1. Academic Validation
  2. Scalable intracellular delivery via microfluidic vortex shedding enhances the function of chimeric antigen receptor T-cells

Scalable intracellular delivery via microfluidic vortex shedding enhances the function of chimeric antigen receptor T-cells

  • bioRxiv. 2024 Jul 13:2024.06.25.600671. doi: 10.1101/2024.06.25.600671.
Brandon J Sytsma 1 Vincent Allain 2 3 4 Struan Bourke 1 Fairuz Faizee 1 Mohsen Fathi 5 Rebecca Berdeaux 5 Leonardo M R Ferreira 1 6 7 8 W Jared Brewer 1 Lian Li 1 Fong L Pan 1 Allison G Rothrock 2 3 William A Nyberg 2 3 Zhongmei Li 2 3 Leah H Wilson 1 Justin Eyquem 1 2 3 9 10 11 12 Ryan S Pawell 1
Affiliations

Affiliations

  • 1 Indee Labs, Berkeley, CA, USA.
  • 2 Gladstone-UCSF Institute of Genomic Immunology, San Francisco, CA, USA.
  • 3 Department of Medicine, University of California, San Francisco, San Francisco, CA, USA.
  • 4 Université Paris Cité, INSERM UMR976, Hôpital Saint-Louis, Paris, France.
  • 5 CellChorus, Houston, TX, United States.
  • 6 Department of Microbiology and Immunology, Medical University of South Carolina, Charleston, SC, USA.
  • 7 Department of Regenerative Medicine and Cell Biology, Medical University of South Carolina, Charleston, SC, USA.
  • 8 Hollings Cancer Center, Medical University of South Carolina, Charleston, SC, USA.
  • 9 UCSF Helen Diller Family Comprehensive Cancer Center, University of California, San Francisco, San Francisco, CA, USA.
  • 10 Institute for Human Genetics (IHG), University of California, San Francisco, San Francisco, CA, USA.
  • 11 Parker Institute for Cancer Immunotherapy, San Francisco, CA, USA.
  • 12 Department of Microbiology and Immunology, University of California, San Francisco, San Francisco, CA, USA.
Abstract

Adoptive chimeric antigen receptor T-cell (CAR-T) therapy is transformative and approved for hematologic malignancies. It is also being developed for the treatment of solid tumors, autoimmune disorders, heart disease, and aging. Despite unprecedented clinical outcomes, CAR-T and Other engineered cell therapies face a variety of manufacturing and safety challenges. Traditional methods, such as lentivirus transduction and electroporation, result in random integration or cause significant cellular damage, which can limit the safety and efficacy of engineered cell therapies. We present hydroporation as a gentle and effective alternative for intracellular delivery. Hydroporation resulted in 1.7- to 2-fold higher CAR-T yields compared to electroporation with superior cell viability and recovery. Hydroporated cells exhibited rapid proliferation, robust target Cell Lysis, and increased pro-inflammatory and regulatory cytokine secretion in addition to improved CAR-T yield by day 5 post-transfection. We demonstrate that scaled-up hydroporation can process 5 x 108 cells in less than 10 s, showcasing the platform as a viable solution for high-yield CAR-T manufacturing with the potential for improved therapeutic outcomes.

Keywords

AAV; CAR-T; CRISPR; Cell Therapy; Cytokine; Gene editing; Intracellular delivery; Microfluidics; Serial killing; Vortex shedding.

Figures
Products
  • Cat. No.
    Product Name
    Description
    Target
    Research Area
  • HY-101570
    99.70%, DNA-PK Inhibitor