A Pilot Study to Test Whether Systemic Interferon Gamma Increases Tumor Class I MHC Expression in Patients With Synovial Sarcoma and Myxoid/Round Cell Liposarcoma
Phase: EARLY_PHASE1
Conditions: Myxoid Liposarcoma, Round Cell Liposarcoma, Synovial Sarcoma
Sponsor: Fred Hutchinson Cancer Center

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D o c u m e nt R el e a s e d D at e 

Protocol 2705.00: Interferon Gamma for SS and MRCL 
1. INTRODUCTION 
This protocol tests whether systemic interferon gamma (IFNy) will increase class I 
MHC expression in patients with advanced myxoid/round cell liposarcoma (MRCL) and 
synovial sarcoma (SS). These patients historically have a very poor prognosis with a 
median survival of less than one year [1]. Our group has been developing T cell therapy for 
these patients, however a major barrier is the low MHC expression of these tumors. 
Identification of a systemic agent that up regulates MHC in these tumors is critical for 
successful development of T cell therapy. IFNy increases MHC expression in vitro in these 
tumors and has been observed to increase MHC in vivo in melanoma tumors. 
Patient taking part in this study will have had tumor biopsy samples taken before 
entering the study and tissue will be removed after IFNy administration to determine 
whether there has been a change in class I MHC expression. In order to obtain post 
treatment tumor samples, this trial has been designed to enroll patients with metastatic 
disease undergoing resection, however, certain patients undergoing resection of primary 
tumors and patients who have disease easily accessible for a core needle biopsy at bedside 
may also be eligible. 
IFNy is an FDA approved drug with an established safety profile that has been well 
studied in cancer patients with multiple studies dating back to the early 1980's. However, it 
is not currently considered standard of care for any cancer type. 
1.1 
Introduction to the Prevalence, Biology and Natural History of Synovial 
Sarcoma and MRCL 
Soft-tissue sarcomas cause approximately 3900 deaths annually including a 
disproportionate number of deaths in children, adolescents and young adults. The median 
survival for patients with advanced soft tissue sarcoma is 8 to 12 months and only 20 to 
25% of patients are alive at 2 years [2]. 
This protocol is focused on 2 soft tissue sarcoma subtypes: synovial sarcoma (SS) 
and myxoid/ round cell liposarcoma (MRCL). SS has an incidence close to 800 cases 
annually and represents about 8% of soft tissue sarcomas. It has peak incidence in the 3rd 
decade of life 13,4]. SS is by definition associated with a characteristic t(X;18) translocation 
resulting in one of the SYT-SSX fusion proteins [5]. Liposarcomas account for 
approximately 10-20% of soft tissue sarcomas and can be classified into three subtypes each 
with their own distinct clinical behaviors: pleomorphic, well/de-differentiated and myxoid/ 
round cell liposarcoma (MRCL). MRCL accounts for 40-50% of liposarcomas and is almost 
always associated with a chromosomal translocation, most commonly t(12;16)(q13;p11) 
though a number of less common translocations have also been described [6]. The resultant 
fusion proteins have an activity that is not well understood [7]. 
While chemotherapy sensitive sarcoma subtypes including MRCL and SS may 
respond to first line chemotherapy in a minority of cases (typically Adriamycin and 
Ifosfamide), in almost all patients, the disease will ultimately progress. Pazopanib is a small 
molecule tyrosine kinase inhibitor with potent anti-VEGF activity that has been approved 
for patients with SS and other metastatic soft tissue sarcomas generally (with the exception 
of liposarcomas such as MRCL). In a phase III clinical trial pazopanib improved 
progression free survival benefit (PFS) of sarcoma patients by approximately 3 months but 

