Skip to main content

Advertisement

SpringerLink
Log in

CRISPR Takes the Front Seat in CART-Cell Development

  • Leading Article
  • Published:
BioDrugs Aims and scope Submit manuscript

Abstract

Chimeric antigen receptor T (CART)-cell immunotherapies have opened a door in the development of specialized gene therapies for hematological and solid cancers. Impressive response rates in pivotal trials led to the FDA approval of CART-cell therapy for certain hematological malignancies. However, autologous CART products are costly and time-intensive to manufacture, and most patients experience disease relapse within 1 year of CART administration. Additionally, CART-cell efficacy in solid tumors is extremely limited. CART-cell therapy is also associated with serious toxicities. Manufacturing difficulties, intrinsic T-cell defects, CART exhaustion, and treatment-associated toxicities are some of the current barriers to widespread adoption of CART-cell therapy. Genome editing tools such as CRISPR/Cas systems have demonstrated efficacy in further engineering CART cells to overcome these limitations. In this review, we will summarize the current approaches that use CRISPR to facilitate off-the-shelf CART products, increase CART-cell efficacy, and minimize CART-associated toxicities.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1

Similar content being viewed by others

References

  1. Mitsuyasu RT, Anton PA, Deeks SG, Scadden DT, Connick E, Downs MT, et al. Prolonged survival and tissue trafficking following adoptive transfer of CD4zeta gene-modified autologous CD4(+) and CD8(+) T cells in human immunodeficiency virus-infected subjects. Blood. 2000;96(3):785–93.

    Article  CAS  PubMed  Google Scholar 

  2. Walker RE, Bechtel CM, Natarajan V, Baseler M, Hege KM, Metcalf JA, et al. Long-term in vivo survival of receptor-modified syngeneic T cells in patients with human immunodeficiency virus infection. Blood. 2000;96(2):467–74.

    CAS  PubMed  Google Scholar 

  3. Kershaw MH, Westwood JA, Parker LL, Wang G, Eshhar Z, Mavroukakis SA, et al. A phase I study on adoptive immunotherapy using gene-modified T cells for ovarian cancer. Clin Cancer Res. 2006;12(20 Pt 1):6106–15.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Till BG, Jensen MC, Wang J, Chen EY, Wood BL, Greisman HA, et al. Adoptive immunotherapy for indolent non-Hodgkin lymphoma and mantle cell lymphoma using genetically modified autologous CD20-specific T cells. Blood. 2008;112(6):2261–71.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Moeller M, Haynes NM, Trapani JA, Teng MW, Jackson JT, Tanner JE, et al. A functional role for CD28 costimulation in tumor recognition by single-chain receptor-modified T cells. Cancer Gene Ther. 2004;11(5):371–9.

    Article  CAS  PubMed  Google Scholar 

  6. Savoldo B, Ramos CA, Liu E, Mims MP, Keating MJ, Carrum G, et al. CD28 costimulation improves expansion and persistence of chimeric antigen receptor-modified T cells in lymphoma patients. J Clin Invest. 2011;121(5):1822–6.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Milone MC, Fish JD, Carpenito C, Carroll RG, Binder GK, Teachey D, et al. Chimeric receptors containing CD137 signal transduction domains mediate enhanced survival of T cells and increased antileukemic efficacy in vivo. Mol Ther. 2009;17(8):1453–64.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Porter DL, Hwang WT, Frey NV, Lacey SF, Shaw PA, Loren AW, et al. Chimeric antigen receptor T cells persist and induce sustained remissions in relapsed refractory chronic lymphocytic leukemia. Sci Transl Med. 2015;7(303):303–139.

    Article  Google Scholar 

  9. Maude SL, Laetsch TW, Buechner J, Rives S, Boyer M, Bittencourt H, et al. Tisagenlecleucel in children and young adults with B-cell lymphoblastic leukemia. n Engl J Med. 2018;378(5):439–48.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Schuster SJ, Bishop MR, Tam CS, Waller EK, Borchmann P, McGuirk JP, et al. Tisagenlecleucel in adult relapsed or refractory diffuse large B-Cell lymphoma. N Engl J Med. 2019;380(1):45–56.

