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

Autor:	Sytsma BJ; Indee Labs, Berkeley, CA, USA., Allain V; Gladstone-UCSF Institute of Genomic Immunology, San Francisco, CA, USA.; Department of Medicine, University of California, San Francisco, San Francisco, CA, USA.; Université Paris Cité, INSERM UMR976, Hôpital Saint-Louis, Paris, France., Bourke S; Indee Labs, Berkeley, CA, USA., Faizee F; Indee Labs, Berkeley, CA, USA., Fathi M; CellChorus, Houston, TX, United States., Berdeaux R; CellChorus, Houston, TX, United States., Ferreira LMR; Indee Labs, Berkeley, CA, USA.; Department of Microbiology and Immunology, Medical University of South Carolina, Charleston, SC, USA.; Department of Regenerative Medicine and Cell Biology, Medical University of South Carolina, Charleston, SC, USA.; Hollings Cancer Center, Medical University of South Carolina, Charleston, SC, USA., Brewer WJ; Indee Labs, Berkeley, CA, USA., Li L; Indee Labs, Berkeley, CA, USA., Pan FL; Indee Labs, Berkeley, CA, USA., Rothrock AG; Gladstone-UCSF Institute of Genomic Immunology, San Francisco, CA, USA.; Department of Medicine, University of California, San Francisco, San Francisco, CA, USA., Nyberg WA; Gladstone-UCSF Institute of Genomic Immunology, San Francisco, CA, USA.; Department of Medicine, University of California, San Francisco, San Francisco, CA, USA., Li Z; Gladstone-UCSF Institute of Genomic Immunology, San Francisco, CA, USA.; Department of Medicine, University of California, San Francisco, San Francisco, CA, USA., Wilson LH; Indee Labs, Berkeley, CA, USA., Eyquem J; Indee Labs, Berkeley, CA, USA.; Gladstone-UCSF Institute of Genomic Immunology, San Francisco, CA, USA.; Department of Medicine, University of California, San Francisco, San Francisco, CA, USA.; UCSF Helen Diller Family Comprehensive Cancer Center, University of California, San Francisco, San Francisco, CA, USA.; Institute for Human Genetics (IHG), University of California, San Francisco, San Francisco, CA, USA.; Parker Institute for Cancer Immunotherapy, San Francisco, CA, USA.; Department of Microbiology and Immunology, University of California, San Francisco, San Francisco, CA, USA., Pawell RS; Indee Labs, Berkeley, CA, USA.
Jazyk:	angličtina
Zdroj:	BioRxiv : the preprint server for biology [bioRxiv] 2024 Jul 13. Date of Electronic Publication: 2024 Jul 13.
DOI:	10.1101/2024.06.25.600671
Abstrakt:	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 10 8 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. Competing Interests: B.J.S., S.B., F.F., L.H.W., and R.S.P. are or were employed by and have an equity interest in Indee Labs. L.M.R.F., J.B., L.L., F.L.P. and J.E. are or were consultants to Indee Labs. R.S.P. is an investor in and a venture partner at both Pioneer Fund and Axial, which have a financial interest in Indee Labs. The Ferreira Lab received support from Indee Labs as a subaward from the National Institute of Diabetes and Digestive and Kidney Diseases (Grant No. 1R43DK133029-01). Indee Labs has a commercial interest in developing patents related to Hydropore™ (WO2016109864A1 & WO2019084624A1). M.F. and R.B. are or were employed by and have an equity interest in CellChorus. CellChorus received support from the National Center for Translational Sciences (R44TR005137) and the National Institute of General Medical Sciences (R44GM149106) of the National Institutes of Health and the National Science Foundation (NSF2229323). CellChorus has a commercial interest in developing the TIMING assay. J.E. is a compensated co-founder at Mnemo Therapeutics; owns stocks in Mnemo Therapeutics and Cytovia Therapeutics; is a compensated scientific advisor for Enterome, Treefrog Therapeutics and Resolution Therapeutics; and is a holder of patents pertaining to but not resulting from this work. The Eyquem Lab received research support from Cytovia Therapeutic, Mnemo Therapeutics, Takeda and Indee Labs as a subaward from the National Cancer Institute (Contract No. 7591022C00053).
Databáze:	MEDLINE
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