# Optimizing TCR-CD3 signaling for immunotherapy of cancer

> **NIH NIH R01** · NEW YORK UNIVERSITY SCHOOL OF MEDICINE · 2024 · $652,880

## Abstract

ABSTRACT
The recent development of T cell-based cancer immunotherapies, including checkpoint blockade (anti-PD-1,
anti-CTLA-4 and others) or adoptive T cell therapy (ACT) using modified patient T cells, has led to improved
patient outcomes for a variety of cancers. However, durable responses are observed in only a fraction of patients.
Further progress can be made by studying and targeting different T cell signaling pathways, such as the T cell
receptor (TCR)-CD3 signaling pathway. T cell recognition of antigen by the TCR and the resulting proximal
signaling through surrounding CD3 subunits are key steps in the initiation of tumor-killing. Previous studies that
targeted the antigen binding site of the TCR for enhancing T cell responses to tumor antigens often lead to off-
target effects and toxicity. Instead, identification of the specific extracellular interactions between the TCR and
CD3 subunits could offer precise guidance for the development of immunotherapeutic strategies that modulate
T cell immunity by targeting signaling through the TCR-CD3 complex without altering antigen-specificity. Our
preliminary data showed that mutating residues in the constant domain of the TCR resulted in altered T cell
cytokine responses. Based on our preliminary data, our hypothesis is that by modulating TCR-CD3 signaling,
immune-mediated cytotoxicity to tumor antigens can be enhanced without losing specificity for the cancer
antigen. To test our hypothesis, in Aim 1 we will use an in vitro retroviral TCR display method, a novel CD3-
tetramer assay, and an in-silico structure-based TCR design approach to identify signal-enhancing TCR mutants
that enable T cells to mediate more effective in vitro tumor killing. In Aim 2, we will identify the mechanisms that
drive altered signaling evidenced in select TCR signal-enhancing and signal-hampering mutants. In Aim 3,
identified mutations will be introduced into gp100-specific TCRs with different antigen affinities and their in vitro
and in vivo tumor killing efficacy will be analyzed to characterize the tumor killing potential, T cell differentiation,
and T cell exhaustion patterns of new signal-enhancing T cell clones with the goal of developing a new strategy
for effective T cell therapies against cancer.

## Key facts

- **NIH application ID:** 10880853
- **Project number:** 1R01CA284604-01A1
- **Recipient organization:** NEW YORK UNIVERSITY SCHOOL OF MEDICINE
- **Principal Investigator:** MICHELLE KROGSGAARD
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $652,880
- **Award type:** 1
- **Project period:** 2024-04-10 → 2029-03-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10880853

## Citation

> US National Institutes of Health, RePORTER application 10880853, Optimizing TCR-CD3 signaling for immunotherapy of cancer (1R01CA284604-01A1). Retrieved via AI Analytics 2026-05-28 from https://api.ai-analytics.org/grant/nih/10880853. Licensed CC0.

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