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Phase I/II Study Using a Non-Myeloablative Lymphocyte Depleting Regimen of Chemotherapy Followed by Infusion of Allogeneic Tumor-Reactive Lymphocyte Cell Line DMF5 in Metastatic Melanoma

Phase 1/Phase 2
18 Years
Not Enrolling
Melanoma, Malignant Melanoma, Melanoma, Experimental

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Trial Information

Phase I/II Study Using a Non-Myeloablative Lymphocyte Depleting Regimen of Chemotherapy Followed by Infusion of Allogeneic Tumor-Reactive Lymphocyte Cell Line DMF5 in Metastatic Melanoma


In previous trials in the Surgery Branch, a 51 percent objective response rate has been
observed in heavily pre-treated patients with metastatic melanoma undergoing adoptive cell
transfer therapy utilizing a non-myeloablative preparative regimen followed by
administration of autologous tumor-reactive lymphocytes and subsequent treatment with
high-dose aldesleukin.

However, in patients with metastatic melanoma undergoing metastasectomy, recovery of
adequate numbers of tumor specific T lymphocytes from surgical specimens is possible in
approximately half of all patients, thus limiting the application of adoptive cell transfer

Murine models performed in the Surgery Branch have demonstrated solid tumor regression in
mice treated with allogeneic tumor specific T cells combined with a preinfusion
lymphodepleting regimen.

We have identified a tumor specific lymphocyte cell line (DMF5) used previously in an
autologous adoptive cell transfer protocol that was associated with an objective clinical
response in that patient.

In subsequent preclinical testing of this lymphocyte population, we have demonstrated high
specificity against HLA-A 0201 positive melanoma cell lines as well as the common shared
melanocyte differentiation antigen MART-1:27-35. We have expanded this lymphocyte population
to provide up to 30 individual allogeneic cell transfers to HLAA 0201 positive patients with
metastatic melanoma.

In this trial we want to test our hypothesis that objective tumor regression can be achieved
with the DMF5 allogeneic T-cell product using a non-myeloablative regimen followed by cell
transfer and high-dose aldesleukin.

It should be emphasized that this protocol is designed to test whether highly melanoma
reactive allogeneic lymphocytes can mediate cancer regression. The DMF5 cell line is a
limited reagent only available for the treatment of up to 30 patients. However, if this
treatment results in cancer regression, it will represent an important step in our
development of an allogeneic T-cell receptor engineered universal effector cell line for the
treatment of patients with cancer.


To evaluate the safety of the administration of the DMF5 allogeneic T-cell product in
patients receiving the non- myeloablative conditioning regimen, and aldesleukin.

To determine whether this allogeneic tumor-specific lymphocyte cell line, hereafter referred
to as DMF5, infused in conjunction with the administration of high-dose aldesleukin may
result in objective clinical tumor regression in eligible HLA-A 0201 positive patients with
metastatic melanoma receiving a non-myeloablative lymphoid depleting preparative regimen.

To determine the in vivo survival of the infused cells following the non-myeloablative
regimen, via analysis of the sequence of the variable region of the T cell receptor or flow
cytometry (FACS).


Patients with metastatic melanoma who are greater than or equal to 18 years of age, HLA-A
0201 positive, do not have suitable autologous tumor reactive TIL cells available, and are
able to tolerate high-dose aldesleukin.


Patients will receive a non-myeloablative lymphocyte depleting preparative regiment
consisting of cyclophosphamide (60 mg/kg/day times 2 days intravenous (IV)) and fludarabine
(25 mg/m2/day IV times 5 days).

Patients will receive intravenous adoptive transfer of the tumor reactive lymphocyte cell
line DMF5 (after its expansion in interleukin-2 and OKT3) followed by high-dose intravenous
(IV) aldesleukin (720,000 IU/kg/dose every 8 hours for up to 15 doses).

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 and then monthly for approximately 3 to 4 months or 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, with three cohorts. Should a single patient
experience a dose limiting toxicity at a particular dose level, three more 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 prior to
starting the phase II portion. If a dose limiting toxicity 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 would proceed to the phase II portion, and
initially, 9 total patients will be administered the therapy at the maximum tolerated dose.
The plan will utilize a Simon two-stage optimal phase II design. If 0 of the 9 patients
experiences a clinical response, then no further patients will be enrolled but if 1 or more
of the first 9 evaluable patients enrolled have a clinical response, then accrual will
continue until a total of 30 evaluable patients have been enrolled.

