Modified White Blood Cells That Secrete IL-2 and Express a Protein That Targets the ESO-1tumor Protein for Metastatic Cancer

Background:

- In prior trials we have shown that adoptive transfer of lymphocytes, transduced with a T
cell receptor (TCR) that recognizes the NY-ESO-1 antigen, can mediate regression of
metastatic cancer, though responses are often transient, and complete responses are
rare.

- Interleukin-12 (IL-12) is an important immunostimulatory cytokine. We have constructed a
retroviral vector that contains an inducible single chain IL-12 driven by an NFAT
responsive promoter which can be used to mediate transfer of this gene into anti-tumor
lymphocytes. This construct enables the secretion of IL-12 following stimulation of the
TCR.

- Transduction of the IL-12 gene into mouse anti-tumor lymphocytes results in a profound
increase in the ability of these lymphocytes to mediate tumor regression following
administration to tumor bearing mice. These cells have a profound advantage in inducing
anti-tumor responses because very few cells are needed and there is no requirement for
the concomitant administration of interleukin-2 (IL-2) as is the case for conventional
cell transfer immunotherapies.

- Based on these studies we have now used a retrovirus that encodes an inducible human
single chain IL-12 driven by an NFAT responsive promoter and a retrovirus that encodes
an anti-NY-ESO-1 TCR to cotransduce autologous lymphocytes for the treatment of patients
with metastatic cancer that expresses the NY-ESO-1 antigen.

Objectives:

Primary objectives:

- To evaluate the safety of the administration of IL-12 and anti-NY-ESO-1 engineered
lymphocytes in patients receiving a non-myeloablative conditioning regimen.

- To determine if the administration of the transduced lymphocytes to patients following a
nonmyeloablative but lymphoid depleting preparative regimen will result in clinical
tumor regression in patients with metastatic cancer.

Secondary objective:

- To determine the in vivo survival of cotransduced gene-engineered cells.

Eligibility:

Patients who are 18 years of age or older must have

- Metastatic cancer whose tumors express the ESO antigen;

- ECOG performance status 0 or 1;

Design:

- Autlogous lymphocytes will be cotransduced with retroviral vectors, encoding IL-12 and
anti- NY-ESO-1 TCR.

- Patients will receive a nonmyeloablative but lymphocyte depleting preparative regimen
consisting of cyclophosphamide and fludarabine followed by intravenous infusion of
IL-12/anti-ESO TCR gene-transduced lymphocytes.

- Patients will undergo complete evaluation of tumor with physical examination, CT of the
chest, abdomen and pelvis and clinical laboratory evaluation four to six weeks after
treatment. If the patient has SD or tumor shrinkage, repeat complete evaluations will be
performed every 1-3 months. After the first year, patients continuing to respond will
continue to be followed with this evaluation every 3-4 months until off study criteria
are met.

- The study will be conducted using a Phase I/II optimal design. The protocol will proceed
in a phase 1 dose escalation design. Initially, the protocol will enroll 1 patient in
each of the first 3 dose cohorts unless that patient experiences a dose limiting
toxicity (DLT). Following cohort 3, all subsequent cohorts will proceed in a phase 1
dose escalation design, with 5 cohorts of n=3. Should a single patient experience a DLT
due to the cell transfer at a particular dose level, additional patients would be
treated at that dose to confirm that no greater than 1/6 patients have a DLT prior to
proceeding to the next higher level. If a level with 2 or more DLTs in 3-6 patients has
been identified, three additional patients will be accrued at the next-lowest dose, for
a total of 6, in order to further characterize the safety of the maximum tolerated dose
(MTD) prior to starting the phase II portion. If a DLT occurs in the first cohort, that
cohort will be expanded to 6 patients. If 2 DLTs are encountered in this cohort, the
study will be terminated.

- Once the MTD has been determined, the study then will proceed to the phase II portion
using a phase II optimal design where initially 21 evaluable patients will be enrolled
in each of 2 cohorts. If 0 or 1 of the 21 patients experiences a clinical response, then
no further patients will be enrolled but if 2 or more of the first 21 evaluable patients
enrolled have a clinical response, then accrual will continue until a total of 41
evaluable patients have been enrolled.