Before 
10 weeks 
after 
• 
Figure 1: Response to NY-ES0-1 specific T cells. 
Patient responded in several lung tumor including 
large RUL tumor (A and B) however a soft tissue 
tumor progressed in spite of therapy (C and D). NY-
ES0-1 was retained (E and F). Class I MHC 
expression was absent before and after treatment (G 
and H). 
Protocol 2705.00: Interferon Gamma for SS and MRCL 
did not demonstrate a statistically significant overall survival benefit (OS). There are no 
other FDA approved agents for patients with metastatic soft tissue sarcomas [8]. 
Trabectedin is a drug that has shown promise for liposarcomas and a national phase III trial 
is currently evaluating its benefit (NCT00210665). However, responses are limited in 
duration and mortality remains high, suggesting the need for novel approaches [9]. 
1.2 
Expression of NY-ESO-1 in Synovial Sarcoma and MRCL 
NY-ES0-1, a member of the family of Cancer Testis Antigens, was first discovered 
through serological analysis in esophageal cancer patients and was subsequently found to 
induce a strong cytotoxic T-cell response [10, 11]. As their name implies, CT Antigens, are 
expressed on a protein level in various malignant tumors and germ cells of the testis but not 
other adult tissues. 
Based on its immunogenicity, NY-ESO-1 is considered to be among the most 
attractive targets for immunotherapy. It has been targeted in a number of clinical studies 
including several vaccine trials that have induced serologic, CD4+ and CD8+ T cell 
responses. Delayed type hypersensitivity responses following NY-ESO-1 vaccination have 
been associated with long-term survival [12, 131. Objective clinical responses have been 
observed in melanoma patients following 
vaccination against NY-ESO-1, including one 
complete response [14]. T cell therapies 
targeting NY-ESO-1 have induced both partial 
and complete responses [15, 16]. 
A vast majority of synovial sarcomas 
have been described as expressing NY-ES0-1. 
In an analysis from Jungbluth et al., 20 of 25 
(80%) synovial sarcoma tumors were found to 
express NY-ESO-1 by IHC. Four of the five 
tumors not expressing NY-ESO-1 had the 
biphasic synovial sarcoma subtype suggesting 
an even higher proportion of NY-ES0-1 
expression among the monophasic subtype. 
Eight of the 20 NY-ESO-1 expressing tumors 
had homogenous expression [17]. More 
recently, we have found that MRCL also has a 
strong prevalence of NY-ESO-1 expression 
[18]. We tested 25 MRCL tumors and found 
that all expressed NY-ESO-1 (100%) and that 
expression was homogenous over 70% of cases. 
[16]. 
13 

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Figure 2: Class I MHC in an A0201+ cell line established in 
our lab increases from baseline (red) following treatment 
with IFNy 100 U/mL (blue). Expression is further increased 
at the 1000U/mL concentration (green) although this 
concentration in probably supra physiologic. NY-ES0-1 
specific T cell killing is also increased following treatment 
at the 100U/mL concentration (B). Gp100 (not expressed by 
this cell line) specific T cells are used as a control for non-
antigen specific tumor lysis. 
Protocol 2705.00: Interferon Gamma for SS and MRCL 
13 
Experience Treating Synovial Sarcoma and MRCL with NY-ESO-1 Specific 
Effectors 
Robbins et al. reported on the clinical outcomes of 6 synovial sarcoma patients [16]. 
Four of the patients had partial responses however none of the responses lasted longer than 
1 year. One of the responding patients was treated with a second infusion after progression 
following a partial response, when the two responses were combined together the total 
length of response was 18 months. An intensive regimen was used including pre infusion 
high dose cyclophosphamide (CY) and fludarabine along with high dose IL-2 post infusion. 
Our group has used a strategy of isolating NY-ES0-1 specific T cells from the 
peripheral blood of SS and MRCL patients. Our first patient treated with antigen specific T 
cell therapy had significant anti-tumor responses (Figure 1 A and B) however the response 
was short lived and not all tumors responded despite persistence of NY-ES0-1 specific T 
cells (Figure 1 C and D). Sampling of a progressing tumor demonstrated homogenous 
strong NY-ESO-1 expression (E and F) but absent class I MHC (G and H) suggesting that 
this is a potential mechanism of immune evasion for these tumors. 
1.4 
MHC expression in sarcoma tumors 
We have found that MRCL tumors have decreased expression of both Class I and 
Class II MHC molecules. Over 50% of tested patients (13/25) had MHC class I expression 
that was either undetectable or only focally positive (see Appendix A). Another 36% has 
MHC class I expression that was only "1+" positive meaning that it was in less than 25% of 
cells examined. None of the tumors examined had over 50% of cells express MHC class I. 
MHC II data is similarly decreased. We have also looked for MHC expression in SS 
tumors. On a tissue microarray stained for class I MHC including 12 SS, seven tumors 
tested had absent MHC and the other 5 had very weak or focal class I MHC expression. 
Each of the 4 SS patients who have enrolled on our T cell therapy trial have had their 
tumors tested for class I MHC and all have been negative. 
13 
Increasing MHC expression in SS 
and MRCL cell lines using low dose 
IFNy 
We have now tested 2 MRCL cell 
lines and 3 SS cell lines with IFNy in vitro 
and seen up-regulation of class I MHC in 
each cell line tested. In all but one of the 
cell lines, marked increases in class II 
MHC were also observed. One of the SS 
cell lines, established in our lab from a 
potential T cell therapy patient, expressed 
the HLA type A0201. We were able to 
increase NY-ES0-1 specific killing by 
treating this cell line with IFNy at a 
physiologic concentration 100 U/mL 
(Figure 2) readily achievable in vivo [19, 
201. 
- - 4 - - 