This design has the ability to distinguish a 5% response rate (p0=0.05) from a 25% response
rate (p1=0.25), with 10% probability of falsely "accepting" the DMF5 cell therapy approach
(alpha=0.10), and 10% probability of incorrectly discarding this strategy as if it were
unacceptably poor (beta=0.10). This design also has 63% probability of stopping early (at 9
patients) if the true response rate is 5%.

Inclusion Criteria


1. Measurable metastatic melanoma that is refractory to standard treatment including
high dose aldesleukin.

2. Unsuitable autologous cells for Institutional Review Board (IRB) approved Surgery
Branch adoptive cell therapy studies.

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

4. Life expectancy of greater than three months.

5. Willing to sign a durable power of attorney.

6. Able to understand and sign the Informed Consent Document.

7. Human leukocyte antigen A (HLA-A) 0201 positive.

8. Willing to practice birth control during treatment and for four months after
receiving the preparative regimen.

9. Clinical performance status of Eastern Cooperative Oncology Group (ECOG) 0 or 1.

10. Hematology:

- Absolute neutrophil count greater than 1000/mm^3 without support of filgrastim.

- WBC greater than 3000/mm^3.

- Hemoglobin greater than 8.0 g/dl.

- Platelet count greater than 100,000/mm^3.

11. Serology:

- Seronegative for human immunodeficiency virus (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.)

- Seronegative for hepatitis B antigen and hepatitis C antibody unless antigen

12. Chemistry:

- Serum alanine aminotransferase (ALT)/aspartate aminotransferase (AST) less than
three times the upper limit of normal.

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

- Total bilirubin less than or equal to 2.0 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 prior Ipilimumab (MDX-010) therapy to allow
antibody levels to decline.

15. Patients who have previously received MDX-010 must have a normal colonoscopy with
normal colonic biopsies.


1. 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.

2. 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.

3. Any form of primary immunodeficiency (such as Severe Combined Immunodeficiency

4. Opportunistic infections (The experimental treatment being evaluated in his 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

5. Symptomatic central nervous system (CNS) lesions (Patients maybe eligible after
treatment of their symptomatic lesions.)

6. 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. Patients with a prolonged (greater than 20 pk/yrs) history of cigarette smoking or
symptoms of respiratory dysfunction with pulmonary function tests (PFT's) indicating
an forced expiratory volume (FEV1) less than 60 percent predicted for age.

10. Patients with a history of clinically significant atrial and/or ventricular
arrhythmias including but not limited to: atrial fibrillation, ventricular
tachycardia, heart block or greater than or equal to age 60 with an left ventricular
ejection fraction (LVEF) of less than 45 percent on cardiac evaluation
(echocardiogram, multi-gated acquisition scan (MUGA), etc.) will be excluded.

11. Positive allo-specific reactivity of the DMF5 cells to the patient's peripheral blood
mononuclear cells (PBMC).

12. Documented penicillin allergy.

Type of Study:


Study Design:

Endpoint Classification: Safety/Efficacy Study, Intervention Model: Single Group Assignment, Masking: Open Label, Primary Purpose: Treatment

Outcome Measure:

Number of Participants With an Objective Clinical Tumor Regression Response According to RECIST Criteria

Outcome Description:

Response is determined by the Response Evaluation Criteria in Solid Tumors (RECIST). Complete response (CR) is the disappearance of all target lesions, partial response (PR) is at least a 30% decrease in the target lesions, progression (PD) is at least a 20% increase in the target lesions or appearance of one or more new lesions, and stable disease is neither sufficient shrinkage to qualify for PR nor sufficient increase to qualify for PD.

Outcome Time Frame:

44 days

Safety Issue:


Principal Investigator

Steven Rosenberg, M.D.

Investigator Role:

Principal Investigator

Investigator Affiliation:

National Cancer Institute, National Institutes of Health


United States: Federal Government

Study ID:




Start Date:

August 2007

Completion Date:

October 2010

Related Keywords:

  • Melanoma
  • Malignant Melanoma
  • Melanoma, Experimental
  • Clinical Response
  • Immunotherapy
  • Cancer
  • Cytokines
  • Adoptive Cell Therapy
  • Melanoma
  • Metastatic Melanoma
  • Melanoma
  • Melanoma, Experimental



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