# Development of a novel platform for label-free monitoring of CAR-T cell interactions in vivo

> **NIH NIH R21** · TUFTS UNIVERSITY MEDFORD · 2024 · $177,279

## Abstract

Chimeric antigen receptor (CAR)-T cell therapy has resulted in dramatic, durable responses, especially for
patients with B cell malignancies that are refractory to standard treatment. Numerous clinical trials are focused
on improvements of the approach and applications for a much broader range of hematologic malignancies and
solid tumors. However, this targeted therapy is also associated with high relapse rates and the development of
severe toxicities that are challenging to predict. Thus, improved tools to optimize and monitor response are
critically needed. Our ultimate goal is to develop a simple, quantitative, and non-invasive point-of-care device to
monitor CAR-T cell interactions in vivo dynamically with the needed frequency. The objective of this proposal is
to demonstrate proof-of-principle of the approach and its translational potential for human studies. Our approach
relies on in vivo flow cytometry (IVFC) assessments using endogenous and exogenous sources of optical
contrast and its optimization and use to detect CAR-T/B cell interactions in vitro and in vivo. The proposed
method relies on the confocal detection of endogenous light scattering and fluorescence (label-free or LF) signals
of flowing CAR-T/B cells that are interacting. Previous studies support the hypothesis that interacting CAR-T/B
cells will likely yield unique optical signatures. We anticipate that utilization of advanced signal processing
approaches will enable us to optimize detection of these signals in the challenging milieu of flowing blood in vitro
and in vivo. Initial measurements with fluorescent protein (FP)-expressing human CAR-T and B cells spiked in
whole blood will allow us to develop, implement, and assess enhancements in hardware and software to realize
accurate LF-IVFC assessments of interacting CAR-T and B cells. (Aim 1.1). The translational potential will be
tested on blood samples isolated from patients undergoing CAR-T cell therapy at different time points over the
first month of therapy. We expect to detect variations in the numbers of interacting CAR-T and B cells using LF-
IVFC and this expectation will be corroborated with antibody-based flow cytometry measurements (Aim 1.2).
Finally, we will perform in vivo LF and FP-based IVFC measurements using mice infused with human B
lymphoblasts and CAR-T cells at concentrations known to induce variations in the relative circulating levels of
these cells. Our goal will be to demonstrate that LF-IVFC measurements can be performed with high enough
accuracy to detect dynamic changes in the numbers of interacting CAR-T and B cells over a month. We expect
to generate a robust set of data that will motivate development and application of LF-IVFC monitoring of
interacting CAR-T/B cells for dynamic, non-invasive monitoring of patients receiving CAR-T cell therapy. Such a
device may enable acquisition of information regarding individual patient responses with a combined time
resolution and sensitivity/specificity t...

## Key facts

- **NIH application ID:** 10755335
- **Project number:** 5R21CA271679-02
- **Recipient organization:** TUFTS UNIVERSITY MEDFORD
- **Principal Investigator:** Irene Georgakoudi
- **Activity code:** R21 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $177,279
- **Award type:** 5
- **Project period:** 2023-01-01 → 2025-12-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10755335

## Citation

> US National Institutes of Health, RePORTER application 10755335, Development of a novel platform for label-free monitoring of CAR-T cell interactions in vivo (5R21CA271679-02). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10755335. Licensed CC0.

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