# Construction of functional human CAR recognizing live pancreatic beta cells

> **NIH NIH P20** · SANFORD RESEARCH/USD · 2022 · $298,988

## Abstract

Type 1 diabetes (T1D), autoimmune pediatric disease, occurs due to hyperactivation of self-reactive T 
lymphocytes, which destroy pancreatic β cells. Suppression by regulatory T cells (Tregs) is a key mechanism 
limiting such autoreactivity. T1D patients have quantitative and functional Tregs defects. New clinical trials, 
transferring autologous ex vivo-expanded Tregs into T1D patients, showed signs of success. 
Our proposal targets major improvement of the current Treg therapies. We aim to create and validate a 
novel, antigen-specific cellular therapy for T1D. Hence, we designed β cell-specific Chimeric Antigen 
Receptors (CARs), to be optimized and tested herein. β cell-specificity is conferred by the scFv fragment of 
an antibody recognizing unique human β cell marker NTPDase3. We hypothesize that engagement of β cellspecific 
CAR(s) on T cells will induce proper CAR signaling and cause activation and expansion of CAR T 
cells. When such signaling occurs in CAR-bearing Tregs, additionally modified for sustained 
immunosuppression through augmented expression of the essential for Tregs function transcription factors 
Foxp3 and Eos, it will enable local long-lasting antigen-dependent silencing of diabetogenic autoimmunity. 
This hypothesis will be tested in the following Specific Aims: 
Aim 1. Design and produce β cell-specific CAR(s), optimize their binding strength, and interrogate 
their signaling potential in vitro. As either too strong or weak binding of our CARs to β cell antigen will 
likely provide inadequate activation, we will model our paratope in silico to design mutated variants with 
optimal antigen binding. Optimized mutant scFv-CAR constructs will be analyzed and CARs that provide 
constitutive antigen-specific signaling without subsequent T cells exhaustion will be prioritized. 
Aim 2. Develop and test strategies allowing for maintenance of continued stable and robust 
suppressive phenotype of β cell-specific CAR Tregs. We will test our inventive strategy creating 
autologous CAR-bearing induced T regs (iTregs) from the bulk of conventional CD4+ T cells. For that CAR 
cassettes are designed to provide constitutive ectopic expression of crucial for Tregs development and 
maintenance transcription factors Foxp3 and/or Eos. CAR iTregs will be tested functionally for their 
suppressive abilities against human β cells-reactive diabetogenic T cell clones. 
Completion of our project will confirm the feasibility of β cell-specific CAR Tregs, evaluate their potency, 
and pave a road for future pre-clinical and clinical trials of CAR Tregs in T1D.

## Key facts

- **NIH application ID:** 10669552
- **Project number:** 5P20GM103620-10
- **Recipient organization:** SANFORD RESEARCH/USD
- **Principal Investigator:** Alexei Yurievitch Savinov
- **Activity code:** P20 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $298,988
- **Award type:** 5
- **Project period:** 2013-09-01 → 2025-08-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10669552

## Citation

> US National Institutes of Health, RePORTER application 10669552, Construction of functional human CAR recognizing live pancreatic beta cells (5P20GM103620-10). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10669552. Licensed CC0.

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