Protocol 2705.00: Interferon Gamma for SS and MRCL 
1.6 
Increased MHC expression in melanoma tumors using low dose IFNy 
A number of agents have been observed to increase MHC expression in vitro in 
tumor cell lines. For example, both IFN alpha and gamma have both been shown to 
increase MHC class I and class II molecules in vitro in multiple cancer types in numerous 
studies [21]. We have also seen increased MHC expression in SS and MRCL cell lines 
following IFN alpha. However, we are not aware of any clinical data demonstrating that 
systemic administration of IFN alpha increases either class I and II MHC molecules for 
patients receiving typical doses. 
By contrast, IFNy has been shown to up regulate MHC molecules at low doses 
(0.1mg/m2  once weekly) [22] in as little as 1 week in melanoma tumors. MHC evaluation 
was based on the presence of positive cells in fine needle aspirates of accessible lesions. 
Two patients in this study had negative MHC class I expression in their tumor and both 
developed MHC I expression following therapy. For one patient, class I expression was 
induced after only 1 week of therapy however for the other patient it took 3 months. In 6 of 
14 patients starting without class II expression, MHC class II was induced. To be sure, 
melanoma and soft tissue sarcoma have different biologies and interact with the immune 
system in markedly different ways thus the need to test for this effect in SS and MRCL 
patients specifically. 
1.7 
Toxicities of IFNy treatment 
IFNy has been used extensively in clinical trials for patients with melanoma both in 
the metastatic as well as adjuvant settings. Currently it is FDA approved for patients with 
chronic granulomatous disease as well as a treatment for severe osteoporosis. Response 
rates for patients with metastatic melanoma treated with IFN gamma have ranged from 5-
15% with including a handful of complete responders [22, 23]. IFNy has been found to be 
safe and relatively well tolerated even at doses significantly higher than we are proposing to 
use in this study. 
A large phase I study treated 98 patients with metastatic melanoma at doses up to 
0.9 mg/m2  administered IV (rather than subcutaneously) three times per week for at least 8 
weeks. This study had no fatalities but did see several cases of grade 4 toxicities including 
anemia, myalgias, renal (proteinuria) and heaptic toxicity at higher doses. There were also 3 
cardiac events that occured in patients on that trial. 
A phase III randomized trial was run by SWOG in the early 1990's designed to test 
the benefit of IFNy in the adjuvant setting [24]. This trial included 133 patients who 
completed 1 year of daily IFN gamma and were evaluated for toxicity. There were no fatal 
complication or grade 4 toxicities, however, 24 patients had grade 3 toxicities including 4 
patients with grade 3 myelosuppression, 3 patients with grade 3 neurologic toxicities 
(confusion, insomnia, personality change) as well as other toxicities including depression, 
migrane, transaminitis, pruritus, and flu like symptoms. The most common toxicities were 
fever/chills and headache. It is unclear at what point of the year of therapy patients 
developed the grade 3 toxicities. Dosing in this trial gave each patient 0.2 mg of IFNy 
subcutaneously daily, similar to the 0.1mg/m2  considered to be low dose. Although the 

Protocol 2705.00: Interferon Gamma for SS and MRCL 
authors of the study concluded that single agent IFN gamma had no clinical benefit for 
melanoma patients in the adjuvant setting, it was safe and generally well tolerated. 
The low doses of IFNy proposed in this study (0.1mg/m2  weekly) were examined in 
19 patients who received up to six months of therapy. No patients on this study had grade 3 
or grade 4 toxicity. The most common toxicity were flu-like symptoms (grade 1). Grade 2 
toxicities included fever (2 patients) as well as chills, myalgias, flu-like symptoms and 
leukopenia (1 patient each). IFNy has also been used in prior trials safely in the 
perioperative period both right before surgery [25] and after surgery [26]. 
1.8 
Fatality in a patient receiving NY-ESO-1 specific T cells with IFNy 
The first two SS patient treated at the FHCRC with NY-ES0-1 specific T cells 
received low dose weekly IFNy. The first patients results are shown in Figure 1. The second 
synovial sarcoma patient treated with autologous NY-ES0-1 specific T cells in 
combination with IFNy died of sudden cardiac death. This patient also received 
Cyclophosphamidc and IL-2 concurrently with the IFNyas conditioning. This patient had 
uncontrolled diabetes with a hemoglobin Al C >10%, mild renal dysfunction at baseline 
and had recently been started on metformin. 
Autopsy ruled out a myocardial infarction or a pulmonary embolism. Analysis of the 
heart showed cardiomegaly (500 g), as well as selective myocyte necrosis and myocarditis 
suggesting both a metabolic derangement and myocardial inflammation, which may or may 
not have been related to each other. The cause of the potential metabolic abnormality is 
unclear as the only evaluation of his electrolytes was after the patient had been undergoing 
resuscitation for well over an hour. However, this may be a result of the patient's treatment 
with metformin and/or his underlying renal insufficiency. 
The myocardial infiltrate was overwhelmingly CD68+ and CD4+ cells. Since, the T 
cell product was >98% CD8+ T cells and NY-ES0-1 was not expressed by the myocardial 
tissue at autopsy the death was thought most likely related to the patients co-morbidities and 
conditioning, and unlikely to be directly related to the autologous T cell product itself. 
Interferon gamma and IL-2 have been removed in order to focus more closely on the safety 
of the T cell product. 3 patients have now safely been treated with the NY-ESO-1 specific T 
cells, though it is too soon to determine their clinical out comes. We believe that NY-ESO-1 
specific T cells can likely be safely combined with IFNy but this requires further study. For 
this reason, patients who have received NY-ESO-1 specific T cells are excluded from this 
study. If we find in this study that IFNy does indeed increase MHC expression in SS and 
MRCL tumors, a follow up study will be specifically designed to address the safety 
considerations of combining T cell therapy with IFNy. 
1.9 
Surgery in patients with metastatic sarcoma 
In order to test whether IFNy increases class I MHC expression, we have chosen to 
include patients in a number of different clinical scenarios. Primary tumors where there is a 
recent biopsy specimen available can participate (so long as the patient has not received 
systemic therapy or radiation since the biopsy was performed). Patients with metastatic 
disease who have a painful untreated primary tumor they would like remove may 
participate. Patients with lesions accessible to bed side core needle biopsy will be allowed 

