# Hydrogel microparticle technology for high-throughout screening of chimeric antigen receptor-T cells based on single cell effector function

> **NIH NIH R44** · PARTILLION BIOSCIENCE CORPORATION · 2023 · $754,792

## Abstract

ABSTRACT
Engineered cell therapies are becoming a pillar of medicine, along with molecular and genetic interventions. In
particular, chimeric antigen receptor (CAR)-T cell therapies have had a number of FDA approvals in the last 5
years targeting hematologic malignancies. To extend the success of these therapies, especially for the treatment
of solid tumors, a more thorough understanding of how CAR structure is linked to CAR-T cell function is
necessary. Notably, widespread tools to analyze and select single CAR-T cells from a large population based
on functional properties, such as secreted products or cytolytic activity, are critically lacking. Single-cell functional
assays can enable screening a library of CAR designs introduced into a pool of cells to identify rare functional
cells and sort these cells to recover CAR designs associated with important effector functions. If conducted in
high throughput, thousands of constructs can be screened with hundreds of individual events per construct to
have robust statistical accuracy in the linkage between function and sequence. In addition, if single-cell functional
information can be tied to transcriptomic information, pathway analysis associated with strong effector functions
can be performed, identifying other gene targets that improve function, even in the presence of
immunosuppressive, exhaustion-prone, or other microenvironments associated with solid tumors. Partillion’s
nanovial technology provides a new approach to measure the function of single cells using widely available
fluorescence activated cell sorters (FACS) and single-cell sequencing instruments, which we aim to apply to cell
therapy discovery. Here, we propose to develop single-cell secretion and cell-killing assays compatible with the
nanovials to introduce a new product that would enable scaled cell therapy discovery workflows from millions of
cells. We aim to identify optimal nanovial formulations and procedures to measure both cytokine production and
cytolytic functions from the same cells, and then link single-cell transcriptomic information with this functional
readout. We also will engineer nanovials to better recapitulate the tumor microenvironment, acting as an artificial
antigen-presenting target cell with combinations of antigen and immunosuppressive signals. The approach
should be applicable broadly beyond CAR-T cell therapies to other chimeric receptors in natural killer cells, and
macrophages, or in finding engineered T cell receptors. Ultimately, more access to sophisticated cell selection
approaches can lead to therapies that are both lower in price and more effective as well as expanding the scope
of applications to un-explored therapeutic areas.

## Key facts

- **NIH application ID:** 10604170
- **Project number:** 1R44GM149102-01
- **Recipient organization:** PARTILLION BIOSCIENCE CORPORATION
- **Principal Investigator:** Joseph de Rutte
- **Activity code:** R44 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2023
- **Award amount:** $754,792
- **Award type:** 1
- **Project period:** 2023-09-06 → 2025-07-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10604170

## Citation

> US National Institutes of Health, RePORTER application 10604170, Hydrogel microparticle technology for high-throughout screening of chimeric antigen receptor-T cells based on single cell effector function (1R44GM149102-01). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10604170. Licensed CC0.

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