Related Experiment Video
Updated: Jul 21, 2025
Manufacturing Chimeric Antigen Receptor CAR T Cells for Adoptive Immunotherapy
Published on: December 17, 2019
PASCAR: a multiscale framework to explore the design space of constitutive and inducible CAR T cells
Harshana Rajakaruna1, Milie Desai2, Jayajit Das3,4,5
1The Steve and Cindy Rasmussen Institute for Genomics, The Abigail Wexner Research Institute, Nationwide Children's Hospital, Columbus, OH, USA.
Abstract:
CAR T cells are engineered to bind and destroy tumor cells by targeting overexpressed surface antigens. However, healthy cells expressing lower abundances of these antigens can also be lysed by CAR T cells. Various CAR T cell designs increase tumor cell elimination, whereas reducing damage to healthy cells. However, these efforts are costly and labor-intensive, constraining systematic exploration of potential hypotheses. We develop a protein abundance structured population dynamic model for CAR T cells (PASCAR), a framework that combines multiscale population dynamic models and multi-objective optimization approaches with data from cytometry and cytotoxicity assays to systematically explore the design space of constitutive and tunable CAR T cells. PASCAR can quantitatively describe in vitro and in vivo results for constitutive and inducible CAR T cells and can successfully predict experiments outside the training data. Our exploration of the CAR design space reveals that optimal CAR affinities in the intermediate range of dissociation constants effectively reduce healthy cell lysis, whereas maintaining high tumor cell-killing rates. Furthermore, our modeling offers guidance for optimizing CAR expressions in synthetic notch CAR T cells. PASCAR can be extended to other CAR immune cells.
Insights
We developed PASCAR, a model to optimize CAR T cell therapy by balancing tumor cell killing and reducing healthy cell damage. This computational framework enhances CAR T cell design for improved safety and efficacy.
Area of Science:
- Immunotherapy
- Computational Biology
- Systems Biology
Background:
- Chimeric Antigen Receptor (CAR) T cells offer potent anti-tumor immunity by targeting tumor-specific antigens.
- CAR T cell therapy faces challenges with on-target, off-tumor toxicity due to antigen expression on healthy tissues.
- Current CAR T cell design optimization is limited by cost and labor, hindering systematic exploration.
Purpose of the Study:
- To develop a computational framework, PASCAR, for systematic exploration of CAR T cell design space.
- To optimize CAR T cell therapies for enhanced tumor cell elimination while minimizing healthy cell damage.
- To provide guidance for designing both constitutive and tunable CAR T cells.
Main Methods:
- Developed a protein abundance structured population dynamic model for CAR T cells (PASCAR).
- Integrated multiscale population dynamics with multi-objective optimization using cytometry and cytotoxicity data.
- Validated PASCAR's predictive capability using in vitro and in vivo experimental data.
Main Results:
- PASCAR quantitatively describes CAR T cell behavior across different designs and experimental settings.
- Identified optimal CAR affinities within an intermediate range of dissociation constants to reduce healthy cell lysis.
- Demonstrated PASCAR's ability to predict experimental outcomes beyond the training data, including synthetic notch CAR T cells.
Conclusions:
- PASCAR provides a powerful computational tool for systematic exploration and optimization of CAR T cell designs.
- Intermediate CAR affinities are key to balancing anti-tumor efficacy with reduced off-tumor toxicity.
- The PASCAR framework can be extended to other CAR-based immunotherapies.
