Administering Peripheral Blood Lymphocytes Transduced With a Murine T-Cell Receptor Recognizing the G12D Variant of Mutated RAS in HLA-A*11:01 Patients

A Phase I/II Study Administering Peripheral Blood Lymphocytes Transduced With a Murine T-Cell Receptor Recognizing the G12D Variant of Mutated RAS in HLA-A*11:01 Patients

Background:

A new cancer therapy takes white blood cells from a person, grows them in a lab, genetically changes them, then gives them back to the person. Researchers think this may help attack tumors in people with certain cancers. It is called gene transfer using anti-Kirsten rat sarcoma virus (KRAS) Glycine(G) to Aspartic Acid(D) substitution at codon 12(G12D) murine T-cell receptor (mTCR) cells.

Objective:

To see if anti-KRAS G12D mTCR cells are safe and cause tumors to shrink.

Eligibility:

Adults ages 18-72 who have cancer with a molecule on the tumors that can be recognized by the study cells

Design:

Participants will be screened with medical history, physical exam, scans, photography, and heart, lung, and lab tests.

An intravenous (IV) catheter will be placed in a large vein in the chest.

Participants will have leukapheresis. Blood will be removed through a needle in an arm. A machine will divide the blood and collect white blood cells. The rest of the blood will be returned to the participant through a needle in the other arm.

A few weeks later, participants will have a hospital stay. They will:

• Get 2 chemotherapy medicines by IV over 5 days.
• Get the changed cells through the catheter. Get up to 9 doses of medicine to help the cells. They may get a shot to stimulate blood cells.
• Recover in the hospital for up to 3 weeks. They will provide blood samples.

Participants will take an antibiotic for at least 6 months.

Participants will have several follow-up visits over 2 years. They will repeat most of the screening tests and may have leukapheresis.

Participants blood will be collected for several years.

Study Overview

• Status: Terminated
• Conditions: Gastric Cancer; Pancreatic Cancer; Rectal Cancer; Colon Cancer; Gastrointestinal Cancer
• Intervention / Treatment: Drug: Cyclophosphamide; Drug: Fludarabine; Drug: Aldesleukin; Biological: anti-Kirsten rat sarcoma virus (KRAS) Glycine(G) to Aspartic Acid(D) substitution at codon 12 peripheral blood lymphocytes (PBL); Diagnostic test: EKG; Diagnostic test: CT; Diagnostic test: MRI; Diagnostic test: PET; Diagnostic test: Chest x-ray; Other: Photography

Detailed Description

Background:

• We generated an human leukocyte antigen (HLA)-A11:01-restricted murine T-cell receptor (mTCR) that specifically recognizes the Glycine(G) to Aspartic Acid(D) substitution at codon 12 (G12D)-mutated variant of Kirsten rat sarcoma virus (KRAS) (and other RAS family genes), expressed by many human cancers and constructed a single retroviral vector that contains alpha and beta chains that confer recognition of this antigen when transduced into peripheral blood lymphocytes (PBL).
• In co-cultures with HLA-A11:01+ target cells expressing this mutated oncogene, mTCR transduced T-cells lyse target cells and secrete interferon (IFN)-gamma with high specificity.

Objectives:

-Primary objectives:

• Phase I: Determine the safety of administering PBL transduced with anti-KRAS G12D mTCR in concert with preparative lymphodepletion and high-dose interleukin-2 (IL-2; aldesleukin).
• Phase II: Determine if anti-KRAS G12D mTCR-transduced PBL can mediate the regression of tumors harboring the rat sarcoma (RAS) G12D mutation.

Eligibility:

• Patients must be/have:

  • Age greater than or equal to 18 years and less than or equal to 72 years
  • HLA-A*11:01 positive
  • Metastatic or unresectable RAS G12D-expressing cancer which has progressed after standard therapy (if available).

• Patients may not have:

  • Allergies or hypersensitivities to high dose aldesleukin, cyclophosphamide, or fludarabine.