    Article  CAS  PubMed  Google Scholar 

  11. Neelapu SS, Locke FL, Bartlett NL, Lekakis LJ, Miklos DB, Jacobson CA, et al. Axicabtagene ciloleucel CAR T-cell therapy in refractory large B-Cell lymphoma. New Engl J Med. 2017;377(26):2531–44.

    Article  CAS  PubMed  Google Scholar 

  12. Wang M, Munoz J, Goy A, Locke FL, Jacobson CA, Hill BT, et al. KTE-X19 CAR T-cell therapy in relapsed or refractory mantle-cell lymphoma. N Engl J Med. 2020;382(14):1331–42.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Locke FL, Ghobadi A, Jacobson CA, Miklos DB, Lekakis LJ, Oluwole OO, et al. Long-term safety and activity of axicabtagene ciloleucel in refractory large B-cell lymphoma (ZUMA-1): a single-arm, multicentre, phase 1–2 trial. Lancet Oncol. 2019;20(1):31–42.

    Article  CAS  PubMed  Google Scholar 

  14. Majzner RG, Mackall CL. Clinical lessons learned from the first leg of the CAR T cell journey. Nat Med. 2019;25(9):1341–55.

    Article  CAS  PubMed  Google Scholar 

  15. Zhang H, Ye ZL, Yuan ZG, Luo ZQ, Jin HJ, Qian QJ. New Strategies for the Treatment of Solid Tumors with CAR-T Cells. Int J Biol Sci. 2016;12(6):718–29.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Urnov FD, Rebar EJ, Holmes MC, Zhang HS, Gregory PD. Genome editing with engineered zinc finger nucleases. Nat Rev Genet. 2010;11(9):636–46.

    Article  CAS  PubMed  Google Scholar 

  17. Sander JD, Joung JK. CRISPR-Cas systems for editing, regulating and targeting genomes. Nat Biotechnol. 2014;32(4):347–55.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Miller JC, Tan S, Qiao G, Barlow KA, Wang J, Xia DF, et al. A TALE nuclease architecture for efficient genome editing. Nat Biotechnol. 2011;29(2):143–8.

    Article  CAS  PubMed  Google Scholar 

  19. Gaj T, Gersbach CA, Barbas CF 3rd. ZFN, TALEN, and CRISPR/Cas-based methods for genome engineering. Trends Biotechnol. 2013;31(7):397–405.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Paques F, Duchateau P. Meganucleases and DNA double-strand break-induced recombination: perspectives for gene therapy. Curr Gene Ther. 2007;7(1):49–66.

    Article  CAS  PubMed  Google Scholar 

  21. Singh N, Shi J, June CH, Ruella M. Genome-editing technologies in adoptive T cell immunotherapy for cancer. Curr Hematol Malig Rep. 2017;12(6):522–9.

    Article  PubMed  PubMed Central  Google Scholar 

  22. Cong L, Ran FA, Cox D, Lin S, Barretto R, Habib N, et al. Multiplex genome engineering using CRISPR/Cas systems. Science. 2013;339(6121):819–23.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Ren J, Zhang X, Liu X, Fang C, Jiang S, June CH, et al. A versatile system for rapid multiplex genome-edited CAR T cell generation. Oncotarget. 2017;8(10):17002–11.

    Article  PubMed  PubMed Central  Google Scholar 

  24. Liu X, Zhang Y, Cheng C, Cheng AW, Zhang X, Li N, et al. CRISPR-Cas9-mediated multiplex gene editing in CAR-T cells. Cell Res. 2017;27(1):154–7.

