Related Experiment Video
Updated: Sep 17, 2025
Droplet-based Cytotoxicity Assay to Assess Chimeric Antigen Receptor T cells at the Single-cell Level
Published on: March 14, 2025
Bioengineered immunocompetent preclinical trial-on-chip tool enables screening of CAR T cell therapy for leukaemia
Chao Ma1,2,3, Huishu Wang1, Lunan Liu1
1Department of Mechanical and Aerospace Engineering, Tandon School of Engineering, New York University, Brooklyn, NY, USA.
Abstract:
Chimeric antigen receptor (CAR) T cell immunotherapy is promising for treatment of blood cancers; however, clinical benefits remain unpredictable, necessitating development of optimal CAR T cell products. Unfortunately, current preclinical evaluation platforms are inadequate owing to their limited physiological relevance to humans. Here we engineer an organotypic immunocompetent chip that recapitulates microarchitectural and pathophysiological characteristics of human leukaemia bone marrow stromal and immune niches for CAR T cell therapy modelling. This leukaemia chip empowers real-time spatiotemporal monitoring of CAR T cell functionality, including T cell extravasation, recognition of leukaemia, immune activation, cytotoxicity and killing. We use our chip to model clinically observed heterogeneous responses such as remission, resistance and relapse under CAR T cell therapy and map factors that drive therapeutic success or failure. Finally, we demarcate functional performance of CAR T cells produced from different healthy donors and patients with cancer, with various CAR designs and protocols, systematically and multidimensionally. Together, our chip introduces an enabling '(pre-)clinical-trial-on-chip' tool for CAR T cell development, which may translate to personalized therapies and improved clinical decision-making.
Insights
A new "pre-clinical-trial-on-chip" platform models human leukemia bone marrow niches to optimize chimeric antigen receptor (CAR) T cell therapy, predicting treatment responses and guiding personalized medicine.
Area of Science:
- Biomedical Engineering
- Immunotherapy Research
- Cancer Modeling
Background:
- Chimeric antigen receptor (CAR) T cell immunotherapy shows promise for blood cancers but faces unpredictable clinical benefits.
- Current preclinical models lack the physiological relevance needed to optimize CAR T cell products.
Purpose of the Study:
- To engineer an organotypic immunocompetent chip that mimics human leukemia bone marrow microenvironments.
- To establish a platform for real-time, spatiotemporal monitoring of CAR T cell functionality in a human-relevant context.
Main Methods:
- Development of an organotypic immunocompetent chip replicating human leukemia bone marrow stromal and immune niches.
- Utilizing the chip for real-time monitoring of CAR T cell extravasation, leukemia recognition, immune activation, and cytotoxicity.
- Modeling heterogeneous clinical responses including remission, resistance, and relapse.
Main Results:
- The chip successfully models clinically observed heterogeneous responses to CAR T cell therapy.
- Factors driving therapeutic success or failure in CAR T cell treatment were mapped.
- Functional performance of CAR T cells from diverse donors, cancer patients, and varying designs/protocols was systematically evaluated.
Conclusions:
- The developed leukemia chip serves as an effective '(pre-)clinical-trial-on-chip' tool for CAR T cell development.
- This platform has the potential to advance personalized CAR T cell therapies and improve clinical decision-making.
