Related Experiment Video
Updated: Sep 19, 2025
Non-Viral Engineering of Primary Human T Cells via Homology-Mediated End-Joining Targeted Integration of Large DNA Templates
Published on: May 9, 2025
Syngeneic CAR T cells engineered for persistent delivery of desired proteins
Sherri L Newmyer1, Harikrishnan Radhakrishnan1, Harold S Javitz2
1Biosciences Division, SRI International, Menlo Park, CA 94025, United States of America.
Abstract:
Mouse primary T cells have been engineered as a platform using chimeric antigen receptors (CARs) to induce the synthesis of desired proteins at the disease site. This approach allows for the use of immunocompetent syngeneic tumor models to evaluate the CAR T cells' function within the context of a fully functioning immune system. Current efforts to evaluate cell-based technologies typically rely on xenograft tumor models in immunodeficient mice, which provide early feasibility data but may not fully capture the immune effects present in the tumor microenvironment. In this study, a primary T-cell-based system for site-specific protein expression has been translated from human T cells to mouse T cells, allowing for the use of an immunocompetent syngeneic tumor model. A lentivector transduction, effective in human T cells, was adapted to engineer mouse T cells. CD4 and CD8 CAR T cell subsets were engineered separately and evaluated in immunocompetent mice for site-specific expression of the desired proteins. Co-expression of membrane-bound interleukin 15 (mbIL15) on the T cells enhanced intratumoral accumulation of both CD4 and CD8 CAR T cells and supported their delivery function. Validation of this platform in syngeneic models will enable efficacy assessments beyond solid tumors and allow for the evaluation of immune-related toxicities arising from interactions between the therapeutic protein, CAR T cells, and the host immune system.
Insights
Researchers engineered mouse T cells with chimeric antigen receptors (CARs) for site-specific protein synthesis. This CAR T-cell platform enables evaluation in immunocompetent models, advancing cancer therapy research.
Area of Science:
- Immunology
- Cell Therapy
- Biotechnology
Background:
- Current cell-based therapy evaluations often use immunodeficient mice with xenografts, limiting immune microenvironment insights.
- A need exists for advanced platforms to assess therapeutic efficacy in a fully functioning immune system.
Purpose of the Study:
- To engineer primary mouse T cells with chimeric antigen receptors (CARs) for site-specific protein expression.
- To adapt a human T-cell CAR platform for use in immunocompetent syngeneic mouse models.
- To evaluate the efficacy of engineered CAR T cells in a relevant tumor microenvironment.
Main Methods:
- Primary mouse T cells were engineered using lentiviral transduction to express CARs.
- CD4 and CD8 CAR T cell subsets were generated and assessed in vivo.
- Co-expression of membrane-bound interleukin 15 (mbIL15) was investigated for enhancing T cell function.
Main Results:
- The CAR T-cell platform was successfully translated from human to mouse T cells.
- Engineered mouse CAR T cells demonstrated site-specific protein expression in syngeneic models.
- Co-expression of mbIL15 improved intratumoral accumulation and delivery function of CAR T cells.
Conclusions:
- This engineered mouse CAR T-cell platform provides a robust system for evaluating cell-based therapies in immunocompetent hosts.
- The platform facilitates assessment of therapeutic protein efficacy and immune-related toxicities.
- This approach advances the preclinical evaluation of CAR T-cell therapies beyond traditional xenograft models.
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