    Article  PubMed  Google Scholar 

  25. Eyquem J, Mansilla-Soto J, Giavridis T, van der Stegen SJ, Hamieh M, Cunanan KM, et al. Targeting a CAR to the TRAC locus with CRISPR/Cas9 enhances tumour rejection. Nature. 2017;543(7643):113–7.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Fraietta JA, Nobles CL, Sammons MA, Lundh S, Carty SA, Reich TJ, et al. Disruption of TET2 promotes the therapeutic efficacy of CD19-targeted T cells. Nature. 2018;558(7709):307–12.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Ishino Y, Shinagawa H, Makino K, Amemura M, Nakata A. Nucleotide sequence of the iap gene, responsible for alkaline phosphatase isozyme conversion in Escherichia coli, and identification of the gene product. J Bacteriol. 1987;169(12):5429–33.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Bolotin A, Quinquis B, Sorokin A, Ehrlich SD. Clustered regularly interspaced short palindrome repeats (CRISPRs) have spacers of extrachromosomal origin. Microbiol (Reading). 2005;151(Pt 8):2551–61.

    Article  CAS  Google Scholar 

  29. Mojica FJ, Diez-Villasenor C, Garcia-Martinez J, Soria E. Intervening sequences of regularly spaced prokaryotic repeats derive from foreign genetic elements. J Mol Evol. 2005;60(2):174–82.

    Article  CAS  PubMed  Google Scholar 

  30. Brouns SJ, Jore MM, Lundgren M, Westra ER, Slijkhuis RJ, Snijders AP, et al. Small CRISPR RNAs guide antiviral defense in prokaryotes. Science. 2008;321(5891):960–4.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Barrangou R, Fremaux C, Deveau H, Richards M, Boyaval P, Moineau S, et al. CRISPR provides acquired resistance against viruses in prokaryotes. Science. 2007;315(5819):1709–12.

    Article  CAS  PubMed  Google Scholar 

  32. Deltcheva E, Chylinski K, Sharma CM, Gonzales K, Chao Y, Pirzada ZA, et al. CRISPR RNA maturation by trans-encoded small RNA and host factor RNase III. Nature. 2011;471(7340):602–7.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  33. Gasiunas G, Barrangou R, Horvath P, Siksnys V. Cas9-crRNA ribonucleoprotein complex mediates specific DNA cleavage for adaptive immunity in bacteria. Proc Natl Acad Sci USA. 2012;109(39):E2579–86.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Jinek M, Chylinski K, Fonfara I, Hauer M, Doudna JA, Charpentier E. A programmable dual-RNA-guided DNA endonuclease in adaptive bacterial immunity. Science. 2012;337(6096):816–21.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  35. Jinek M, East A, Cheng A, Lin S, Ma E, Doudna J. RNA-programmed genome editing in human cells. Elife. 2013;2:e00471.

    Article  PubMed  PubMed Central  Google Scholar 

  36. Mali P, Yang L, Esvelt KM, Aach J, Guell M, DiCarlo JE, et al. RNA-guided human genome engineering via Cas9. Science. 2013;339(6121):823–6.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  37. Zetsche B, Gootenberg JS, Abudayyeh OO, Slaymaker IM, Makarova KS, Essletzbichler P, et al. Cpf1 is a single RNA-guided endonuclease of a class 2 CRISPR-Cas system. Cell. 2015;163(3):759–71.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  38. Zetsche B, Heidenreich M, Mohanraju P, Fedorova I, Kneppers J, DeGennaro EM, et al. Multiplex gene editing by CRISPR-Cpf1 using a single crRNA array. Nat Biotechnol. 2017;35(1):31–4.

    Article  CAS  PubMed  Google Scholar 

  39. Batra SA, Rathi P, Guo L, Courtney AN, Fleurence J, Balzeau J, et al. Glypican-3-specific CAR T cells coexpressing IL15 and IL21 have superior expansion and antitumor activity against hepatocellular carcinoma. Cancer Immunol Res. 2020;8(3):309–20.

    Article  CAS  PubMed  Google Scholar 

  40. Choi BD, Yu X, Castano AP, Darr H, Henderson DB, Bouffard AA, et al. CRISPR-Cas9 disruption of PD-1 enhances activity of universal EGFRvIII CAR T cells in a preclinical model of human glioblastoma. J Immunother Cancer. 2019;7(1):304.

