Lifileucel With Reduced Dose Fludarabine/Cyclophosphamide Lymphodepletion and Interleukin-2 for the Treatment of Patients With Unresectable or Metastatic Melanoma

Reduced Dose Fludarabine/Cyclophosphamide Lymphodepletion Before Tumor-Infiltrating Lymphocyte Therapy With Lifileucel in Metastatic Melanoma

This phase II trial tests how well lifileucel, with reduce dose fludarabine and cyclophosphamide for lymphodepletion and interleukin-2, work for treating patients with melanoma that cannot be removed by surgery (unresectable) or that has spread from where it first started (primary site) to other places in the body (metastatic).Lifileucel is made up of specialized immune cells called lymphocytes or T cells that are taken from a patient's tumor, grown in a manufacturing facility and infused back into the preconditioned patient to attack the tumor. Giving Lifileucel with a reduced dose of fludarabine and cyclophosphamide for lymphodepletion and interleukin -2 is being studied in patients with unresectable or metastatic melanoma.

Study Overview

• Status: Completed
• Conditions: Metastatic Melanoma; Unresectable Melanoma; Pathologic Stage IIIC Cutaneous Melanoma AJCC v8; Clinical Stage IV Cutaneous Melanoma AJCC v8
• Intervention / Treatment: Procedure: Magnetic Resonance Imaging; Drug: Cyclophosphamide; Drug: Fludarabine; Procedure: Multigated Acquisition Scan; Biological: Interleukin-2; Procedure: Echocardiography; Procedure: Biospecimen Collection; Biological: Lifileucel; Procedure: Tumor Resection

Detailed Description

PRIMARY OBJECTIVE:

I. The percentage of total TIL clones as measured by the T-cell receptor (TCR) population shared between the tumor infiltrating lymphocyte (TIL) product and peripheral blood mononuclear cells (PBMC).

SECONDARY OBJECTIVES:

I. To evaluate the efficacy parameters of lifileucel (LN-144) in combination with a reduced dose lymphodepletion in patients with unresectable or metastatic melanoma by assessing objective response rate (ORR), progression-free survival (PFS), and overall survival (OS).

II. To characterize the safety profile of lifileucel (LN-144) in combination with a reduced dose lymphodepletion regimen in patients with unresectable or metastatic melanoma.

EXPLORATORY OBJECTIVE:

I. Blood and tumor samples will be banked for future correlative analyses, including flow cytometry, next generation sequencing, immunogenomics and RNA sequencing to characterize the immunome and microenvironment.

OUTLINE:

Patients undergo tumor resection surgery. After the lifileucel is manufactured (approximately 4 weeks later), patients receive cyclophosphamide intravenously (IV) on days -4 to -2 and fludarabine IV on days -4 to -1. Patients then receive lifileucel IV on day 0. Patients also receive up to 6 doses of intraleukin-2 IV.

After completion of study treatment, patients are followed up at day 28, 42, 84, 126, 180, 365, month 18, and month 24.

• Study Type: Interventional
• Enrollment (Actual): 11
• Phase: Phase 2

Contacts and Locations

Study Locations

• United States
  • Kansas
    • Kansas City, Kansas, United States, 66160
      • University of Kansas Cancer Center

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: Adult; Older Adult
• Accepts Healthy Volunteers: No

Description

Inclusion Criteria:

• Males and females age ≥ 18 years Enrollment of patients ≥ 70 years of age may be allowed at principal investigator discretion.
• Eastern Cooperative Oncology Group (ECOG) performance status 0 - 1

• At least one measurable target lesion, as defined by Response Evaluation Criteria in Solid Tumors (RECIST) version (v) 1.1

  • Lesions in previously irradiated areas (or other local therapy) should not be selected as target lesions, unless treatment was ≥ 3 months prior to Screening, and there has been demonstrated disease progression in that lesion
• Women of childbearing potential must have a negative serum pregnancy test 48 hours prior to initiating treatment
• Patients with unresectable or metastatic melanoma (stage IIIc or stage IV)
• Patients must have progressed following 1-3 prior systemic therapy including a programmed cell death protein-1 (PD-1) blocking antibody; and if proto-oncogene B-Raf (BRAF) V600 mutation positive, a BRAF inhibitor or BRAF inhibitor in combination mitogen-activated extracellular signal-regulated kinase (MEK) inhibitor
• At least one resectable lesion (or aggregate of lesions resected) of a minimum 1.5 cm in diameter post-resection to generate TIL; surgical removal with minimal morbidity (defined as any procedure for which expected hospitalization is ≤ 3 days)
• Adequate hematologic and organ function

