دورية أكاديمية
A versatile CRISPR-Cas13d platform for multiplexed transcriptomic regulation and metabolic engineering in primary human T cells.
العنوان: | A versatile CRISPR-Cas13d platform for multiplexed transcriptomic regulation and metabolic engineering in primary human T cells. |
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المؤلفون: | Tieu V; Department of Bioengineering, Stanford University, Stanford, CA 94305, USA; Center for Cancer Cell Therapy, Stanford Cancer Institute, Stanford University School of Medicine, Stanford, CA 94305, USA., Sotillo E; Center for Cancer Cell Therapy, Stanford Cancer Institute, Stanford University School of Medicine, Stanford, CA 94305, USA., Bjelajac JR; Center for Cancer Cell Therapy, Stanford Cancer Institute, Stanford University School of Medicine, Stanford, CA 94305, USA., Chen C; Department of Chemical Engineering, Stanford University, Stanford, CA 94305, USA., Malipatlolla M; Center for Cancer Cell Therapy, Stanford Cancer Institute, Stanford University School of Medicine, Stanford, CA 94305, USA., Guerrero JA; Center for Cancer Cell Therapy, Stanford Cancer Institute, Stanford University School of Medicine, Stanford, CA 94305, USA., Xu P; Center for Cancer Cell Therapy, Stanford Cancer Institute, Stanford University School of Medicine, Stanford, CA 94305, USA., Quinn PJ; Center for Cancer Cell Therapy, Stanford Cancer Institute, Stanford University School of Medicine, Stanford, CA 94305, USA., Fisher C; Center for Cancer Cell Therapy, Stanford Cancer Institute, Stanford University School of Medicine, Stanford, CA 94305, USA., Klysz D; Center for Cancer Cell Therapy, Stanford Cancer Institute, Stanford University School of Medicine, Stanford, CA 94305, USA., Mackall CL; Center for Cancer Cell Therapy, Stanford Cancer Institute, Stanford University School of Medicine, Stanford, CA 94305, USA; Department of Pediatrics, Stanford University School of Medicine, Stanford, CA 94305, USA; Department of Medicine, Stanford University School of Medicine, Stanford, CA 94305, USA. Electronic address: cmackall@stanford.edu., Qi LS; Department of Bioengineering, Stanford University, Stanford, CA 94305, USA; Sarafan ChEM-H, Stanford University, Stanford, CA 94305, USA; Chan Zuckerberg Biohub, San Francisco, San Francisco, CA 94080, USA. Electronic address: stanley.qi@stanford.edu. |
المصدر: | Cell [Cell] 2024 Feb 29; Vol. 187 (5), pp. 1278-1295.e20. Date of Electronic Publication: 2024 Feb 21. |
نوع المنشور: | Journal Article |
اللغة: | English |
بيانات الدورية: | Publisher: Cell Press Country of Publication: United States NLM ID: 0413066 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1097-4172 (Electronic) Linking ISSN: 00928674 NLM ISO Abbreviation: Cell Subsets: MEDLINE |
أسماء مطبوعة: | Publication: Cambridge, Ma : Cell Press Original Publication: Cambridge, MIT Press. |
مواضيع طبية MeSH: | Metabolic Engineering*/methods , T-Lymphocytes*, Humans ; Gene Expression Profiling ; RNA ; Transcriptome |
مستخلص: | CRISPR technologies have begun to revolutionize T cell therapies; however, conventional CRISPR-Cas9 genome-editing tools are limited in their safety, efficacy, and scope. To address these challenges, we developed multiplexed effector guide arrays (MEGA), a platform for programmable and scalable regulation of the T cell transcriptome using the RNA-guided, RNA-targeting activity of CRISPR-Cas13d. MEGA enables quantitative, reversible, and massively multiplexed gene knockdown in primary human T cells without targeting or cutting genomic DNA. Applying MEGA to a model of CAR T cell exhaustion, we robustly suppressed inhibitory receptor upregulation and uncovered paired regulators of T cell function through combinatorial CRISPR screening. We additionally implemented druggable regulation of MEGA to control CAR activation in a receptor-independent manner. Lastly, MEGA enabled multiplexed disruption of immunoregulatory metabolic pathways to enhance CAR T cell fitness and anti-tumor activity in vitro and in vivo. MEGA offers a versatile synthetic toolkit for applications in cancer immunotherapy and beyond. Competing Interests: Declaration of interests The authors have filed a patent related to this work. E.S. consults for Lyell Immunopharma, Lepton Pharmaceuticals, and Galaria. C.L.M. is a cofounder of Lyell Immunopharma, CARGO Therapeutics, and Link Cell Therapies and consults for Lyell, CARGO, Link, Apricity, Nektar, Immatics, Ensoma, Mammoth, Glaxo Smith Kline, Bristol Myers Squibb, and RedTree Capital. L.S.Q. is a founder of Epic Bio and scientific advisor of Laboratory of Genomic Research and Kytopen Corp. (Copyright © 2024 Elsevier Inc. All rights reserved.) |
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معلومات مُعتمدة: | P30 CA124435 United States CA NCI NIH HHS; R21 CA270609 United States CA NCI NIH HHS |
فهرسة مساهمة: | Keywords: CAR T cells; CRISPR; Cas13; RNA targeting; T cell exhaustion; cancer immunotherapy; cell therapy; gene editing; metabolic engineering; synthetic biology |
المشرفين على المادة: | 63231-63-0 (RNA) |
تواريخ الأحداث: | Date Created: 20240222 Date Completed: 20240304 Latest Revision: 20240328 |
رمز التحديث: | 20240329 |
مُعرف محوري في PubMed: | PMC10965243 |
DOI: | 10.1016/j.cell.2024.01.035 |
PMID: | 38387457 |
قاعدة البيانات: | MEDLINE |
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