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Protocol 2705.00: Interferon Gamma for SS and MRCL 
to participate. However, our expectation is that one of the most common situations for 
patients considering this trial are patients with oligometastatic lung metastasis considering 
resection. 
When metastatic disease is operable, removal is frequently considered the optimal 
choice. While a randomized control trial comparing a surgical approach to a non-surgical 
approach would be impossible, retrospective analysis and historical controls seem to 
suggest a benefit. For example, in a large analysis performed by the MSKCC [27], overall 
survival of sarcoma patients undergoing resection for metastatic disease was significantly 
increased relative to a similar historical group of patients (15 vs. 11 months). Isolated cases 
of patients with long-term disease free survival following metastetectomy have been 
reported [28]. In patients where complete resection can be achieved, survival at 3 years has 
been reported above 30%, particularly when the burden of disease is low [29]. Patients with 
a significant disease free interval or who relapse with only a small volume of disease after 
an initial metastatectomy may benefit from a second surgery [30]. Radiotherapy is another 
option for patients with a few isolated sarcoma metastasis [31]. 
Despite the potential benefits of surgery for some patients with metastatic sarcoma, 
there are patients who will not benefit. Some patients will rapidly progress despite surgery 
and undergo a major operation without any clinical benefit from it. Standard practice is to 
wait and observe the patient either with or without systemic therapy in order to assess the 
pace of disease before bringing the patient to surgery in order to avoid performing an 
unnecessary procedure on a patient with rapidly progressing 
disease [32-34]. It is during this waiting period that we 
expect to treat the majority of patients on this study. 
1.10 Results of the study to date 
To date we treated six patients; all evaluable biopsy 
specimens showed dramatically increased MHC expression; 
by flow cytometry, HLA-ABC expression rose from a mean 
of 3.3% to 17.0% (Fig. 2A and B). Markedly increased 
CD3+ cell infiltration was also observed (Fig. 2B). In order to 
interrogate the specificity of these T cells, TIL were 
expanded from each 1-2 mm core samples and using an 
ELIspot assay consisting of overlapping 15 amino acid 
peptides (Fig. 2C) from the most immunogenic regions of 
cancer testis antigens (including NY-ESO-1), we found that 
post-treatment T cells had improved recognition of these 
antigens. Thus, a single inflammatory cytokine can 
dramatically alter each component of the TME state. We are 
very pleased that based on our results a Cancer 
lmmunotherapy Trials Network Study is being planned 
combining IFNy with pembrolizumab. However, even though 
we believe that the study has met its primary end point and 
demonstrated increased MHC expression following IFNy 
treatment in these sarcoma tumors, we are seeking to treat 
up to four additional patients in order to have better data 
regarding our secondary objectives, specifically to study the 
PD-L1+ macrophages, which we have observed increased in 
patients treated on the study to date. 