• Patients must have recovered from all prior therapy-related adverse events (AEs) to ≤ grade 1 (per Common Terminology Criteria for Adverse Events [CTCAE] version [v] 5.0), except for alopecia or vitiligo, prior to enrollment (tumor resection)

  • Patients with documented ≥ grade 2 diarrhea or colitis because of previous treatment with immune checkpoint inhibitor(s) must have been asymptomatic for at least 6 months and/or had a normal colonoscopy post-immune checkpoint inhibitor treatment, by visual assessment, prior to tumor resection
• Patients with immunotherapy-related endocrinopathies stable for at least 6 weeks (eg, hypothyroidism), and controlled with hormonal replacement (non-corticosteroids), are allowed

• Patients must have a washout period of ≥ 28 days from prior anticancer therapy(ies) to the start of the planned reduced dose lymphodepletion (RDL) preconditioning regimen:

  • Targeted therapy: MEK/BRAF or other targeted agents
  • Chemotherapy
  • Immunotherapy: anti-CTLA-4/anti-PD-1, other monoclonal antibodies (mAb), or vaccine
  • Palliative radiation therapy is permitted so long as it does not involve lesions being selected for TIL, or as target or non-target lesions. Washout is not required if all related toxicities have resolved to ≤ grade 1 as per CTCAE v 5.0
• Women of child-bearing potential and men with partners of child-bearing potential must agree to practice sexual abstinence or to use the forms of contraception listed in Child-Bearing Potential/Pregnancy section for the duration of study participation and for 12 months following the last dose of IL-2 or until the first dose of the next anti-cancer therapy, whichever occurs first

Exclusion Criteria:

• Current or anticipating use of other anti-neoplastic or investigational agents while participating in this study
• Is pregnant or breastfeeding
• Patients who have active medical illness(es) that would pose increased risk for study participation, including active systemic infections requiring systemic antibiotics, coagulation disorders, or other active major medical illnesses of the cardiovascular, respiratory, or immune system
• Patients who have been shown to be BRAF mutation positive (V600), but have not received prior systemic therapy with a BRAF inhibitor alone or a BRAF inhibitor in combination with a MEK inhibitor
• Patients who have received an organ allograft or prior cell transfer therapy
• Patients with melanoma of uveal/ocular origin
• Patients who have a history of hypersensitivity to any component or excipient of Lifileucel or other study drugs
• Patients who have any form of primary immunodeficiency (such as severe combined immunodeficiency disease [SCID] and acquired immunodeficiency syndrome [AIDS])
• Patients who have a left ventricular ejection fraction (LVEF) < 45% or New York Heart Association (NYHA) functional classification class > 1
• Patients who have a documented forced expiratory volume in 1 second (FEV1) of ≤ 60%
• Patients who have had another primary malignancy within the previous 3 years (except for carcinoma in situ of the breast, cervix, or bladder; localized prostate cancer; and non-melanoma skin cancer that has been adequately treated)

• Patients with symptomatic and/or untreated brain metastases (of any size and any number)

  • Other protocol defined inclusion/exclusion criteria could apply

Study Plan

How is the study designed?

Design Details

• Primary Purpose: Treatment
• Allocation: N/A
• Interventional Model: Single Group Assignment
• Masking: None (Open Label)

Number of Arms

1

Arms and Interventions

Participant Group / Arm

Intervention / Treatment

Experimental: Treatment (Lifileucel); Lifileucel (LN-144) is an autologous Tumor Infiltrating Lymphocytes (TIL) cell therapy. A tumor sample is resected from each patient for lifileucel manufacturing. Patients then receive the lifileucel regimen which consists of a reduced dose non-myeloablative lymphodepletion, lifileucel infusion followed by interleukin-2.

