# A High Throughput Method to Determine the Target of T Cell Receptors

> **NIH NIH F31** · WEILL MEDICAL COLL OF CORNELL UNIV · 2021 · $46,036

## Abstract

Project Summary/Abstract
The immune system has developed to identify and target foreign or mutated intracellular proteins to defend
against cancers, viruses and infections. Intracellular proteins are presented as peptides in major
histocompatibility complexes on the surfaces of cells, which are recognized by T cell receptors (TCR) on CD8+
T cells. TCRs are attractive for cancer therapy because they can specifically target peptide-MHC (pMHC)
antigens found across numerous patients and cancers, known as shared antigens. Shared antigens have
been identified from cancer testis antigens and melanoma antigens. Endogenous TCRs targeting these
antigens have been identified from patient samples. These TCRs have been successful as treatment in
numerous patients when exogenously expressed in autologous T cells and reinfused. While, tumor reactive
TCRs have a great potential for use in therapy, it remains challenging to identify the target of tumor reactive
TCRs. This limits the repertoire of known shared antigens that can be targeted for therapy. Here we propose
a high throughput method to identify the targets of TCRs, thereby increasing the potential of TCR based
therapies and identifying novel shared antigens. To identify the targets of TCRs, a DNA library encoding
peptides will be used to present a large array of representative pMHCs. The peptide libraries are designed to
have variability in positions responsible for the majority of TCR contacts, biasing the libraries to be more
reactive. To determine TCR targets in a functionally relevant manner, relying on the cytotoxicity of T cells,
coculture depletion screens will be performed. Peptides depleted from the screens are considered hits. The
hits will be used to identify peptide binding motifs related to TCR reactivity. From these motifs clinically
relevant, potential targets will be identified. The targets will be validated using single peptide coculture killing
assays and ELISpot for identification of interferon gamma, a marker of T cell reactivity. This technology will be
used to identify clinically relevant targets of TCRs from tumor infiltrating lymphocytes.

## Key facts

- **NIH application ID:** 10214540
- **Project number:** 5F31CA253995-02
- **Recipient organization:** WEILL MEDICAL COLL OF CORNELL UNIV
- **Principal Investigator:** Heather Jones
- **Activity code:** F31 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $46,036
- **Award type:** 5
- **Project period:** 2020-07-02 → 2023-07-01

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10214540

## Citation

> US National Institutes of Health, RePORTER application 10214540, A High Throughput Method to Determine the Target of T Cell Receptors (5F31CA253995-02). Retrieved via AI Analytics 2026-05-22 from https://api.ai-analytics.org/grant/nih/10214540. Licensed CC0.

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