2 
After any 
NIS imaging 
scan 
Day 
29 
3 mos post 
infusion 
Temperature 
Conditions for 
Storage 
/Shipping 
Tumor biopsy 
Mandatory1 
Frozen 
2 cores 
X 
 
 
 
frozen 
Tumor biopsy 
Mandatory1 
FFPE 
4 cores 
X 
 
X 
X 
ambient 
NIS-lit tissue biopsy3 
Mandatory1 
Frozen and 
RNALater 
and FFPE  
3 cores  
(1 of each 
type) 
 
X 
 
 
 
1. If feasible and accessible. If patient refuses, there is no deviation. 
2. Pre-treatment specimen can be from endometrial cancer tissue repository or Tissue Registry 
3. Biopsy if NISimaging shows positive areas. Biopsy may occur within 3 days after any positive scan. 
17.2 
Correlative Tissue Collection  
17.21 Tissue Kits will not be provided for this protocol. Page  
PRIOR to biopsy to collect 
samples from procedure room. 
17.22 Frozen Tissues 
17.221 
Place biopsy into clean sterile tube and freeze immediately on dry ice or -20°C freezer.  
17.222 
Page 
to collect for processing for infectious virus recovery and exome 
sequencing 
17.22 RNA Later Tissues 
17.231 
Place biopsy into tube containing RNAlater solution and leave at room temperature.  
17.232 
Page 
to collect for processing for RNA sequencing and RT-PCR for viral 
genomes 
 

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17.23 FFPE Tissues 
17.231 
Place biopsy cores into 10% formalin and paraffin embed one core into 
one block. 
17.232 
Return paraffin embedded blocks to 
 
for immunohistochemical staining of immune cell markers and T 
cell infiltrates. 
17.233 
NIS positive biopsy block will be given to 
for staining of viral antigens and NIS protein 
17.3 
Background and Methodology 
Perform RNAseq and exome sequencing on tumor and normal tissues. 
Hypothesis: The 84 gene signature set can serve as a theranostic test in future VSV trials 
to predict susceptibility of the tumor to VSV replication and associated viremia and/or 
response. 
a) Perform RNAseq and analysis of tumor pre-treatment, and after VSV (where 
possible), 
b) Perform exome sequencing on tumor and PBMC pre-treatment to identify somatic 
mutation rate and potential neoantigens, 
c) Assess pharmacokinetics of VSV viremia (virus in blood)  
d) Assess viral shedding and persistence in urine and respiratory secretions,  
e) Assess tumor selectivity of VSV-hIFNβ-NIS by assaying for VSV in tumor and 
normal tissues, and assess serum IFNβ levels post VSV administration. 
Sequencing studies 
Sequencing will be performed to determine the whole exome and transciptome of the 
normal blood cells (PBMC) and tumor cells where possible. We will use bioinformatics 
to compare and contrast the sequences with the goal of identifying unique gene signatures 
that might indicate if a patient cancer might be especially susceptible or resistant to VSV 
virotherapy. It would also help us identify new antigens present in the tumor cells that 
might be useful as tumor specific markers. 
Assess the impact of IV VSV-hIFNβ-NIS on adaptive immune responses against 
VSV and EC antigens and on immunosuppressive mediators.  
Hypotheses: 1) Treatment with VSV-hIFNβ-NIS will lead to an increased host immune 
response against EC antigens. 2) Understanding the nature of the immunosuppressive 
mediators that impact the immunogenicity of VSV-hIFNβ-NIS will guide the design of 
rational therapeutic combinations in EC with VSV IV.  
a) Quantitate humoral responses to VSV-hIFNβ-NIS and candidate EC antigens, 
b) Quantitate T cell responses to VSV-hIFNβ-NIS and EC antigens, and 
c) Assess immunosuppressive mediators impacting the immunogenicity of VSV-
hIFNβ-NIS. 
Quantitate humoral responses to VSV-hIFNβ-NIS and candidate EC antigens. 
Determine EC tissue expression of FRα, p53, survivin, HER-2, and TERT. For each 
patient treated with VSV-hIFNβ-NIS, we will perform immunohistochemistry analysis of 
pre-treatment EC biopsy samples for FRα, p53, survivin, HER-2, and TERT using 
techniques that have been established by the Mayo Clinic Department of Laboratory 
Medicine and Pathology. Tumors will be scored as positive or negative for expression of 
each antigen. 

