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Optimizing Ex Vivo CAR-T Cell-mediated Cytotoxicity Assay Through Multimodality Imaging

Poster
CAR-T cells, CAR-Target, CD19, CD20, HER2

Society for Immunotherapy of Cancer (SITC) 2023 Annual Meeting

San Diego, California, United States

November 03, 2023

Abstract

CAR-T cell–based therapies have demonstrated remarkable efficacy in treating malignant cancers, especially liquid tumors, and are increasingly being evaluated in clinical trials for solid tumors. With FDA’s initiative for advancing alternative methods for drug discovery and development, full human ex vivo assays are increasingly essential for precision CAR-T development. However, prevailing ex vivo CAR-T cell–mediated cytotoxicity assays are limited by their use of radioactive materials, lack of real-time measurement, low throughput, and automatability, among others. To address these limitations, we optimized the assay using multimodality imaging methods, including bioluminescence, impedance tracking, phase contrast, and fluorescence, to track CAR-T cells cocultured with CD19, CD20, and HER2 luciferase reporter cancer cells in real-time. Additionally, we varied the ratio of CAR-T cells to cancer cells to determine optimal cytotoxicity readouts. Our findings demonstrated that the CAR-T cell group effectively attacked cancer cells, and the optimized assay provided superior temporal and spatial precision measurements of ex vivo CAR-T killing of cancer cells, confirming the reliability, consistency, and high throughput of the optimized assay.

Download the poster to explore our data on the optimization of a CAR-T cell-mediated cytotoxicity assay

Download

Watch the poster presentation

Presenter

John Foulke, headshot

John Foulke, MS

Lead Biologist, ATCC

John Foulke is a Lead Biologist in the Immuno-Oncology group in the R&D department at ATCC. John joined the ATCC cell biology R&D group in 2008, and he has led many projects centered on the development of novel cell lines and cell-based reporter systems to support cancer research community. His work is mainly focused on developing innovative cell models for research and drug discovery in the immuno-oncology field.