# Development of Tunable Microenvironment-Responsive CAR T Cells Using Synthetic Gene Circuits to Enhance Potency and Safety

> **NIH NIH F30** · NORTHWESTERN UNIVERSITY · 2024 · $53,974

## Abstract

PROJECT SUMMARY
 Ovarian cancer is the most lethal malignancy of the female reproductive system, and despite advances
in understanding of the disease, therapeutic options for treating this disease are limited. Synthetic biology affords
the ability to engineer cell-based therapies, such as CAR T cells, that are programmable and targeted, and thus
provide a pathway for overcoming barriers to the treatment of solid tumors. CAR T cells have demonstrated
success in the treatment of hematological malignancies but have experienced challenges in their application to
solid tumors due to the lack of specific tumor antigens present on solid tumor cells, as well as the presence of
immunosuppressive microenvironments surrounding solid tumors. The hypoxic microenvironment ubiquitous to
solid tumors can be used as a biomarker to restrict CAR expression within a tumor, improving the safety and
specificity of solid tumor CAR T cell therapy, but so far this approach has only provided an effective anti-tumor
response at levels of profound hypoxia. Thus, strategies are needed to provide specific, anti-tumor responses in
tumors with modest levels of hypoxia that are not addressed by current approaches. To meet this need, we have
developed a hypoxia biosensor (HBS) circuit enhanced by positive feedback motifs, enabling fast transcriptional
output with minimal background signal. My objective is to engineer microenvironment-responsive CAR T cells
(Tune-Up CARs) containing this HBS circuit to improve the safety and potency of solid tumor CAR T cell therapy
for modestly hypoxic tumors. I hypothesize that the successful induction of this HBS circuit in the
engineered T cells within the tumor microenvironment will enable CAR expression at modest levels of
hypoxia and result in the tumor-localized delivery of immunostimulatory cytokines, inducing tumor
regression. In the first Specific Aim of the proposal, I will functionally implement the HBS circuit in human primary
T cells using transposon and lentiviral vectors to integrate the constructs into the T cell genome and evaluate
their performance in vitro. The second Specific Aim will focus on evaluating the engineered Tune-Up CAR T cells
to benchmark against current state-of-the-art technology in vivo. In the third Specific Aim, I will evaluate the anti-
tumor function and specificity of Tune-Up CAR T cells in vivo in a translationally relevant ovarian cancer model.
If successful, the proposed research will provide a new therapeutic technology for treating ovarian
cancer and inform engineering of new classes of hypoxia-responsive cell-based therapies.

## Key facts

- **NIH application ID:** 10995779
- **Project number:** 1F30CA275362-01A1
- **Recipient organization:** NORTHWESTERN UNIVERSITY
- **Principal Investigator:** Yannick Rene Schreiber
- **Activity code:** F30 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $53,974
- **Award type:** 1
- **Project period:** 2024-09-01 → 2026-08-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10995779

## Citation

> US National Institutes of Health, RePORTER application 10995779, Development of Tunable Microenvironment-Responsive CAR T Cells Using Synthetic Gene Circuits to Enhance Potency and Safety (1F30CA275362-01A1). Retrieved via AI Analytics 2026-05-24 from https://api.ai-analytics.org/grant/nih/10995779. Licensed CC0.

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