Related Experiment Video
Updated: Jul 21, 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
Genetically Stable and Scalable Nanoengineering of Human Primary T Cells via Cell Mechanoporation
Jeongsoo Hur1, Hyelee Kim1,2, Uijin Kim3
1Department of Bioengineering, Korea University, Seoul 02841, Republic of Korea.
Abstract:
Effective tumor regression has been observed with chimeric antigen receptor (CAR) T cells; however, the development of an affordable, safe, and effective CAR-T cell treatment remains a challenge. One of the major obstacles is that the suboptimal genetic modification of T cells reduces their yield and antitumor activity, necessitating the development of a next-generation T cell engineering approach. In this study, we developed a nonviral T cell nanoengineering system that allows highly efficient delivery of diverse functional nanomaterials into primary human T cells in a genetically stable and scalable manner. Our platform leverages the unique cell deformation and restoration process induced by the intrinsic inertial flow in a microchannel to create nanopores in the cellular membrane for macromolecule internalization, leading to effective transfection with high scalability and viability. The proposed approach demonstrates considerable potential as a practical alternative technique for improving the current CAR-T cell manufacturing process.
Insights
This study introduces a novel nonviral nanoengineering system for T cell modification, enhancing CAR-T cell therapy production. This scalable method improves T cell yield and antitumor activity for more effective cancer treatments.
Area of Science:
- Biotechnology
- Cellular Engineering
- Immunotherapy
Background:
- Chimeric antigen receptor (CAR) T cell therapy shows promise for tumor regression but faces challenges in cost, safety, and efficacy.
- Suboptimal genetic modification of T cells hinders their yield and antitumor capabilities, necessitating advanced engineering strategies.
Purpose of the Study:
- To develop a novel, nonviral T cell nanoengineering platform for efficient and scalable genetic modification of primary human T cells.
- To improve the manufacturing process of CAR-T cell therapy by enhancing T cell yield and functionality.
Main Methods:
- A microfluidic system utilizing inertial flow to induce transient nanopores in the T cell membrane for macromolecule delivery.
- Nonviral delivery of functional nanomaterials into primary human T cells, ensuring genetic stability and high viability.
Main Results:
- Highly efficient delivery of diverse functional nanomaterials into primary human T cells.
- Demonstrated scalability and high cell viability of the nanoengineering approach.
- Potential for improved CAR-T cell manufacturing with enhanced yield and antitumor activity.
Conclusions:
- The developed nonviral T cell nanoengineering system offers a scalable and efficient method for T cell modification.
- This platform presents a promising alternative for advancing CAR-T cell therapy manufacturing.
- The approach addresses key challenges in current T cell engineering, paving the way for improved cancer immunotherapies.