Design:

• This is a phase I/II, single center study of PBL transduced with anti-KRAS G12D mTCR in HLA-A*11:01 positive patients with advanced solid tumors expressing G12D mutated RAS.
• PBMC obtained by leukapheresis will be cultured in the presence of anti-cluster of differentiation 3 (CD3) Ortho-Kung T-cell 3 (OKT3) and aldesleukin in order to stimulate T-cell growth.
• Transduction is initiated by exposure of these cells to retroviral vector supernatant containing replication-incompetent virus encoding the anti-KRAS G12D mTCR.
• All patients will receive a non-myeloablative, lymphodepleting preparative regimen of cyclophosphamide and fludarabine.
• On Day 0, patients will receive their PBL transduced with the anti-KRAS G12D mTCR and will then begin high dose aldesleukin.
• A complete evaluation of lesions will be conducted approximately 6 weeks (plus-minus 2 weeks) after treatment.
• The study will be conducted using a phase I/II Simon minimax design, with two separate cohorts for the Phase II component: Cohort 2a, patients with RAS G12D pancreatic cancer, and Cohort 2b, patients with RAS G12D non-pancreatic cancer.
• A total of up to 70 patients may be required; approximately 24 patients in the Phase I portion of the study and 46 (21, plus an allowance of up to 2 non-evaluable per Phase II cohort) patients in the Phase II portion of the study.

• Study Type: Interventional
• Enrollment (Actual): 5
• Phase: Phase 2; Phase 1

Contacts and Locations

Study Locations

• United States
  • Maryland
    • Bethesda, Maryland, United States, 20892
      • National Institutes of Health Clinical Center

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: 18 years to 70 years (Adult, Older Adult)
• Accepts Healthy Volunteers: No

Description

-INCLUSION CRITERIA:

1. Measurable (per Response Evaluation Criteria in Solid Tumors v1.1 criteria), metastatic, or unresectable malignancy expressing G12D mutated KRAS as assessed by one of the following methods: Reverse Transcription Polymerase Chain Reaction (RT-PCR) on tumor tissue, tumor deoxyribonucleic acid (DNA) sequencing, or any other Clinical Laboratory Improvement Amendments (CLIA)-certified laboratory test on resected tissue. Patients shown to have tumors expressing Glycine(G) to Aspartic Acid(D) substitution at codon 12(G12D) mutated neuroblastoma rat sarcoma (NRAS) and Harvey rat sarcoma viral oncogene homolog (HRAS) will also be eligible as these oncogenes share complete amino acid homology with G12D mutated KRAS for their first 80 N-terminal amino acids, completely encompassing the target epitope.
2. Patients must be human leukocyte antigens (HLA) -A*11:01 positive as confirmed by the National Institutes of Health (NIH) Department of Transfusion Medicine.
3. Confirmation of the diagnosis of cancer by the National Cancer Institute (NCI) Laboratory of Pathology.

4. Patients must have:

   -previously received standard systemic therapy for their advanced cancer and have been either non-responders or have recurred, specifically:

   • Patients with metastatic colorectal cancer must have had at least two systemic chemotherapy regimens that include 5-Fluorouracil (5-FU), leucovorin, bevacizumab, oxaliplatin, and irinotecan (or similar agents), or have contraindications to receiving those medications.
   • Patients with pancreatic cancer must have received gemcitabine, 5FU, and oxaliplatin (or similar agents), or have contraindications to receiving those medications.
   • Patients with non-small cell lung cancer (NSCLC) must have had appropriate targeted therapy as indicated by abnormalities in anaplastic lymphoma kinase (ALK), estimated glomerular filtration rate (EGFR), or expression of programmed death-ligand 1 (PD-L1). Other patients must have had platinum-based chemotherapy.
   • Patients with ovarian cancer or prostate cancer must have had approved first-line chemotherapy.

   OR

   -declined standard treatment.

5. Patients with 3 or fewer brain metastases that are < 1 cm in diameter and asymptomatic are eligible. Lesions that have been treated with stereotactic radiosurgery must be clinically stable for one month after treatment for the patient to be eligible. Patients with surgically resected brain metastases are eligible.
6. Age greater than or equal to 18 years and less than or equal to 72 years.
7. Clinical performance status of Eastern Cooperative Oncology Group (ECOG) 0 or 1
8. Patients must be willing to practice birth control from the time of enrollment on this study and for 12 months after the last dose of combined chemotherapy for women and for four months after treatment for men.