Protocol 2705.00: Interferon Gamma for SS and MRCL 
2. OBJECTIVES 
2.1 
Primary Objectives 
• 
To determine whether systemic administration of IFNy will increase class I 
MHC expression in SS and MRCL tumors. 
2.2 
Secondary Objective 
• 
To determine whether systemic administration of IFNy will increase class II 
MHC expression in SS and MRCL tumors. 
• 
To examine changes in the immune response to MRCL and SS by 
examining changes in the immune infiltrates, antibody response and antigen specific 
T cell response before and after IFNy treatment. 
3. 
STUDY DESIGN 
This protocol will test whether systemic IFNy will increase class I MHC expression 
in patients with advanced myxoid/round cell liposarcoma (MRCL) and synovial sarcoma 
(SS). 
Patients with available prior tissue may use this as their pre-treatment sample. 
Patients may also have a research biopsy prior to receiving IFNy. For their post IFNy 
sample, patients may either have surgery or biopsy as part of their standard care or have a 
research biopsy. 
Ten patients will receive 4 weeks of low dose weekly IFNy at a dose of 100 
mcg/m2. If there are any issues regarding data from any of these patients, additional patients 
may be enrolled as replacements as the discretion of the PI. Patients will be given a patient 
log marked with the scheduled dates for IFNy administration. They will be instructed to 
record the day and time of IFNy administration in their logs .Deviations of up to 48 hours in 
administration of IFNy will be permitted while on the weekly schedule and up to 24 hours 
while on the three times weekly schedule. The final tumor sample should be taken 24-48 
hours after the last dose of IFNy and not more than 72 hours. 
While not ideal, the length of time that a patient is on IFNy with either regimen 
could be extended to up to 8 weeks to accommodate the scheduling changes sometimes 
necessary in the course of patient care. The patient may also stop IFNy and restart to 
accommodate a scheduling delay. Any such change in IFNy must be approved by the PI. 
If there is a patient who would be a good study candidate except that surgery is 
scheduled (or is being scheduled) too soon for them to receive 4 weekly doses of IFNy, but 
they could receive at least 2 doses of IFNy, they may participate and receive only 2 doses. 
While response to IFNy is not an endpoint of the study and no imaging is planned to 
evaluate response, should a clinical response to IFNy be noted through the normal course of 
clinical care, it is conceivable that a situation would develop where it was determined that 
continuation of IFNy will clinically benefit the patient (e.g. if the patient's tumor appears to 
be responding). In this case, IFNy may be continued at the discretion of the PI and the 
treating physician. 
For research biopsies, ideally 4-6 cores will be collected. Unless otherwise specified 
by the PI, one will be placed in formalin for IHC. The other samples should be placed in 
- - 8 - - 

Protocol 2705.00: Interferon Gamma for SS and MRCL 
RPM for analysis by the Pollack Lab. Biopsies may be done in the clinic at the SCCA if 
deemed safe and appropriate by the treating clinician. Alternatively, biopsies may be done 
under ultrasound guided at the University of Washington. 
Data from the study including patient outcomes and biomarker related information learned 
from analysis of patient samples may be shared with Horizon Pharma who will now be 
providing drug for the study. 
4. 
PATIENT ELIGIBILITY 
4.1 
Inclusion Criteria 
4.1.1 
A diagnosis of synovial sarcoma and myxoid/ round cell liposarcoma 
4.1.2 Male or female subject, 18 or older 
4.1.3 A superficial tumor easily and safely accessible for a research biopsy 
or are being considered for resection or biopsy of their tumor as part 
of standard of care and have recent pathology. 
4.1.4 Zubrod performance status of '0-2' or Karnofsky score >60%. 
(Appendix A) 
4.1.5 No treatment with systemic anti-cancer treatment (chemotherapy or 
biologics) within 2 weeks of starting interferon gamma. 
4.1.6 Patients with a history of coronary artery disease must have had a 
normal stress test within 180 days of starting IFNy. 
4.1.7. Must have been off metformin for at least 2 weeks prior to starting 
IFNy. 
4.1.8 No use of full dose, therapeutic anti-coagulation. However, low dose 
warfarin for catheter prophylaxis or acetylsalicylic acid are 
acceptable. 
4.1.9 No thrombotic event within 6 months (deep vein thrombosis, 
pulmonary embolism) of starting IFNy. 

