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Enhancing Chimeric Antigen Receptor T-Cell Generation via Microfluidic Mechanoporation and Lipid Nanoparticles
Jianhua Lim1, Daniel Oh1, Makayla Cheng1
1Department of Bioengineering, University of Pennsylvania, Philadelphia, PA, 19104, USA.
Small (Weinheim an Der Bergstrasse, Germany)
|March 19, 2025
Summary
A new LNP + Squeeze platform improves non-viral Chimeric Antigen Receptor (CAR)-T cell engineering. This cost-effective method enhances CAR-T cell therapy
Area of Science:
- Biotechnology
- Immunotherapy
- Cellular Engineering
Background:
- Chimeric antigen receptor (CAR)-T cell therapy offers a revolutionary approach to cancer treatment.
- Traditional viral transduction methods for CAR-T cell manufacturing face challenges including high costs, severe side effects, and immune reactions.
- Non-viral transfection methods are being explored as safer and more cost-effective alternatives.
Purpose of the Study:
- To introduce a novel high-throughput intracellular delivery platform, LNP + Squeeze, for non-viral CAR-T cell engineering.
- To evaluate the efficiency and viability of T cells transfected using this new platform compared to existing non-viral methods.
- To demonstrate the efficacy of CAR-T cells engineered by LNP + Squeeze against cancer cells.
Main Methods:
- Development of a high-throughput intracellular delivery platform integrating microfluidic mechanoporation with lipid nanoparticle (LNP)-based delivery (LNP + Squeeze).
- Transfection of plasmid DNA (pDNA) encoding CAR into primary human T cells using the LNP + Squeeze platform.
- Assessment of transfection efficiency and cell viability in comparison to electroporation.
- Evaluation of the cytotoxic activity of engineered CAR-T cells against melanoma cells.
Main Results:
- The LNP + Squeeze platform significantly enhanced pDNA transfection efficiency in T cells.
- Cell viability was maintained at high levels using the LNP + Squeeze method, outperforming electroporation.
- Successfully engineered CAR-T cells using primary human T cells demonstrated potent cytotoxicity against melanoma cells.
Conclusions:
- The LNP + Squeeze platform provides a highly efficient and viable non-viral method for CAR-T cell engineering.
- This approach presents a promising, cost-effective, and scalable alternative to viral transduction methods.
- The developed platform has the potential to improve the accessibility and efficacy of CAR-T cell therapies for cancer treatment.
