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. |
|---|---|
| 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.) |
| References: | Nat Immunol. 2020 Aug;21(8):927-937. (PMID: 32632289) Nature. 2021 Sep;597(7878):720-725. (PMID: 34489594) Nat Biotechnol. 2023 Jul 3;:. (PMID: 37400521) Mol Ther Methods Clin Dev. 2018 Dec 31;12:145-156. (PMID: 30666307) J Immunother Cancer. 2018 Jul 13;6(1):71. (PMID: 30005714) Nat Biotechnol. 2023 Jan;41(1):108-119. (PMID: 35953673) Proc Natl Acad Sci U S A. 2021 Jul 27;118(30):. (PMID: 34285077) Nat Med. 2015 Jun;21(6):581-90. (PMID: 25939063) Nat Commun. 2019 Nov 19;10(1):5222. (PMID: 31745080) Cell Syst. 2023 Dec 20;14(12):1087-1102.e13. (PMID: 38091991) Nature. 2019 Mar;567(7749):530-534. (PMID: 30814732) Nat Commun. 2023 Sep 8;14(1):5545. (PMID: 37684268) Proc Natl Acad Sci U S A. 2019 Jun 18;116(25):12410-12415. (PMID: 31152140) Nat Commun. 2019 Mar 08;10(1):1136. (PMID: 30850590) Nat Commun. 2020 Mar 09;11(1):1281. (PMID: 32152313) Nature. 2016 May 19;533(7603):420-4. (PMID: 27096365) Commun Biol. 2023 Mar 28;6(1):334. (PMID: 36977923) Cell. 2021 Dec 9;184(25):6081-6100.e26. (PMID: 34861191) Nat Methods. 2017 Jul;14(7):687-690. (PMID: 28581496) Nat Immunol. 2021 Jul;22(7):809-819. (PMID: 34140679) Science. 2022 Nov 11;378(6620):eabn5647. (PMID: 36356142) Annu Rev Biochem. 2016 Jun 02;85:227-64. (PMID: 27145843) Nat Biotechnol. 2016 Feb;34(2):184-191. (PMID: 26780180) Cell. 2021 Mar 4;184(5):1262-1280.e22. (PMID: 33636129) Oncotarget. 2017 Mar 7;8(10):17002-17011. (PMID: 28199983) Cell. 2017 Sep 7;170(6):1120-1133.e17. (PMID: 28803728) Genome Biol. 2014;15(12):550. (PMID: 25516281) Nat Biotechnol. 2016 May;34(5):525-7. (PMID: 27043002) Cell. 2018 Dec 13;175(7):1958-1971.e15. (PMID: 30449619) Cell. 2020 May 14;181(4):865-876.e12. (PMID: 32353252) Nat Rev Cancer. 2021 Mar;21(3):145-161. (PMID: 33483715) J Clin Invest. 2022 Jan 4;132(1):. (PMID: 34981777) Nat Med. 2022 Apr;28(4):678-689. (PMID: 35440724) Mol Cell. 2018 Apr 19;70(2):327-339.e5. (PMID: 29551514) Nat Methods. 2023 Jan;20(1):86-94. (PMID: 36550277) Nature. 2019 Dec;576(7787):471-476. (PMID: 31827283) Cell. 2019 Aug 22;178(5):1189-1204.e23. (PMID: 31442407) Cell Rep. 2022 Aug 16;40(7):111226. (PMID: 35977479) Nat Biotechnol. 2018 Sep;36(8):765-771. (PMID: 30010673) Nature. 2017 Mar 2;543(7643):113-117. (PMID: 28225754) Proc Natl Acad Sci U S A. 2023 Mar 21;120(12):e2218632120. (PMID: 36920923) Sci Transl Med. 2022 Nov 9;14(670):eabn7336. (PMID: 36350986) Cell. 2018 Aug 9;174(4):953-967.e22. (PMID: 30033366) Nat Rev Immunol. 2019 May;19(5):324-335. (PMID: 30820043) Nat Protoc. 