9. Women of child-bearing potential must be willing to undergo a pregnancy test prior to the start of treatment because of the potentially dangerous effects of the treatment on the fetus.

   NOTE: Certain malignancies may secrete hormones that produce false positive pregnancy tests. Serial blood testing (e.g. Human chorionic gonadotropin (hCG) measurements) and/ or ultrasound may be performed for clarification.

10. 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 may have decreased immune-competence and thus may be less responsive to the experimental

    treatment and more susceptible to its toxicities.)

    -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 hepatitis C virus ribonucleic acid (HCV RNA) negative

11. Hematology

    • Absolute neutrophil count (ANC) > 1000/mm^3 without the support of filgrastim
    • White blood cell (WBC) greater than or equal to 2500/mm^3
    • Platelet count greater than or equal to 80,000/mm^3
    • Hemoglobin > 8.0 g/dL. Subjects may be transfused to reach this cut-off.

12. Chemistry

    • Serum alanine Aminotransferase (ALT)/aspartate aminotransferase (AST) less than or equal to 5.0 x ULN
    • 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 < 3.0 mg/dL.

13. Patients must have completed any prior systemic therapy at the time of enrollment.

    Note: Patients may have undergone minor surgical procedures or limited field radiotherapy within the four weeks prior to enrollment, as long as related major organ toxicities have recovered to less than or equal to grade 1.

14. Ability of subject to understand and the willingness to sign a written informed consent document.
15. Willing to sign a durable power of attorney.
16. Subjects must be co-enrolled on the protocol 03C0277.

EXCLUSION CRITERIA:

1. Women of child-bearing potential who are pregnant or breastfeeding because of the potentially dangerous effects of the treatment on the fetus or infant.
2. Concurrent systemic steroid therapy.
3. Active systemic infections requiring anti-infective treatment, coagulation disorders, or any other active or uncompensated major medical illnesses.
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. History of severe immediate hypersensitivity reaction to cyclophosphamide, fludarabine, or aldesleukin.
7. History of coronary revascularization or ischemic symptoms.
8. For select patients with a clinical history prompting cardiac evaluation: last known left ventricular ejection fraction (LVEF) less than or equal to 45%.

I. For select patients with a clinical history prompting pulmonary evaluation: known forced expiratory volume at one second (FEV1) less than or equal to 50%.

j. Patients who are receiving any other investigational agents.

Study Plan

How is the study designed?

Design Details

• Primary Purpose: Treatment
• Allocation: Non-Randomized
• Interventional Model: Sequential Assignment
• Masking: None (Open Label)