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Assess immunosuppressive mediators impacting the immunogenicity of VSV-
hIFNβ-NIS. 
Determine changes in tumor-infiltrating leukocyte subsets after VSV-HIFNβ-NIS 
treatment. We will perform immunofluorescence staining for CD8, CD4, CD25, and 
FoxP3 on EC biopsies from before and after VSV-hIFNβ-NIS treatment. We will 
quantitate the number of CD8+, CD4+, and CD4+25+FoxP3+ (Treg) cells and compare 
pre-treatment versus post-treatment TIL frequencies and correlate the number of CD8 
TILs with tumor response as measured by RECIST 1.1 criteria. 
Determine changes in regional immunosuppressive pathways. We will perform 
immunohistochemistry analysis for PD-L1 and IDO on EC biopsies from before and after 
VSV-hIFNβ-NIS treatment. We will compare pre-treatment versus post-treatment 
staining for IDO and PD-L1 to determine whether a change is induced by VSV-hIFNβ-
NIS treatment. 
 

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18.0 
Records and Data Collection Procedures 
18.1 
Submission Timetable 
Data submission instructions for this study can be found in the Data Submission 
Schedule. 
18.2 
Event monitoring 
See Section 4.0 and Data Submission Schedule for the event monitoring schedule. 
18.3 
CRF completion 
This study will use Medidata Rave® for remote data capture (rdc) of all study data. 
18.4 
Site responsibilities 
Each site will be responsible for insuring that all materials contain the patient’s initials, 
MCCC registration number, and MCCC protocol number. Patient’s name must be 
removed. 
18.5 
Supporting documentation 
This study requires that AE assessment used to determine eligibility be uploaded into 
Supporting Documents. This study requires supporting documentation for evidence of 
response to study therapy and progression after study therapy.  
18.6 
Incomplete materials 
Any materials deemed incomplete by the MCCC Operations Office will be considered 
“not received” and will not be edited or otherwise processed until the missing 
information is received. A list of the missing documents will be made available to the 
appropriate co-sponsor/participant. 
18.7 
Overdue lists 
A list of overdue materials and forms for study patients will be generated monthly. The 
listings will be sorted by location and will include the patient study registration number. 
The appropriate co-sponsor/participant will be responsible to obtain the overdue material. 
18.8 
Corrections forms 
If a correction is necessary the QAS will query the site. The query will be sent to the 
appropriate site to make the correction and return the query and documentation of 
correction back to the QAS.  
 
 

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19.0 
Budget Considerations 
19.1 
Costs charged to patient 
Routine clinical care 
19.2 
Hospital inpatient expenses 
Patients enrolled in the study will not be billed for room and board or nursing charges 
while in the Medical Oncology Unit at Mayo Clinic Hospital – Methodist Campus. 
However, participants may be billed for ancillary expenses such as any oral medications 
prescribed at the time of discharge. 
19.3 
Tests that will be research funded 
The following items will be covered by research funds: 
• 
HIV, hepatitis and tuberculosis testing at baseline 
• 
Optional NIS imaging by Planar and SPECT/CT scans or TFB-PET (if done) 
• 
Optional tumor biopsies  
• 
Tests for viral status and shedding post-infusion including tests of immune 
response to the virus 
• 
Study treatment with VSV-NIS-hIFNβ and ruxolitinib 
 
 

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20.0 
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Appendix I 
ECOG Performance Status 
 