Protocol 2705.00: Interferon Gamma for SS and MRCL 
4.2 
Exclusion Criteria 
4.2.1 
Active infection requiring oral or intravenous antibiotics 
4.2.2 Pregnant women, nursing mothers, men or women of reproductive 
ability who are unwilling to use effective contraception or 
abstinence. Women of childbearing potential must have a negative 
pregnancy test within two weeks prior to entry. 
4.2.3 Serum creatinine > 1.5 mg/dL or Glomerular Filtration Rate <50. 
4.2.4 Significant hepatic dysfunction (SGOT > 150 IU or > 3x upper limit 
of normal; bilirubin > 1.6 mg/dL; prothrombin time > 1.5 x control). 
4.2.5 Known CNS metastasis. Once CNS metastasis have been treated 
these patients may participate if they are otherwise good trial 
candidates. 
4.2.6 Current treatment with steroids (must be discontinued 1 week before 
starting IFNy). 
4.2.7 Hemoglobin A1C >8.5%. 
4.2.8 Uncontrolled hypertension, BP >150/100mmHg. Patients with 
elevated BP may enroll once BP is corrected. 
4.2.9 NY-ESO-1 specific T cell therapy within 1 year of starting on the 
trial. 
4.2.10 New (<6 months) cardiac arrhythmia (EKG should be performed 
within 2 weeks of starting IFNy. 
4.2.11 History of clinically significant congestive heart failure. 
5. STUDY AGENT 
Low Dose Weekly IFNy 
Interferon gamma lb (Actimmune) is FDA approved for the treatment of Chronic 
Granulomatous Disease and Osteopetrosis. Although it has been extensively studied for the 
treatment of cancer, it is not FDA approved for the treatment of cancer and is not 
considered a standard treatment for patients with sarcoma. 
IFNy will be given by sub-cutaneous administration at a dose of 0.1mg/ne on one of the 
two schedules described in the study design section prior to surgery. Patients or their 
caregiver will be instructed on subcutaneous self-administration. 
6. INVESTIGATIONAL PLAN 
All patients will have available tissue prior to study entry. The first two patients will 
receive low dose IFNy (0.1mg/m2) by sub-cutaneous administration at a dose of 0.1mg/m2 
for four weeks prior to surgery. Injection should be given every 7 days though an allowance 
of 2 days before or after will be acceptable to accommodate specific circumstance. Any 
deviation from the recommended schedule of IFNy dosing must be discussed with the PI. 
Patients or their caregiver will be instructed on subcutaneous self-administration. The last 
--10-- 

Protocol 2705.00: Interferon Gamma for SS and MRCL 
dose of IFNy will be given 24-48 hours prior to surgery (absolutely not more than 72 
hours). 
10 patients will be treated on this regimen. 
Paraffin embedded tissue will be tested for class I MHC, class II MHC and NY-ESO-1. 
Immune infiltrates in tumors will be studied using IHC staining for CD3, CD4, CD8, 
CD57, CD68 and FoxP3. Antibody responses will be examined by Sacha Gnjatic (Tisch 
Cancer Center). In patients with the HLA types A0201 and A2402 antigen specific T cell 
responses to NY-ES0-1 will be examined using MHC tetramers in the peripheral blood. In 
patients with the HLA type A0201, PRAME, MAGE family and CAMEL specific T cell 
responses will also be examined using MHC tetramers. In patients will large tumors that are 
resected, immune infiltrates will be examined directly by flow cytometry for markers of T 
cell memory (CD45RO, CD27, CD28, CCR7, CD62L, CD127) and when possible look for 
tumor infiltrating antigen specific T cells using MHC tetramers. When enough tumor is 
available RNA will also be extracted for possible future analysis. 
Toxicity will be recorded and graded according to the NCI CTCAE v 4.0 at each visit. 
7. MANAGEMENT OF TOXICITIES AND COMPLICATIONS 
7.1 
Criteria for Discontinuation of Therapy 
Toxicity grading will be evaluated according to guidelines in NCI Common 
Toxicity Criteria version 4.0 [36]. Therapy will be discontinued in the following 
circumstances: 
1. life threatening toxicity that is potentially a result of IFNy 
2. toxicity resulting in hospitalization 
3. Grade 3 toxicity 
4. Unexpected Grade 2 or greater toxicity 
5. Expected Grade 2 toxicity lasting for over 48 hours at Grade 2 or requiring 
hospitalization with the exception of grade 2 cytopenias which may last >48 hours. 
Expected Toxicities: 
i. 
Flu-like symptoms (headache, muscle ache, joint ache, chills) 
Redness, pain and swelling at injection site 
fatigue 
iv. 
rash 
v. 
nausea/emesis 
vi. 
abdominal pain 
vii. 
diarrhea 
viii. 
fever 
ix. 
cytopenia (anemia, lymphopenia, leukopenia may last >48 hours) 

Protocol 2705.00: Interferon Gamma for SS and MRCL 
If expected toxicities progress to grade 3 (including cytopenias), the patient will be 
discontinued from study. 
Patients discontinued from the study will not be considered in any subsequent data analysis 
other than toxicity data. 
7.2 
Concomitant Therapy 
All medical problems that should occur on the trial should be treated according to the 
standard of care. Patients are not allowed to have anti-cancer therapy (chemotherapy, 
radiation, biologics) while on study. If a patient developed any medical issues that require 
steroids, this will be reported in all analysis of data. 
73 
Premature Discontinuation 
Subjects who do not complete the study medication will be considered to have prematurely 
discontinued the study. The reasons for premature discontinuation (for example, voluntary 
withdrawal, toxicity, death) must be reported on the case report form. A subject may re-
enter the study after premature discontinuation only by approval of the Principal 
Investigator. If possible, final study evaluations should be completed at the time of 
discontinuation. Potential reasons for premature discontinuation include: 
73.1 The development of a life-threatening infection. 
73.2 Judgment of the principal investigator that the patient is too ill to 
continue. 
733 Patient noncompliance with study therapy and/or clinic 
appointments. 
73.4 Pregnancy. 
73.5 Voluntary withdrawal; a patient may remove himself/herself from 
the study at any time without prejudice. 
73.6 Significant and rapid progression of sarcoma requiring alternative 
medical or surgical intervention including, but not limited to, the 
development of CNS metastasis. 
73.7 Toxicity as per section 7.1. 
73.8 
Termination of the study by the principal investigator, Institutional 
Review Office or the Food and Drug Administration. 
8. SCHEDULE OF EVALUATIONS 
--12-- 