- The objective will be to determine if the combination of lymphocyte depleting
chemotherapy, and IL-12/ESO TCR gene engineered lymphocytes is associated with a
complete response rate that can rule out 5% (p0=0.05) in favor of a modest 20% PR + CR
rate (p1=0.20).

- INCLUSION CRITERIA:

1. Metastatic cancer that expresses ESO as assessed by one of the following methods:
RT-PCR on tumor tissue, or by immunohistochemistry of resected tissue, or serum
antibody reactive with ESO. Metastatic cancer diagnosis will be confirmed by the
Laboratory of Pathology at the NCI.

2. Patients with melanoma or renal cell cancer must have previously received high
dose IL-2 and have been either non-responders (progressive disease) or have
recurred. Patients with other histologies, must have previously received systemic
standard care (or effective salvage chemotherapy regimens) for metastatic
disease, if known to be effective for that disease, and have been either
non-responders (progressive disease) or have recurred.

3. Greater than or equal to 18 years of age.

4. Willing to sign a durable power of attorney

5. Able to understand and sign the Informed Consent Document

6. Clinical performance status of ECOG 0 or 1.

7. Life expectancy of greater than three months.

8. Patients of both genders must be willing to practice birth control from the time
of enrollment on this study and for up to four months after receiving the
preparative regimen.

9. Patients must be HLA-A*0201 positive

10. Serology:

1. Seronegative for HIV antibody. (The experimental treatment being evaluated
in this protocol depends on an intact immune system. Patients who are HIV
seropositive can have decreased immune-competence and thus be less
responsive to the experimental treatment and more susceptible to its
toxicities.)

2. Seronegative for hepatitis B antigen, and seronegative for hepatitis C
antibody. If hepatitis C antibody test is positive, then patient must be
tested for the presence of antigen by RT-PCR and be HCV RNA negative.

11. Hematology:

1. Absolute neutrophil count greater than 1000/mm3 without the support of
filgrastim.

2. WBC (> 3000/mm(3)).

3. Platelet count greater than 100,000/mm(3).

4. Hemoglobin greater than 8.0 g/dl.

12. Chemistry:

1. Serum ALT/AST less or equal to 2.5 times the upper limit of normal.

2. Serum creatinine less than or equal to 1.6 mg/dl.

3. Total bilirubin less than or equal to 1.5 mg/dl, except in patients with
Gilbert s Syndrome who must have a total bilirubin less than 3.0 mg/dl.

13. More than four weeks must have elapsed since any prior systemic therapy at the
time the patient receives the preparative regimen, and patients toxicities must
have recovered to a grade 1 or less (except for toxicities such as alopecia or
vitiligo).

14. Six weeks must have elapsed since any prior anti-CTLA4 antibody therapy to allow
antibody levels to decline.

15. Patients who have previously received any anti-CTLA4 antibody and have documented
GI toxicity must have a normal colonoscopy with normal colonic biopsies.

EXCLUSION CRITERIA:

1. Previous treatment with IL-12.

2. Women of child-bearing potential who are pregnant or breastfeeding because of the
potentially dangerous effects of the preparative chemotherapy on the fetus or infant.

3. Active systemic infections, coagulation disorders or other major medical illnesses of
the cardiovascular, respiratory or immune system, myocardial infarction, cardiac
arrhythmias, obstructive or restrictive pulmonary disease.

4. Any form of primary immunodeficiency (such as Severe Combined Immunodeficiency
Disease).

5. Concurrent opportunistic infections (The experimental treatment being evaluated in
this protocol depends on an intact immune system. Patients who have decreased immune
competence may be less responsive to the experimental treatment and more susceptible
to its toxicities).

6. Concurrent systemic steroid therapy.

7. History of severe immediate hypersensitivity reaction to any of the agents used in
this study.

8. History of coronary revascularization or ischemic symptoms

9. Any patient known to have an LVEF less than or equal to 45%.

10. Documented LVEF of less than or equal to 45% tested in patients with:

1. History of ischemic heart disease, chest pain, or clinically significant atrial
and/or ventricular arrhythmias including but not limited to: atrial fibrillation,
ventricular tachycardia, second or third degree heart block

2. Age greater than 60 years old

11. Documented FEV1 less than or equal to 60% predicted tested in patients with:

1. A prolonged history of cigarette smoking (20 pk/year of smoking within the past 2
years).

2. Symptoms of respiratory dysfunction