# Enhancing Melanoma TIL Efficacy with Multifactor mRNA-Mediated T Cell Reprogramming

> **NIH NIH R21** · YALE UNIVERSITY · 2024 · $33,048

## Abstract

Refer to Parent Grant

SUMMARY AND SPECIFIC AIMS OF PARENT GRANT (R21CA282629, PI: KATZ). Although T cell mediated 
immune responses are critical for successful immunotherapy, T cells associated with malignant lesions are 
typically dysfunctional and fail to control tumor growth. Treatment with tumor infiltrating lymphocytes (TIL) that 
are isolated, activated, and expanded ex vivo has proven effective in some melanoma patient populations. 
However, many patients do not respond, presumably due to one of several host immune factors. Current 
understanding of TIL mechanism of action suggests both an early robust expansion of tumor-specific effector T 
cells and transfer of less differentiated cells with long-term survival capacity are key to successful therapy. 
Evidence for the former includes the need for high dose exogenous IL-2 at the time of TIL infusion, correlation 
of response with a high frequency of effector T cells, and majority of tumor killing occurring early after the initiation 
of therapy. Evidence for the latter is found in many pre-clinical experiments as well as clinical observations where 
the presence of TILs from the central memory subset in the infusion product correlates with tumor regression. 
Our overall goal is to improve TIL therapeutic efficacy through generation of TIL products with both the transient 
ability to effectively immediately kill tumor cells and the long-term ability to persist and maintain durable antitumor responses. We have developed robust methods to reprogram TILs with mRNA-mediated gene therapy. 
The use of our mRNA approach has the advantages of increased safety, high efficiency, rapid production, tightly 
controlled expression levels, and simultaneous multi-factor reprogramming. In preliminary work, we developed 
a system that increases mRNA lifespan by an order of magnitude. Our single cell analysis of patient TILs preand post-expansion has identified two specific pathways deficient in the expanded TIL product that likely 
contribute to their poor immediate efficacy and absence of memory fate. Both will be augmented by TIL mRNA 
reprogramming. By leveraging our multidisciplinary team and existing infrastructure, the knowledge gained from 
this project will be quickly translatable into clinical trials to examine the efficacy of mRNA reprogramming of TIL 
to improve treatment of patients with advanced melanoma. We will complete the following Specific Aims: 
Aim 1. Determine the ability to enhance post-expansion TIL survival. 
1.1. Increase TIL durability by expression of an anti-apoptotic protein with a novel long-lived (loli) mRNA 
expression system. 
1.2. Augment TIL efficacy by reprogramming with supportive cytokine mRNAs. 
1.3. Analyze efficacy of TILs reprogrammed with survival and cytokine mRNAs in patient derived xenografts. 
Hypothesis: TILs encounter a large bottleneck of elimination upon infusion as they adapt from ex vivo culture 
conditions to the harsh tumor microenvironment. By reprog...

## Key facts

- **NIH application ID:** 10993374
- **Project number:** 3R21CA282629-01S1
- **Recipient organization:** YALE UNIVERSITY
- **Principal Investigator:** MICHAEL E HURWITZ
- **Activity code:** R21 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $33,048
- **Award type:** 3
- **Project period:** 2024-03-01 → 2025-06-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10993374

## Citation

> US National Institutes of Health, RePORTER application 10993374, Enhancing Melanoma TIL Efficacy with Multifactor mRNA-Mediated T Cell Reprogramming (3R21CA282629-01S1). Retrieved via AI Analytics 2026-05-25 from https://api.ai-analytics.org/grant/nih/10993374. Licensed CC0.

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