ECOG PERFORMANCE STATUS* 
Grade ECOG 
0 
Fully active, able to carry on all pre-disease performance without restriction 
1 
Restricted in physically strenuous activity but ambulatory and able to carry out 
work of a light or sedentary nature, e.g., light house work, office work 
2 
Ambulatory and capable of all selfcare but unable to carry out any work 
activities. Up and about more than 50% of waking hours 
3 
Capable of only limited selfcare, confined to bed or chair more than 50% of 
waking hours. 
4 
Completely disabled. Cannot carry on any selfcare. Totally confined to bed or 
chair. 
5 
Dead 
*As published in Am. J. Clin. Oncol.: 
Oken, M.M., Creech, R.H., Tormey, D.C., Horton, J., Davis, T.E., McFadden, E.T., Carbone, P.P.: 
Toxicity And Response Criteria Of The Eastern Cooperative Oncology Group. Am J Clin Oncol 5:649-
655, 1982. 
The ECOG Performance Status is in the public domain therefore available for public use. To duplicate the 
scale, please cite the reference above and credit the Eastern Cooperative Oncology Group, 
 
 
 
 

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Appendix II 
New York Heart Association Classification of Congestive Heart Failure 
 
NYHA 
Class 
Symptoms 
I 
Cardiac disease, but no symptoms and no limitation in ordinary physical activity, e.g. no 
shortness of breath when walking, climbing stairs etc. 
II 
Mild symptoms (mild shortness of breath and/or angina) and slight limitation during ordinary 
activity. 
III 
Marked limitation in activity due to symptoms, even during less-than-ordinary activity, e.g. 
walking short distances (20–100 m). Comfortable only at rest. 
IV 
Severe limitations. Experiences symptoms even while at rest. Mostly bedbound patients. 
 
Adapted from Dolgin M, Association NYH, Fox AC, Gorlin R, Levin RI, New York Heart Association. 
Criteria Committee. Nomenclature and criteria for diagnosis of diseases of the heart and great vessels. 
9th ed. Boston, MA: Lippincott Williams and Wilkins; March 1, 1994. 
Original source: Criteria Committee, New York Heart Association , Inc.  Diseases of the Heart and Blood 
Vessels. Nomenclature and Criteria for diagnosis, 6th edition  Boston, Little, Brown and Co. 1964, p 114. 
 

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Appendix III Child-Pugh Score 
The score employs five clinical measures of liver disease. Each measure is scored 1-3, with 3 indicating 
most severe derangement.[1] 
Measure 
1 point 
2 points 
3 points 
Total bilirubin, μmol/L 
(mg/dL) 
<34 (<2) 
34-50 (2-3) 
>50 (>3) 
Serum albumin, g/dL 
>3.5 
2.8-3.5 
<2.8 
Prothrombin time, 
prolongation (s) 
<4.0 
4.0-6.0 
>6.0 
Ascites 
None 
Mild (or suppressed with 
medication) 
Moderate to Severe (or 
refractory) 
Hepatic encephalopathy 
None 
Grade I-II 
Grade III-IV 
Chronic liver disease is classified into Child-Pugh class A to C, employing the added score from above.[1] 
Points Class One year survival Two year survival 
5-6 
A 
100% 
85% 
7-9 
B 
81% 
57% 
10-15 
C 
45% 
35% 
References 
1. 
Cholongitas, E; Papatheodoridis, GV; Vangeli, M; Terreni, N; Patch, D; Burroughs, AK (Dec 
2005). "Systematic review: The model for end-stage liver disease--should it replace Child-Pugh's classification 
for assessing prognosis in cirrhosis?". Alimentary pharmacology & therapeutics. 22 (11-12): 1079–
89. doi:10.1111/j.1365-2036.2005.02691.x. PMID 16305721. 
2. 
Child CG, Turcotte JG (1964). "Surgery and portal hypertension". In Child CG. The liver and portal 
hypertension. Philadelphia: Saunders. pp. 50–64. 
3. 
Pugh RN, Murray-Lyon IM, Dawson JL, Pietroni MC, Williams R (1973). "Transection of the oesophagus for 
bleeding oesophageal varices". The British journal of surgery.60 (8): 646–
9. doi:10.1002/bjs.1800600817. PMID 4541913. 
 

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Appendix IV Study Drug Diary 
Document removed - The study drug diary is provided as a standalone document as required by the Mayo 
Clinic Foundation Institutional Review Board (IRB). This version is removed to avoid the appearance of 
conflicting versions in the case of audit.