Number of Arms

2

Arms and Interventions

Participant Group / Arm	Intervention / Treatment
Experimental: Arm 1/Phase I Non-myeloablative, Lymphodepleting Preparative Regimen; Cohort 1, Arm 1. Participants enrolled with measurable, metastatic, or unresectable malignancy expressing G12D mutated KRAS. Non-myeloablative, lymphodepleting preparative regimen of cyclophosphamide and fludarabine + escalating doses of anti-Kirsten rat sarcoma virus (KRAS) Glycine(G) to Aspartic Acid(D) substitution at codon 12(G12D) murine T-cell receptor (mTCR) peripheral blood lymphocytes (PBL) + highdose aldesleukin.	Drug: Cyclophosphamide; Days -7 and -6: Cyclophosphamide 60 mg/kg/day x 2 days intravenous (IV) in 250 mL 5% Dextrose in water (D5W) infused simultaneously with mesna 15 mg/kg/day over 1 hour x 2 days.; Other Names: Cytoxan; Neosar; Drug: Fludarabine; Days -7 to -3: Fludarabine 25 mg/m^2/day intravenous piggyback (IVPB) daily over 30 minutes for 5 days.; Other Names: Fludara; Drug: Aldesleukin; Aldesleukin 720,000 IU/kg intravenous (IV) (based on total body weight) over 15 minutes approximately every 8 hours beginning within 24 hours of cell infusion and continuing for up to 3 days (maximum 9 doses).; Other Names: Interleukin- 2; Biological: anti-Kirsten rat sarcoma virus (KRAS) Glycine(G) to Aspartic Acid(D) substitution at codon 12 peripheral blood lymphocytes (PBL); Day 0: Cells will be infused intravenously on the Patient Care Unit over 20-30 minutes (2-4 days after the last dose of fludarabine).; Diagnostic test: EKG; Baseline within 14 days prior to preparative regimen; Other Names: Electrocardiogram; Diagnostic test: CT; Within 6 weeks and post treatment follow-up; Other Names: Computed tomography; Diagnostic test: MRI; Within 6 weeks and post treatment follow-up; Other Names: Magnetic resonance imaging; Diagnostic test: PET; Within 6 weeks and post treatment follow-up; Other Names: positron emission tomography; Diagnostic test: Chest x-ray; Baseline within 14 days prior to preparative regimen; Other Names: Chest XR; Other: Photography; Within 6 weeks and post treatment follow-up
Experimental: Arm 2/Phase II Non-myeloablative, Lymphodepleting Preparative Regimen; Cohort 2a: Participants with a diagnosis of pancreatic cancer. Cohort 2b: Participants with a diagnosis other than pancreatic cancer. Non-myeloablative, lymphodepleting preparative regimen of cyclophosphamide and fludarabine + maximum tolerated dose (MTD) of anti-Kirsten rat sarcoma virus (KRAS) Glycine(G) to Aspartic Acid(D) substitution at codon 12(G12D) murine T-cell receptor (mTCR) peripheral blood lymphocytes (PBL) + high-dose aldesleukin.	Drug: Cyclophosphamide; Days -7 and -6: Cyclophosphamide 60 mg/kg/day x 2 days intravenous (IV) in 250 mL 5% Dextrose in water (D5W) infused simultaneously with mesna 15 mg/kg/day over 1 hour x 2 days.; Other Names: Cytoxan; Neosar; Drug: Fludarabine; Days -7 to -3: Fludarabine 25 mg/m^2/day intravenous piggyback (IVPB) daily over 30 minutes for 5 days.; Other Names: Fludara; Drug: Aldesleukin; Aldesleukin 720,000 IU/kg intravenous (IV) (based on total body weight) over 15 minutes approximately every 8 hours beginning within 24 hours of cell infusion and continuing for up to 3 days (maximum 9 doses).; Other Names: Interleukin- 2; Biological: anti-Kirsten rat sarcoma virus (KRAS) Glycine(G) to Aspartic Acid(D) substitution at codon 12 peripheral blood lymphocytes (PBL); Day 0: Cells will be infused intravenously on the Patient Care Unit over 20-30 minutes (2-4 days after the last dose of fludarabine).; Diagnostic test: EKG; Baseline within 14 days prior to preparative regimen; Other Names: Electrocardiogram; Diagnostic test: CT; Within 6 weeks and post treatment follow-up; Other Names: Computed tomography; Diagnostic test: MRI; Within 6 weeks and post treatment follow-up; Other Names: Magnetic resonance imaging; Diagnostic test: PET; Within 6 weeks and post treatment follow-up; Other Names: positron emission tomography; Diagnostic test: Chest x-ray; Baseline within 14 days prior to preparative regimen; Other Names: Chest XR; Other: Photography; Within 6 weeks and post treatment follow-up

