Trial Information

Phase II Study of Metastatic Melanoma Using Lymphodepleting Conditioning Followed by Infusion of Anti-gp100:154-162 TCR-Gene Engineered Lymphocytes

Background:

- We have engineered human tumor infiltrating lymphocytes (TIL) and peripheral blood
lymphocytes (PBLs) to express a T-cell receptor that recognizes an HLAA 0201 restricted
epitope derived from the gp100 protein.

- We constructed a single retroviral vector that contains both Alpha and Beta chains and
can mediate genetic transfer of this TCR with high efficiency (greater than 30 percent)
without the need to perform any selection.

- In co-cultures with HLA-A*0201 positive melanoma gp100:154-162 TCR transduced T cells
secreted significant amount of IFN-Beta (but no significant secretion was observed in
control co-cultures with cell lines.

- gp100:154-162 TCR transduced T-cells could efficiently kill HLA-A*0201 positive tumors.
There was little or no recognition of normal fibroblasts cells.

- This TCR is over 10 times more reactive with melanoma cells than the MART-1 TCR that
mediated tumor regression in two patients with metastatic melanoma.

Objectives:

Primary objectives:

-Determine if the administration of anti-gp100:154-162 TCR-engineered peripheral blood
lymphocytes (PBL) or tumor infiltrating lymphocytes (TIL) and aldesleukin to patients
following a nonmyeloablative but lymphoid depleting preparative regimen will result in
clinical tumor regression in patients with metastatic melanoma.

Secondary objectives:

- Determine the in vivo survival of TCR gene-engineered cells.

- Determine the toxicity profile of this treatment regimen.

- Determine whether treated patients develop anti-mouse TCR antibody.

Eligibility:

Patients who are HLA-A*0201 positive and 18 years of age or older must have

- metastatic melanoma;

- previously received and have been a non-responder to or recurred after aldesleukin

- normal values for basic laboratory values.

Patients may not have:

- concurrent major medical illnesses;

- any form of primary or secondary immunodeficiency;

- severe hypersensitivity to any of the agents used in this study;

- contraindications for high dose aldesleukin administration.

Design:

- If TIL can be obtained and grown but are non-reactive, patients will be assigned to
receive TIL transduced with the anti-gp100:154-162 TCR retroviral vector. If TIL cannot
be obtained, peripheral blood mononuclear cells (PBMC) will be obtained by
leukapheresis (approximately 5 times 10(9) cells) and cultured in the presence of
anti-CD3 (OKT3) and aldesleukin and transduced with the antigp100:154-162 TCR
retroviral vector. If TIL cells are reactive to autologous tumor or major
histocompatibility complex (MHC)-matched tumor cells or PBL cannot be grown, patients
will not be treated on this protocol.

- Transduction is initiated by exposure of approximately 10(8) to 5 times 10(8) cells to
supernatant containing the anti-gp100:154-162 TCR retroviral vector. These transduced
cells will be expanded and tested for their anti-tumor activity.

- Once engineered lymphocytes are demonstrated to be biologically active according to the
strict criteria outlined in the Certificate of Analysis, patients will receive a
nonmyeloablative but lymphocyte depleting preparative regimen consisting of
cyclophosphamide and fludarabine followed by intravenous infusion of ex vivo tumor
reactive, TCR gene-transduced PBMC plus IV aldesleukin (720,000 IU/kg q8h for a maximum
of 15 doses).

- Patients will undergo complete evaluation of tumor with physical examination, CT
(computed tomography) of the chest, abdomen and pelvis and clinical laboratory
evaluation four to six weeks after treatment and then monthly for approximately 3 to 4
months or until off study criteria are met.

- The study will be conducted using a phase II optimal design where initially 21
evaluable patients will be enrolled into each of two cohorts. If 0 or 1 of the 21
patients per cohort experiences a clinical response, then no further patients will be
enrolled but if 2 or more of the first 21 evaluable patients enrolled in that cohort
have a clinical response, then accrual to that cohort will continue until a total of 41
evaluable patients have been enrolled in that cohort.

- The objective will be to determine in two cohorts if the combination of high dose
aldesleukin, lymphocyte depleting chemotherapy, and anti-gp100:154-162 TCR-gene
engineered lymphocytes (TIL and PBL) is able to be associated with a clinical response
rate that can rule out 5 percent (p0=0.05) in favor of a modest 20 percent PR (partial
response) plus CR (complete response) rate (p1=0.20).