Protocol 2705.00: Interferon Gamma for SS and MRCL 
8.1 
General Toxicity Assessment 
Patients will have vital signs taken, a physical exam, a comprehensive chemistry panel and 
a complete blood count and differential at each planned visit. Visits will occur prior to 
starting IFNy, every other week while on IFNy, and two weeks after surgery. All adverse 
events > grade 2 will be recorded and graded according to the NCI CTCAE v 4.0. 
8.2 
Assessment of Tumor 
Paraffin embedded tumor will be tested for MHC and other studies using IHC (see Section 
6 Investigational Plan). If sufficient tumor is available some will be taken for RNA and T 
cell infiltrates will be expanded from the tumor for analysis. If still more tumor is available 
tumor cell lines may be attempted to be established and infiltrates may be analyzed by flow 
cytometry directly at the direction of the PI. 
8.3 
Schedule of Evaluations 
History and Physical Exam: Before starting 1FNy, 24-48 hours after first IFNy dose, every 2 
weeks while on IFNy, within 48 hours prior to surgery (if not already part of the every other 
weekly evaluations), 2 weeks following surgery. 
CBC with differential and platelet count, CMP, Hepatic panel, and research labs — With 
each history and physical exam 
Tumor tissue — to be obtained at time of surgery or biopsy 
Research labs will typically include 40mL in green top (heparinized) tubes and 10 mL in 
red top (non-heparinized) tubes and will typically be drawn only at each history and 
physical exam. However, up to 60 mL in green top tubes and up to 20 mL in red top tubes 
may be drawn as often as every week if there is a scientific rationale to sample more blood 
for an individual patient at the discretion of the PI. 

Protocol 2705.00: Interferon Gamma for SS and MRCL 
9. 
REPORTING ADVERSE EVENTS 
All unexpected and serious adverse events that may be due to study treatment or 
intervention must be reported to the FHCRC Institutional Review Office as soon as 
possible but within at least 10 calendar days of the investigator learning of the event. 
9.1 
Definition of an Adverse Event 
Any untoward medical occurrence in a patient or clinical investigation subject 
administered a pharmaceutical product, medical treatment or procedure and which does 
not necessarily have to have a causal relationship with this treatment. 
1. Life-threatening Adverse Event 
Any adverse event that places the patient, in view of the investigator, at 
immediate risk of death from the reaction. 
2. 
Unexpected Adverse Event 
An unexpected adverse event is an adverse event that is not described in the 
study protocol or informed consent. 
3. Serious Adverse Event (SAE) 
An SAE is any adverse event occurring that results in any of the following 
outcomes: 
• death, 
• a life-threatening adverse event (real risk of dying), 
• inpatient hospitalization or prolongation of existing hospitalization, 
• a persistent or significant disability/incapacity, 
• a congenital anomaly, 
• requires intervention to prevent permanent impairment of damage. 
9.2 
Attribution 
• Related — includes adverse events that are definitely, probably, or possibly related to 
the medical treatment or procedure. 
• Not Related — includes adverse events are doubtfully related or clearly not related to 
the medical treatment or procedure. 
9.3 
Procedure for Reporting Serious Adverse Events 
The FHCRC Serious Adverse Event (SAE) Report Form will be completed for all serious 
adverse events (unexpected which may be related to study treatment) that meet the 
expedited reporting requirements. The SAE form will be faxed to the IRO at (206) 667-
6831. All available information should be submitted but it is acceptable to fax an 
incomplete report form at the initial report. A completed report should be faxed as soon as 
possible but must be received within 10 calendar days. 
--14-- 