    Article  PubMed  PubMed Central  Google Scholar 

  41. Cooper ML, Choi J, Staser K, Ritchey JK, Devenport JM, Eckardt K, et al. An “off-the-shelf” fratricide-resistant CAR-T for the treatment of T cell hematologic malignancies. Leukemia. 2018;32(9):1970–83.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  42. Cox MJ, Kuhlmann CJ, Sterner RM, Sakemura R, Sinha S, Mwangi R, et al. Improved anti-tumor response of chimeric antigen receptor T cell (CART) therapy after GM-csf inhibition is mechanistically supported by a novel direct interaction of GM-CSF with activated carts. Blood. 2019;134(1):3868.

    Article  Google Scholar 

  43. Dai X, Park JJ, Du Y, Kim HR, Wang G, Errami Y, et al. One-step generation of modular CAR-T cells with AAV-Cpf1. Nat Methods. 2019;16(3):247–54.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  44. Li C, Mei H, Hu Y. Applications and explorations of CRISPR/Cas9 in CAR T-cell therapy. Brief Funct Genomics. 2020;19(3):175–82.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  45. Ren J, Liu X, Fang C, Jiang S, June CH, Zhao Y. Multiplex genome editing to generate universal CAR T cells resistant to PD1 inhibition. Clin Cancer Res. 2017;23(9):2255–66.

    Article  CAS  PubMed  Google Scholar 

  46. Siegler EL, Simone BW, Sakemura R, Tapper EE, Cox M, Horvei P, et al. Improving CART cell therapy with CRISPR-Cas12a gene editing for more efficient PD-1 gene knockout. Genome Writers Guilt. 2020;2020:19.

    Google Scholar 

  47. Stadtmauer EA, Cohen AD, Weber K, Lacey SF, Gonzalez VE, Melenhorst JJ, et al. First-in-human assessment of feasibility and safety of multiplexed genetic engineering of autologous T cells expressing NY-ESO -1 TCR and CRISPR/Cas9 gene edited to eliminate endogenous TCR and PD-1 (NYCE T cells) in advanced multiple myeloma (MM) and sarcoma. Blood. 2019;134(1):49.

    Article  Google Scholar 

  48. Stadtmauer EA, Fraietta JA, Davis MM, Cohen AD, Weber KL, Lancaster E, et al. CRISPR-engineered T cells in patients with refractory cancer. Science. 2020;367:6481.

    Article  Google Scholar 

  49. Sterner RM, Sakemura R, Cox MJ, Yang N, Khadka RH, Forsman CL, et al. GM-CSF inhibition reduces cytokine release syndrome and neuroinflammation but enhances CAR-T cell function in xenografts. Blood. 2018;2018:5.

    Google Scholar 

  50. Kouranova E, Forbes K, Zhao G, Warren J, Bartels A, Wu Y, et al. CRISPRs for optimal targeting: delivery of CRISPR components as DNA, RNA, and protein into cultured cells and single-cell embryos. Hum Gene Ther. 2016;27(6):464–75.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  51. Fajrial AK, He QQ, Wirusanti NI, Slansky JE, Ding X. A review of emerging physical transfection methods for CRISPR/Cas9-mediated gene editing. Theranostics. 2020;10(12):5532–49.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  52. Yu X, Liang X, Xie H, Kumar S, Ravinder N, Potter J, et al. Improved delivery of Cas9 protein/gRNA complexes using lipofectamine CRISPRMAX. Biotechnol Lett. 2016;38(6):919–29.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  53. Ruella M, Kenderian SS. Next-generation chimeric antigen receptor T-cell therapy: going off the shelf. BioDrugs. 2017;31(6):473–81.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  54. Nianias A, Themeli M. Induced pluripotent stem cell (iPSC)-derived lymphocytes for adoptive cell immunotherapy: recent advances and challenges. Curr Hematol Malig Rep. 2019;14(4):261–8.