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Phase I: Number of Grades 1, 2, 3, 4, and/or 5 and Type of Serious and/or Non-serious Toxicities Experienced by Participants at Each Dose Level	All participants will be evaluable for toxicity from the time of their first treatment with cyclophosphamide. Toxicity was assessed by the Common Terminology Criteria for Adverse Events (CTCAE v5.0). A non-serious adverse event is any untoward medical occurrence. A serious adverse event is an adverse event or suspected adverse reaction that results in death, a life-threatening adverse drug experience, hospitalization, disruption of the ability to conduct normal life functions, congenital anomaly/birth defect or important medical events that jeopardize the patient or subject and may require medical or surgical intervention to prevent one of the previous outcomes mentioned. Grade 1 is mild. Grade 2 is moderate. Grade 3 is severe. Grade 4 is life-threatening. Grade 5 is death related to adverse event.	from the start of cyclophosphamide, through the first follow-up evaluation (6 weeks [± 2 weeks]) following administration of the cell product) until off study, whichever comes first, an average of 2.3 months
Phase I: Percentage of Participants Who Experienced a Grade 3, 4 and/or 5 Serious Dose-Limiting Toxicity (DLT) Reported With Type at Each Dose Level	Toxicity was assessed by the Common Terminology Criteria for Adverse Events. A DLT is all grade 3 and greater toxicities related to the cell infusion with exceptions, such as myelosuppression, defined as lymphopenia, neutropenia, decreased hemoglobin and thrombocytopenia due to the non-myeloablative lymphodepleting preparative regimen, expected chemotherapy toxicities, Aldesleukin expected toxicities, immediate hypersensitivity reactions occurring within 2 hours of cell infusion that are reversible to a grade 2 or less within 24 hours of cell administration with standard therapy, Grade 3 fever, Grade 3 metabolic laboratory abnormalities without significant clinical sequela that resolve to grade 2 within 7 days, Grade 3 autoimmune toxicity that resolves to grade 2 or less within 10 days unless immunosuppression is required, and events that are clearly related to the participants disease. Grade 3 is severe. Grade 4 is life-threatening. Grade 5 is death related to adverse event.	At the time of cell infusion and end two weeks after cell infusion, an average of one month
Phase II: Percentage of Participants With a Clinical Response (Complete Response (CR) + Partial Response (PR) Reported With 80% Confidence Interval	Percentage of participants who have a clinical response (PR+CR) to treatment (objective tumor regression). Response was assessed by the Response Evaluation Criteria in Solid Tumors (RECIST) (version 1.1). CR is a disappearance of all target lesions. PR is at least a 20% increase in the sum of the diameters of target lesions, taking as reference the smallest sum on study (this includes the baseline sum if that is the smallest on study).	6 weeks and 12 weeks following administration of the cell product, then every 3 months x3, then every 6 months x 2 years, then per principal investigator (PI) discretion
Phase II: Percentage of Participants With a Clinical Response (Complete Response (CR) + Partial Response (PR) Reported With 95% Confidence Interval	Percentage of participants who have a clinical response (PR+CR) to treatment (objective tumor regression). Response was assessed by the Response Evaluation Criteria in Solid Tumors (RECIST) (version 1.1). CR is a disappearance of all target lesions. PR is at least a 20% increase in the sum of the diameters of target lesions, taking as reference the smallest sum on study (this includes the baseline sum if that is the smallest on study).	6 weeks and 12 weeks following administration of the cell product, then every 3 months x3, then every 6 months x 2 years, then per principal investigator (PI) discretion

Other Outcome Measures

Outcome Measure	Measure Description	Time Frame
Phase I: Number of Participants With Serious and/or Non-serious Adverse Events Assessed by the Common Terminology Criteria for Adverse Events (CTCAE v5.0).	Adverse events were assessed by the Common Terminology Criteria for Adverse Events (CTCAE v5.0). A non-serious adverse event is any untoward medical occurrence. A serious adverse event is an adverse event or suspected adverse reaction that results in death, a life-threatening adverse drug experience, hospitalization, disruption of the ability to conduct normal life functions, congenital anomaly/birth defect or important medical events that jeopardize the patient or subject and may require medical or surgical intervention to prevent one of the previous outcomes mentioned.	from the start of cyclophosphamide, through the first follow-up evaluation (6 weeks [± 2 weeks]) following administration of the cell product) until off study, whichever comes first, an average of 2.3 months

Collaborators and Investigators

• Sponsor: National Cancer Institute (NCI)
• Investigators: Principal Investigator: James C Yang, M.D., National Cancer Institute (NCI)