2021 Aug;16(8):3775-3801. (PMID: 34172973) Genome Biol. 2014;15(12):554. (PMID: 25476604) Nat Protoc. 2019 Mar;14(3):756-780. (PMID: 30710114) Immunity. 2017 Dec 19;47(6):1129-1141.e5. (PMID: 29246443) Nat Biotechnol. 2005 Nov;23(11):1399-405. (PMID: 16273073) Cell. 2006 Sep 8;126(5):995-1004. (PMID: 16959577) Cancer Cell. 2022 Nov 14;40(11):1294-1305.e4. (PMID: 36084652) Nature. 2018 Jun;558(7709):307-312. (PMID: 29849141) Trends Immunol. 2015 Apr;36(4):265-76. (PMID: 25797516) Nat Med. 2019 Sep;25(9):1341-1355. (PMID: 31501612) Cell Syst. 2015 Dec 23;1(6):417-425. (PMID: 26771021) Nat Rev Cancer. 2017 Dec;17(12):709-724. (PMID: 29059149) Nat Rev Clin Oncol. 2020 Oct;17(10):611-629. (PMID: 32514148) Nature. 2003 Jan 23;421(6921):388-92. (PMID: 12540904) Proc Natl Acad Sci U S A. 2021 Apr 20;118(16):. (PMID: 33859042) Nat Methods. 2012 Mar 04;9(4):357-9. (PMID: 22388286) Mucosal Immunol. 2019 Sep;12(5):1104-1117. (PMID: 31285535) BMC Res Notes. 2011 Oct 20;4:427. (PMID: 22011438) Nat Med. 2018 May;24(5):563-571. (PMID: 29713085) Nat Biotechnol. 2022 Dec;40(12):1807-1813. (PMID: 35773341) Nat Rev Immunol. 2015 Aug;15(8):486-99. (PMID: 26205583) Sci Rep. 2017 Apr 7;7(1):737. (PMID: 28389661) Nature. 2019 Jul;571(7764):211-218. (PMID: 31207603) Nature. 2019 Dec;576(7786):293-300. (PMID: 31802004) Cell. 2023 Oct 12;186(21):4567-4582.e20. (PMID: 37794590) Nat Med. 2018 Jul;24(7):927-930. (PMID: 29892067) Nat Biomed Eng. 2024 Feb;8(2):177-192. (PMID: 37872368) Nat Genet. 2020 Jul;52(7):662-668. (PMID: 32424350) JCI Insight. 2017 Dec 7;2(23):. (PMID: 29212954) Nat Biotechnol. 2020 Jun;38(6):722-727. (PMID: 32518401) Nat Rev Immunol. 2019 Feb;19(2):73-74. (PMID: 30631206) Clin Cancer Res. 2017 May 1;23(9):2255-2266. (PMID: 27815355) Bioinformatics. 2009 Aug 15;25(16):2078-9. (PMID: 19505943) Nature. 2023 Mar;615(7952):507-516. (PMID: 36890224) Cell Rep. 2021 Dec 7;37(10):110083. (PMID: 34879274) Science. 2021 Apr 2;372(6537):. (PMID: 33795428) Cell Rep. 2018 Feb 6;22(6):1509-1521. (PMID: 29425506) Nat Immunol. 2019 Oct;20(10):1335-1347. (PMID: 31527834) Mol Cancer. 2022 Mar 18;21(1):78. (PMID: 35303871) Cell. 2018 Apr 19;173(3):665-676.e14. (PMID: 29551272) Proc Natl Acad Sci U S A. 2020 Jun 9;117(23):12969-12979. (PMID: 32434911) Cell. 2022 May 12;185(10):1745-1763.e22. (PMID: 35483375) Science. 2020 Feb 28;367(6481):. (PMID: 32029687) Nature. 2022 Mar;603(7903):934-941. (PMID: 35130560) Nature. 2023 Feb;614(7949):635-648. (PMID: 36813894) Nat Commun. 2019 Jun 11;10(1):2544. (PMID: 31186424) PLoS Pathog. 2015 Oct 20;11(10):e1005177. (PMID: 26485519) |
| 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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