    Article  PubMed  PubMed Central  Google Scholar 

  55. McEwan B, Padalia Z, Porras A, Sagert J, Terrett JA, Ho T, et al. Abstract 1428: allogeneic CRISPR/Cas9 gene-edited CAR-T cells targeting CD33 show potent preclinical activity against AML cells. Cancer Res. 2019;79(13):1428.

    Article  Google Scholar 

  56. Torikai H, Reik A, Soldner F, Warren EH, Yuen C, Zhou Y, et al. Toward eliminating HLA class I expression to generate universal cells from allogeneic donors. Blood. 2013;122(8):1341–9.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  57. Choi EI, Reimann KA, Letvin NL. In vivo natural killer cell depletion during primary simian immunodeficiency virus infection in rhesus monkeys. J Virol. 2008;82(13):6758–61.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  58. Roth TL, Puig-Saus C, Yu R, Shifrut E, Carnevale J, Li PJ, et al. Reprogramming human T cell function and specificity with non-viral genome targeting. Nature. 2018;559(7714):405–9.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  59. Themeli M, Kloss CC, Ciriello G, Fedorov VD, Perna F, Gonen M, et al. Generation of tumor-targeted human T lymphocytes from induced pluripotent stem cells for cancer therapy. Nat Biotechnol. 2013;31(10):928–33.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  60. Chang C, Van Der Stegen S, Mili M, Clarke R, Lai Y-S, Witty A, et al. FT819: translation of off-the-shelf TCR-Less Trac-1XX CAR-T cells in support of first-of-kind phase I clinical trial. Blood. 2019;134(1):4434.

    Article  Google Scholar 

  61. Ode Z, Condori J, Peterson N, Zhou S, Krenciute G. CRISPR-mediated non-viral site-specific gene integration and expression in T cells: protocol and application for T-Cell therapy. Cancers (Basel). 2020;12:6.

    Article  Google Scholar 

  62. Roth TL, Li PJ, Blaeschke F, Nies JF, Apathy R, Mowery C, et al. Pooled knockin targeting for genome engineering of cellular immunotherapies. Cell. 2020;181(3):728–44.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  63. MacLeod DT, Antony J, Martin AJ, Moser RJ, Hekele A, Wetzel KJ, et al. Integration of a CD19 CAR into the TCR alpha chain locus streamlines production of allogeneic gene-edited CAR T cells. Mol Ther. 2017;25(4):949–61.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  64. Martinez M, Moon EK. CAR T cells for solid tumors: new strategies for finding, infiltrating, and surviving in the tumor microenvironment. Front Immunol. 2019;10:128.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  65. Poorebrahim M, Melief J, de Coana YP, Wickstrom SL, Cid-Arregui A, Kiessling R. Counteracting CAR T cell dysfunction. Oncogene. 2020;2020:8.

    Google Scholar 

  66. Tschumi BO, Dumauthioz N, Marti B, Zhang L, Lanitis E, Irving M, et al. CART cells are prone to Fas- and DR5-mediated cell death. J Immunother Cancer. 2018;6(1):71.

    Article  PubMed  PubMed Central  Google Scholar 

  67. Grosser R, Cherkassky L, Chintala N, Adusumilli PS. Combination immunotherapy with CAR T cells and checkpoint blockade for the treatment of solid tumors. Cancer Cell. 2019;36(5):471–82.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  68. Rupp LJ, Schumann K, Roybal KT, Gate RE, Ye CJ, Lim WA, et al. CRISPR/Cas9-mediated PD-1 disruption enhances anti-tumor efficacy of human chimeric antigen receptor T cells. Sci Rep. 2017;7(1):737.

    Article  PubMed  PubMed Central  Google Scholar 

  69. Galon J, Rossi J, Turcan S, Danan C, Locke FL, Neelapu SS, et al. Characterization of anti-CD19 chimeric antigen receptor (CAR) T cell-mediated tumor microenvironment immune gene profile in a multicenter trial (ZUMA-1) with axicabtagene ciloleucel (axi-cel, KTE-C19). J Clin Oncol. 2017;35(15):3025.