Publications and helpful links

General Publications

• Abrams SI, Khleif SN, Bergmann-Leitner ES, Kantor JA, Chung Y, Hamilton JM, Schlom J. Generation of stable CD4+ and CD8+ T cell lines from patients immunized with ras oncogene-derived peptides reflecting codon 12 mutations. Cell Immunol. 1997 Dec 15;182(2):137-51. doi: 10.1006/cimm.1997.1224.
• Davis JL, Theoret MR, Zheng Z, Lamers CH, Rosenberg SA, Morgan RA. Development of human anti-murine T-cell receptor antibodies in both responding and nonresponding patients enrolled in TCR gene therapy trials. Clin Cancer Res. 2010 Dec 1;16(23):5852-61. doi: 10.1158/1078-0432.CCR-10-1280.
• Wang QJ, Yu Z, Griffith K, Hanada K, Restifo NP, Yang JC. Identification of T-cell Receptors Targeting KRAS-Mutated Human Tumors. Cancer Immunol Res. 2016 Mar;4(3):204-14. doi: 10.1158/2326-6066.CIR-15-0188. Epub 2015 Dec 23.

Helpful Links

• NIH Clinical Center Detailed Web Page

Study record dates

Study Major Dates

• Study Start (Actual): May 16, 2019
• Primary Completion (Actual): August 23, 2022
• Study Completion (Actual): August 23, 2022

Study Registration Dates

• First Submitted: November 16, 2018
• First Submitted That Met QC Criteria: November 16, 2018
• First Posted (Actual): November 19, 2018

Study Record Updates

• Last Update Posted (Actual): May 19, 2026
• Last Update Submitted That Met QC Criteria: May 5, 2026
• Last Verified: May 1, 2026

More Information

Terms related to this study

• Keywords: Cell Therapy; Immunotherapy; KRAS; NRAS; HRAS
• Additional Relevant MeSH Terms: Endocrine System Diseases; Neoplasms by Site; Neoplasms; Intestinal Diseases; Digestive System Neoplasms; Digestive System Diseases; Gastrointestinal Diseases; Stomach Diseases; Colorectal Neoplasms; Intestinal Neoplasms; Rectal Diseases; Endocrine Gland Neoplasms; Pancreatic Diseases; Colonic Diseases; Stomach Neoplasms; Rectal Neoplasms; Colonic Neoplasms; Pancreatic Neoplasms; Gastrointestinal Neoplasms; Peptides; Amino Acids, Peptides, and Proteins; Proteins; Organic Chemicals; Investigative Techniques; Diagnostic Techniques and Procedures; Diagnosis; Hydrocarbons; Biological Factors; Tomography; Phosphoramide Mustards; Nitrogen Mustard Compounds; Mustard Compounds; Hydrocarbons, Halogenated; Phosphoramides; Organophosphorus Compounds; Intercellular Signaling Peptides and Proteins; Diagnostic Techniques, Cardiovascular; Cytokines; Interleukins; Lymphokines; Radiography; Heart Function Tests; Electrodiagnosis; Image Interpretation, Computer-Assisted; Radiographic Image Enhancement; Image Enhancement; Tomography, X-Ray; Tomography, Emission-Computed; Radionuclide Imaging; Diagnostic Techniques, Radioisotope; Cyclophosphamide; Interleukin-2; Magnetic Resonance Imaging; fludarabine; aldesleukin; fludarabine phosphate; Electrocardiography; Tomography, X-Ray Computed; Positron-Emission Tomography; Photography; Diagnostic Imaging
• Other Study ID Numbers: 190017; 19-C-0017

Plan for Individual participant data (IPD)

• Plan to Share Individual Participant Data (IPD)?: YES
• IPD Plan Description: All individual participant data (IPD) recorded in the medical record will be shared with intramural investigators upon request. All collected IPD will be shared with collaborators under the terms of collaborative agreements.
• IPD Sharing Time Frame: Clinical data will be available during the study and indefinitely.
• IPD Sharing Access Criteria: Clinical data will be made available via subscription to Biomedical Translational Research Information System (BTRIS) and with the permission of the study principal investigator (PI).
• IPD Sharing Supporting Information Type: STUDY_PROTOCOL; SAP; ICF

Drug and device information, study documents

• Studies a U.S. FDA-regulated drug product: Yes
• Studies a U.S. FDA-regulated device product: No