Procedure: Magnetic Resonance Imaging; Undergo MRI; Other Names: MRI; Magnetic Resonance; Magnetic Resonance Imaging Scan; Medical Imaging, Magnetic Resonance / Nuclear Magnetic Resonance; MR; MR Imaging; MRI Scan; NMR Imaging; NMRI; Nuclear Magnetic Resonance Imaging; Magnetic resonance imaging (procedure); Drug: Cyclophosphamide; Given IV; Other Names: Cytoxan; CTX; (-)-Cyclophosphamide; 2H-1,3,2-Oxazaphosphorine, 2-[bis(2-chloroethyl)amino]tetrahydro-, 2-oxide, monohydrate; Carloxan; Ciclofosfamida; Ciclofosfamide; Cicloxal; Clafen; Claphene; CP monohydrate; CYCLO-cell; Cycloblastin; Cycloblastine; Cyclophospham; Cyclophosphamid monohydrate; Cyclophosphamide Monohydrate; Cyclophosphamidum; Cyclophosphan; Cyclophosphane; Cyclophosphanum; Cyclostin; Cyclostine; Cytophosphan; Cytophosphane; Fosfaseron; Genoxal; Genuxal; Ledoxina; Mitoxan; Neosar; Revimmune; Syklofosfamid; WR- 138719; Asta B 518; B-518; WR-138719; Drug: Fludarabine; Given IV; Other Names: Fluradosa; Procedure: Multigated Acquisition Scan; Undergo MUGA scan; Other Names: Blood Pool Scan; Equilibrium Radionuclide Angiography; Gated Blood Pool Imaging; MUGA; Radionuclide Ventriculography; RNVG; SYMA Scanning; Synchronized Multigated Acquisition Scanning; MUGA Scan; Multi-Gated Acquisition Scan; Radionuclide Ventriculogram Scan; Gated Heart Pool Scan; Biological: Interleukin-2; Given IV; Other Names: IL-2; IL2; Ro-236019; TCGF; Interleukin II; Lymphocyte Mitogenic Factor; Mitogenic Factor; T-Cell Growth Factor; Thymocyte Stimulating Factor; TSF; Interleukin 2; Epidermal Thymocyte Activating Factor; ETAF; IL2 Protein; Interleukin 2 Precursor; T Cell Growth Factor; hIL-2; Procedure: Echocardiography; Undergo echocardiography; Other Names: EC; Procedure: Biospecimen Collection; A tumor sample is resected from each patient and cultured ex vivo to expand the population of tumor infiltrating lymphocytes. After lymphodepletion, patients are infused with Lifileucel followed by IL-2; Other Names: Biological Sample Collection; Biospecimen Collected; Specimen Collection; Undergo blood sample collection; Biological: Lifileucel; Given IV; Other Names: LN-144; Autologous TIL LN-144; Autologous Tumor Infiltrating Lymphocytes LN-144; Contego; Lifileuecel; Procedure: Tumor Resection; Undergo tumor resection

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Percentage of total T-cell receptor (TCR) populations shared between the TIL product and peripheral blood mononuclear cells (PBMCs)	The TCR population is calculated as the frequencies of unique CDR3 sequences as measured by HTBIvc assay. Will be summarized with respect to mean and standard deviation. The percentage changes will also be presented with respect to visual diagrams for each person, e.g., waterfall plot	At day + 42