    Article  Google Scholar 

  70. Zhang Y, Zhang X, Cheng C, Mu W, Liu X, Li N, et al. CRISPR-Cas9 mediated LAG-3 disruption in CAR-T cells. Front Med. 2017;11(4):554–62.

    Article  PubMed  Google Scholar 

  71. Tang N, Cheng C, Zhang X, Qiao M, Li N, Mu W, et al. TGF-beta inhibition via CRISPR promotes the long-term efficacy of CAR T cells against solid tumors. JCI Insight. 2020;5:4.

    Article  Google Scholar 

  72. Waring P, Mullbacher A. Cell death induced by the Fas/Fas ligand pathway and its role in pathology. Immunol Cell Biol. 1999;77(4):312–7.

    Article  CAS  PubMed  Google Scholar 

  73. Krammer PH. CD95’s deadly mission in the immune system. Nature. 2000;407(6805):789–95.

    Article  CAS  PubMed  Google Scholar 

  74. Chen X, Zachar V, Zdravkovic M, Guo M, Ebbesen P, Liu X. Role of the Fas/Fas ligand pathway in apoptotic cell death induced by the human T cell lymphotropic virus type I Tax transactivator. J Gen Virol. 1997;78(Pt 12):3277–85.

    Article  CAS  PubMed  Google Scholar 

  75. Kunkele A, Johnson AJ, Rolczynski LS, Chang CA, Hoglund V, Kelly-Spratt KS, et al. Functional tuning of CARs reveals signaling threshold above which CD8+ ctl antitumor potency is attenuated due to cell Fas-FasL-dependent AICD. Cancer Immunol Res. 2015;3(4):368–79.

    Article  PubMed  Google Scholar 

  76. Lynn RC, Weber EW, Sotillo E, Gennert D, Xu P, Good Z, et al. c-Jun overexpression in CAR T cells induces exhaustion resistance. Nature. 2019;576(7786):293–300.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  77. Petersen C, Bell M, Houke H, Yi Z, Gottschalk S, Krenciute G. Immu-13. CRISPR/CAS9-mediated silencing of SHP-1 significantly enhances the anti-glioma activity of IL-13Rα2 CAR T cells. Neuro-Oncol. 2019;21(2):95–6.

  78. Siegler EL, Kenderian SS. Neurotoxicity and cytokine release syndrome after chimeric antigen receptor T cell therapy: insights into mechanisms and novel therapies. Front Immunol. 2020;11:1973.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  79. Ruff MW, Siegler EL, Kenderian SS. A concise review of neurologic complications associated with chimeric antigen receptor T-cell immunotherapy. Neurol Clin. 2020;38(4):953–63.

    Article  PubMed  Google Scholar 

  80. Le RQ, Li L, Yuan W, Shord SS, Nie L, Habtemariam BA, et al. FDA approval summary: tocilizumab for treatment of chimeric antigen receptor T cell-induced severe or life-threatening cytokine release syndrome. Oncologist. 2018;23(8):943–7.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  81. Morgan RA, Yang JC, Kitano M, Dudley ME, Laurencot CM, Rosenberg SA. Case report of a serious adverse event following the administration of T cells transduced with a chimeric antigen receptor recognizing ERBB2. Mol Ther. 2010;18(4):843–51.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  82. Sterner RM, Sakemura R, Cox MJ, Yang N, Khadka RH, Forsman CL, et al. GM-CSF inhibition reduces cytokine release syndrome and neuroinflammation but enhances CAR-T cell function in xenografts. Blood. 2019;133(7):697–709.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  83. Sterner RM, Cox MJ, Sakemura R, Kenderian SS. Using CRISPR/Cas9 to Knock Out GM-CSF in CAR-T Cells. J Vis Exp. 2019;2019:149.

    Google Scholar 

  84. Hu B, Yi Y, Wang X, Tao G. 487 Automatic neutralization of IL6 storm and blockade of IL1 signaling during CART therapy to reduce cytokine toxicity and minimize neurotoxicity. Mol Ther. 2020;28:4S1.

