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

> **NIH NIH R21** · YALE UNIVERSITY · 2024 · $185,977

## Abstract

PROJECT SUMMARY
Although T cell mediated immune responses are critical for the success of immunotherapy, those 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 very effective
in some patient populations of melanoma. However, a substantial number of patients do not respond,
presumably due to one of a number of host immune factors. Current understanding of TIL mechanism of action
suggests that 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 a successful therapy. Evidence for the former includes the need
for high dose exogenous IL-2 at the time of TIL infusion, the correlation of response with a high frequency of
effector T cells, and the majority of tumor killing occurring very 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 the generation of TIL products with both the transient ability to effectively
immediately kill tumor cells as well as the long-term ability to persist and maintain durable anti-tumor responses.
To address these challenges 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 have developed a system that increases mRNA lifespan by an order of magnitude. Our single cell
analysis of patient TILs pre- and 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 of these
will be augmented by TIL mRNA reprogramming. In Aim 1 we will develop a method to enhance post-expansion
TIL survival, while in Aim 2 we will improve the production of central memory TILs. We will evaluate the effect
of these improvements in TIL production using paired tumor/TIL sets derived from the melanomas from multiple
patients by studying tumor-mediated TIL activation and tumor lysis in vitro and in pre-clinical humanized mouse
models using single cell analysis and advanced spatial transcriptomics. This application addresses the need to
improve response rates to adoptive cell immunotherapies for melanoma and is designed to be translatable to
clinical trials in the near future.

## Key facts

- **NIH application ID:** 10851011
- **Project number:** 5R21CA282629-02
- **Recipient organization:** YALE UNIVERSITY
- **Principal Investigator:** MICHAEL E HURWITZ
- **Activity code:** R21 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $185,977
- **Award type:** 5
- **Project period:** 2023-07-01 → 2025-06-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10851011

## Citation

> US National Institutes of Health, RePORTER application 10851011, Enhancing Melanoma TIL Efficacy with Multifactor mRNA-Mediated T Cell Reprogramming (5R21CA282629-02). Retrieved via AI Analytics 2026-05-24 from https://api.ai-analytics.org/grant/nih/10851011. Licensed CC0.

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