Protocol 2705.00: Interferon Gamma for SS and MRCL 
Unexpected serious adverse events that do not meet the requirement for expedited reporting 
must be reported to the IRB as part of the annual renewal of the protocol. 
Serious Adverse Events with also be reported to Horizon Pharma, who will now be 
supplying Interferon Gamma for the study. 
9.4 
Evaluation and Reporting of Adverse Events: 
Patients are monitored for the development of end organ damage by assessing adverse 
events with scrum chemistries, liver function studies, complete blood counts, and physical 
exams performed accordingly as explained in Section 8.3. 
We will report any adverse events related to IFNy that are unexpected (see expected 
toxicities in 7.1), serious (grade 2 or higher) or result in the discontinuation of therapy. See 
Section 7.1 regarding criterion for discontinuation of therapy. IFNy is an approved drug that 
has been used in hundreds of cancer patients and has an established record of expected 
toxicities. All adverse events for all systems arc graded on a scale of 1-5 and attribution is 
assigned, using the NCI Common Terminology Criteria for Adverse Events (version 4). 
10. 
STATISTICAL CONSIDERATIONS 
To date, 6 patients were treated with IFNy at a dose of 100mcg/m2, five of whom 
had adequate biopsy collection for MHC evaluation. All had a marked increase in MHC. 
However, we have also observed increased tumor infiltration with PD-L1+, CD11B+ cells. 
We are now increasing the planned enrollment of the study to 10 patients. For these 
additional four patients. it would be highly relevant to observe marked increase in these 
macrophages (effect size >2.5). Four patients gives us over 90% power to detect such a 
large increase with a two-tailed alpha of 0.05. These additional patients will also allow for 
more detailed visual illustration of CD3+ infiltration and MHC expression in anticipation 
of an up-coming follow-ip study. 
11. 
ADMINISTRATIVE CONSIDERATIONS 
11.1 
Institutional Review Board 
In accordance with federal regulations (21 CFR 312.66), an Institutional Review Board 
(IRB) that complies with regulations in 21 CFR 56 must review and approve this protocol 
and the informed consent form prior to initiation of the study. 
11.2 
Consent 
The Principal Investigator or his associate must explain verbally and in writing the nature, 
duration, and purpose of the study and possible consequences of treatment. Patients must 
also be informed that they may withdraw from the study at any time and for any reason 
without jeopardizing their future treatment. In accordance with federal regulations (21 
--15-- 

Protocol 2705.00: Interferon Gamma for SS and MRCL 
CFR 312), all patients must sign the IRB-approved consent form in the presence of a 
witness. 
11.3 Termination of Study 
The Principal Investigators reserve the right to terminate this study at any time. The FDA 
may also terminate the study. 

Protocol 2705.00: Interferon Gamma for SS and MRCL 
APPENDIX A 
ECOG / Zubrod Performance Status 
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 self-care but unable to carry out any 
work activities. Up and about more than 50% of waking hours 
3 
Capable of only limited self-care, confined to bed or chair more than 
50% of waking hours 
4 
Completely disabled. Cannot carry on any self-care. Totally confined 
to bed or chair 
5 
Dead 
Karnofsky Performance Status 
100 
Normal no complaints; no evidence of disease. 
90 
Able to carry on normal activity; minor signs or symptoms of disease. 
80 
Normal activity with effort; some signs or symptoms of disease. 
70 
Cares for self; unable to carry on normal activity or to do active work. 
60 
Requires occasional assistance, but is able to care for most of his 
personal needs 
50 
Requires considerable assistance and frequent medical care. 
40 
Disabled; requires special care and assistance. 
30 
Severely disabled; hospital admission is indicated although death not 
imminent. 
20 
Very sick; hospital admission necessary; active supportive treatment 
necessary. 
10 
Moribund; fatal processes progressing rapidly. 
0 
Dead 
--17-- 

Protocol 2705.00: Interferon Gamma for SS and MRCL 
APPENDIX B 
DATA AND SAFETY MONITORING PLAN 
The study will comply with the requirements of the FHCRC Data and Safety Monitoring 
Plan (DSMP). 
REPORTING ADVERSE EVENTS  
Definitions: 
Adverse Event: Any harm or untoward medical occurrence in a research participant 
administered a medical product, medical treatment or procedure even if it does not 
necessarily have a causal relationship with the product, treatment, or procedure. An 
adverse event can be any unfavorable and unintended sign (including an abnormal 
laboratory finding, for example), symptom, or disease temporally associated with the 
use of a medical product, medical treatment or procedure whether or not it is 
considered to be related. 
• An adverse event is "unexpected" when its specificity and severity are not 
accurately reflected in the informed consent document. 
• An adverse event is "related to the research procedures" if in the opinion of 
the principal investigator, it was more likely than not caused by the research 
procedures or if it is more likely that not that the event affects the rights and 
welfare of current participants). 
Life-threatening Adverse Event — Any adverse event that places the patient or 
subject, in view of the investigator, at immediate risk of death from the reaction. 
Serious Adverse Event (SAE) — Any adverse event occurring that results in any of 
the following outcomes: 
• death 
• a life-threatening adverse event (real risk of dying) 
• inpatient hospitalization or prolongation of existing hospitalization 
• a persistent or significant disability/incapacity 
• a congenital anomaly 
• requires intervention to prevent permanent impairment of damage 
Expedited Reporting Requirements to the IRB 
All unexpected and serious adverse events, which may be due to study treatment or 
intervention, must be reported to the FHCRC Institutional Review Office per their 
current reporting requirements. 

Protocol 2705.00: Interferon Gamma for SS and MRCL 
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