    Google Scholar 

  85. Kim MY, Kenderian SS, Schreeder D, Klichinsky M, Kozlowski M, Schestova O, et al. Genome editing using CRISPR-Cas9 to increase the therapeutic index of antigen-specific immunotherapy in acute myeloid leukemia. Mol Therapy. 2016;24:S108–9.

    Article  Google Scholar 

  86. Campana D, Behm FG. Immunophenotyping of leukemia. J Immunol Methods. 2000;243(1–2):59–75.

    Article  CAS  PubMed  Google Scholar 

  87. Milush JM, Long BR, Snyder-Cappione JE, Cappione AJ 3rd, York VA, Ndhlovu LC, et al. Functionally distinct subsets of human NK cells and monocyte/DC-like cells identified by coexpression of CD56, CD7, and CD4. Blood. 2009;114(23):4823–31.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  88. Gomes-Silva D, Srinivasan M, Sharma S, Lee CM, Wagner DL, Davis TH, et al. CD7-edited T cells expressing a CD7-specific CAR for the therapy of T-cell malignancies. Blood. 2017;130(3):285–96.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  89. Raikar SS, Fleischer LC, Moot R, Fedanov A, Paik NY, Knight KA, et al. Development of chimeric antigen receptors targeting T-cell malignancies using two structurally different anti-CD5 antigen binding domains in NK and CRISPR-edited T cell lines. Oncoimmunology. 2018;7(3):e1407898.

    Article  PubMed  Google Scholar 

  90. Peng R, Lin G, Li J. Potential pitfalls of CRISPR/Cas9-mediated genome editing. FEBS J. 2016;283(7):1218–31.

    Article  CAS  PubMed  Google Scholar 

  91. Fu Y, Foden JA, Khayter C, Maeder ML, Reyon D, Joung JK, et al. High-frequency off-target mutagenesis induced by CRISPR-Cas nucleases in human cells. Nat Biotechnol. 2013;31(9):822–6.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  92. Naeem M, Majeed S, Hoque MZ, Ahmad I. Latest developed strategies to minimize the off-target effects in CRISPR-Cas-mediated genome editing. Cells. 2020;9:7.

    Article  Google Scholar 

  93. Wong N, Liu W, Wang X. WU-CRISPR: characteristics of functional guide RNAs for the CRISPR/Cas9 system. Genome Biol. 2015;16:218.

    Article  PubMed  PubMed Central  Google Scholar 

  94. Fu Y, Sander JD, Reyon D, Cascio VM, Joung JK. Improving CRISPR-Cas nuclease specificity using truncated guide RNAs. Nat Biotechnol. 2014;32(3):279–84.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  95. Ryan DE, Taussig D, Steinfeld I, Phadnis SM, Lunstad BD, Singh M, et al. Improving CRISPR-Cas specificity with chemical modifications in single-guide RNAs. Nucleic Acids Res. 2018;46(2):792–803.

    Article  CAS  PubMed  Google Scholar 

  96. Wang T, Wei JJ, Sabatini DM, Lander ES. Genetic screens in human cells using the CRISPR-Cas9 system. Science. 2014;343(6166):80–4.

    Article  CAS  PubMed  Google Scholar 

  97. Prykhozhij SV, Rajan V, Gaston D, Berman JN. Correction: CRISPR MultiTargeter: a web tool to find common and unique CRISPR single guide RNA targets in a set of similar sequences. PLoS ONE. 2015;10(9):e0138634.

    Article  PubMed  PubMed Central  Google Scholar 

  98. Bae S, Park J, Kim JS. Cas-OFFinder: a fast and versatile algorithm that searches for potential off-target sites of Cas9 RNA-guided endonucleases. Bioinformatics. 2014;30(10):1473–5.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  99. O’Brien A, Bailey TL. GT-Scan: identifying unique genomic targets. Bioinformatics. 2014;30(18):2673–5.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  100. Friedland AE, Baral R, Singhal P, Loveluck K, Shen S, Sanchez M, et al. Characterization of Staphylococcus aureus Cas9: a smaller Cas9 for all-in-one adeno-associated virus delivery and paired nickase applications. Genome Biol. 2015;16:257.