Secondary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Safety: Incidence of treatment emergent adverse events	Defined as all grades and any attribution until day 28 (EOT visit) after infusion, grade 3-4 of any attribution from day 28 to 6 months, only grade 3-4 attributed to study drugs after 6 months evaluated by Common Terminology Criteria in Adverse Events (CTCAE). Will be presented as percentage with 95% confidence intervals and tabulated by type and grade.	From start of treatment to 6 months after TIL
Safety: Incidence of serious adverse events	Defined as all grades and any attribution until day 28 (EOT visit) after infusion, grade 3-4 of any attribution from day 28 to 6 months, only grade 3-4 attributed to study drugs after 6 months evaluated by CTCAE. Will be presented as percentage with 95% confidence intervals and tabulated by type and grade.	From start of treatment to 6 months after TIL
Transfusion requirements	Defined as all grades and any attribution until day 28 (EOT visit) after infusion, grade 3-4 of any attribution from day 28 to 6 months, only grade 3-4 attributed to study drugs after 6 months evaluated by CTCAE. Will be presented as percentage with 95% confidence intervals and will be summarized with respect to mean and standard deviation.	From start of treatment to 6 months after TIL
Length of hospital stay	Defined as all grades and any attribution until day 28 (EOT visit) after infusion, grade 3-4 of any attribution from day 28 to 6 months, only grade 3-4 attributed to study drugs after 6 months evaluated by CTCAE. Will be presented as percentage with 95% confidence intervals and will be summarized with respect to mean and standard deviation.	From start of treatment to 6 months after TIL
Overall response rate	Measured by Response Evaluation Criteria in Solid Tumor (RECIST) 1.1. Will be presented as percentage with 95% confidence internal using Clopper-Pearson Exact method.	From start of treatment until the end of treatment, up to 2 years
Progression free survival	Will be summarized with Kaplan-Meir curves and estimation of medians with 95% confidence intervals.	From start of treatment until objective tumor progression or death, up to 2 years
Overall survival	Will be summarized with Kaplan-Meir curves and estimation of medians with 95% confidence intervals.	From start of treatment to death due to any cause, up to 2 years

Collaborators and Investigators

• Sponsor: University of Kansas Medical Center
• Collaborators: National Cancer Institute (NCI); Iovance Biotherapeutics, Inc.
• Investigators: Principal Investigator: Muhammad Umair Mushtaq, University of Kansas

Publications and helpful links

General Publications

• Mushtaq MU, Abdelhakim H, Selby L, Shahzad M, Abhyankar SH, McGuirk JP, Doolittle GC. Reduced dose fludarabine and cyclophosphamide lymphodepletion before tumor-infiltrating lymphocyte therapy in melanoma. Future Oncol. 2025 Jun;21(13):1631-1637. doi: 10.1080/14796694.2025.2498842. Epub 2025 May 9.

Study record dates

Study Major Dates

• Study Start (Actual): December 21, 2023
• Primary Completion (Actual): July 10, 2025
• Study Completion (Actual): July 10, 2025

Study Registration Dates

• First Submitted: November 17, 2023
• First Submitted That Met QC Criteria: November 27, 2023
• First Posted (Actual): November 30, 2023

Study Record Updates

• Last Update Posted (Actual): April 30, 2026
• Last Update Submitted That Met QC Criteria: April 25, 2026
• Last Verified: April 1, 2026

More Information

Terms related to this study

• Additional Relevant MeSH Terms: Neoplasms by Site; Neoplasms; Neoplasms by Histologic Type; Skin Diseases; Neuroectodermal Tumors; Neoplasms, Germ Cell and Embryonal; Neoplasms, Nerve Tissue; Neuroendocrine Tumors; Nevi and Melanomas; Skin Neoplasms; Skin and Connective Tissue Diseases; Melanoma; Peptides; Amino Acids, Peptides, and Proteins; Proteins; Organic Chemicals; Investigative Techniques; Clinical Laboratory Techniques; Diagnostic Techniques and Procedures; Diagnosis; Surgical Procedures, Operative; Hydrocarbons; Biological Factors; Chemistry Techniques, Analytical; Spectrum Analysis; Phosphoramide Mustards; Nitrogen Mustard Compounds; Mustard Compounds; Hydrocarbons, Halogenated; Phosphoramides; Organophosphorus Compounds; Intercellular Signaling Peptides and Proteins; Urologic Surgical Procedures; Urogenital Surgical Procedures; Cytokines; Interleukins; Lymphokines; Cyclophosphamide; Interleukin-2; Specimen Handling; Magnetic Resonance Spectroscopy; fludarabine; Transurethral Resection of Bladder; lifileucel; 4-toluenesulfonyl fluoride
• Other Study ID Numbers: STUDY00150697 (Other Identifier: University of Kansas Cancer Center); P30CA168524 (U.S. NIH Grant/Contract); NCI-2023-09309 (Registry Identifier: CTRP (Clinical Trial Reporting Program)); IIT-2022-LDTIL

Drug and device information, study documents

• Studies a U.S. FDA-regulated drug product: Yes
• Studies a U.S. FDA-regulated device product: No
• product manufactured in and exported from the U.S.: No