Integrated strategy for developing next-generation CAR-T therapies using CRISPR and organoid platforms.
Technology platform
Core input
Process & methodology
Key output
Contribution to CAR-T therapy development
CRISPR screening
Patient-derived gastric cancer organoids, genome-wide sgRNA library.
Genome-wide CRISPR knockout screening in organoids, followed by NGS analysis of enriched/depleted sgRNAs.
1. Validated novel therapeutic targets (e.g., membrane protein X).2. Potential targets for overcoming resistance.
Provides a target blueprint: Enables systematic, unbiased discovery of ideal new targets for CAR design, providing source innovation.
Organoid modeling & co-culture
1. Novel targets from screening.2. Candidate CAR-T cells.
1. Generation of organoids expressing the novel target.2. In vitro co-culture with candidate CAR-T cells.3. Real-time imaging, cytotoxicity assays, cytokine profiling.
1. Quantitative cytotoxicity data of CAR-T cells.2. Assessment of tumor specificity (on-target activity).3. Early warning of “on-target, off-tumor” toxicity risk.
Provides a predictive preclinical platform: High-throughput validation of candidate CAR-T efficacy and safety in a system that closely mimics the human TME, significantly de-risking clinical translation.
CRISPR engineering of CAR-T
1. Validated functional targets.2. Patient T cells.
1. Vector delivery: Introduction of CAR genes targeting novel antigens.2. Gene editing: Knockout of immune checkpoints (e.g., PD-1, A2AR) to enhance function.
Provides an optimized “living drug”: Translates upstream discoveries into potent weapons against solid tumors. Generates more potent and persistent CAR-T products via multiplexed gene editing.
MZ: Writing—original draft. QS: Visualization. QZ: Writing—review & editing. XY: Conceptualization, Writing—original draft, Writing—review & editing, Supervision. All authors have read and agreed to the published version of the manuscript.
Open Exploration maintains a neutral stance on jurisdictional claims in published institutional affiliations and maps. All opinions expressed in this article are the personal views of the author(s) and do not represent the stance of the editorial team or the publisher.
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