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Engineering Efficient CAR-T Cells via Electroactive Nanoinjection.

Ali-Reza Shokouhi1,2, Yaping Chen1,2, Hao Zhe Yoh1,2

  • 1Monash Institute of Pharmaceutical Sciences, Monash University, 381 Royal Parade, Parkville, VIC, 3052, Australia.

Advanced Materials (Deerfield Beach, Fla.)
|July 12, 2023
PubMed
Summary
This summary is machine-generated.

A novel electroactive nanoinjection (ENI) platform efficiently engineers chimeric antigen receptor (CAR)-T cells for cancer therapy. This non-viral method improves CAR gene delivery and T cell viability compared to traditional techniques.

Keywords:
CAR-T cellscancer immunotherapynano-electroporationnanoneedles and nanotubesnon-viral transfection

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Area of Science:

  • Cellular Engineering
  • Immunotherapy
  • Nanotechnology

Background:

  • Chimeric antigen receptor (CAR)-T cell therapy shows promise for blood disorders and cancers.
  • Conventional genetic engineering of T cells faces challenges like low viability and high costs.
  • Viral methods and bulk electroporation (BEP) have limitations in efficiency and safety.

Purpose of the Study:

  • To develop a novel non-viral platform for efficient CAR-T cell engineering.
  • To overcome the limitations of existing gene delivery methods for primary human T cells.
  • To assess the efficiency, viability, and functionality of CAR-T cells engineered by the new platform.

Main Methods:

  • Development of an electroactive nanoinjection (ENI) platform utilizing vertically configured electroactive nanotubes.
  • Delivery of CAR genes into primary human T cells using the ENI platform.
  • Comparison of ENI with bulk electroporation (BEP) in terms of transfection efficiency and cell viability.
  • Assessment of CAR-T cell cytotoxicity against target lymphoma cells.

Main Results:

  • The ENI platform achieved efficient CAR gene delivery (68.7%) and expression (43.3%) in T cells with high cell viability (>90%).
  • ENI demonstrated a threefold higher CAR transfection efficiency compared to BEP (43.3% vs. 16.3% GFP expression).
  • ENI-engineered CAR-T cells exhibited significant cytotoxicity (86.9%) against target lymphoma cells.

Conclusions:

  • The ENI platform offers an efficient and minimally perturbing method for engineering CAR-T cells.
  • This non-viral approach enhances CAR gene delivery and maintains T cell functionality.
  • The ENI platform holds significant potential for ex vivo cell engineering in CAR-T cell therapy and other immunotherapies.