    Article  PubMed  PubMed Central  Google Scholar 

  101. Rahdar M, McMahon MA, Prakash TP, Swayze EE, Bennett CF, Cleveland DW. Synthetic CRISPR RNA-Cas9-guided genome editing in human cells. Proc Natl Acad Sci USA. 2015;112(51):E7110–7.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  102. Kleinstiver BP, Prew MS, Tsai SQ, Topkar VV, Nguyen NT, Zheng Z, et al. Engineered CRISPR-Cas9 nucleases with altered PAM specificities. Nature. 2015;523(7561):481–5.

    Article  PubMed  PubMed Central  Google Scholar 

  103. Kleinstiver BP, Tsai SQ, Prew MS, Nguyen NT, Welch MM, Lopez JM, et al. Genome-wide specificities of CRISPR-Cas Cpf1 nucleases in human cells. Nat Biotechnol. 2016;34(8):869–74.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  104. Charlesworth CT, Deshpande PS, Dever DP, Camarena J, Lemgart VT, Cromer MK, et al. Identification of preexisting adaptive immunity to Cas9 proteins in humans. Nature Med. 2019;25(2):249.

    Article  CAS  PubMed  Google Scholar 

  105. Ferdosi SR, Ewaisha R, Moghadam F, Krishna S, Park JG, Ebrahimkhani MR, et al. Multifunctional CRISPR-Cas9 with engineered immunosilenced human T cell epitopes. Nat Commun. 2019;10(1):1842.

    Article  PubMed  PubMed Central  Google Scholar 

  106. Harrington LB, Paez-Espino D, Staahl BT, Chen JS, Ma E, Kyrpides NC, et al. A thermostable Cas9 with increased lifetime in human plasma. Nat Commun. 2017;8(1):1424.

    Article  PubMed  PubMed Central  Google Scholar 

  107. Kim S, Koo T, Jee HG, Cho HY, Lee G, Lim DG, et al. CRISPR RNAs trigger innate immune responses in human cells. Genome Res. 2018;2018:6.

    Google Scholar 

  108. Athanasopoulos T, Munye MM, Yanez-Munoz RJ. Nonintegrating gene therapy vectors. Hematol Oncol Clin N Am. 2017;31(5):753–70.

    Article  Google Scholar 

  109. Bobis-Wozowicz S, Galla M, Alzubi J, Kuehle J, Baum C, Schambach A, et al. Non-integrating gamma-retroviral vectors as a versatile tool for transient zinc-finger nuclease delivery. Sci Rep. 2014;4:4656.

    Article  PubMed  PubMed Central  Google Scholar 

  110. Xu X, Wan T, Xin H, Li D, Pan H, Wu J, et al. Delivery of CRISPR/Cas9 for therapeutic genome editing. J Gene Med. 2019;21(7):e3107.

    Article  PubMed  Google Scholar 

  111. Lao YH, Li M, Gao MA, Shao D, Chi CW, Huang D, et al. HPV oncogene manipulation using nonvirally delivered CRISPR/Cas9 or Natronobacterium gregoryi argonaute. Adv Sci (Weinh). 2018;5(7):1700540.

    Article  PubMed  PubMed Central  Google Scholar 

  112. Wan T, Niu D, Wu CB, Xu FJ, Church G, Ping Y. Material solutions for delivery of CRISPR/Cas-based genome editing tools: current status and future outlook. Mater Today. 2019;26:40–66.

    Article  CAS  Google Scholar 

  113. Lee K, Conboy M, Park HM, Jiang F, Kim HJ, Dewitt MA, et al. Nanoparticle delivery of Cas9 ribonucleoprotein and donor DNA in vivo induces homology-directed DNA repair. Nat Biomed Eng. 2017;1:889–901.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  114. Kosicki M, Tomberg K, Bradley A. Repair of double-strand breaks induced by CRISPR-Cas9 leads to large deletions and complex rearrangements (vol 36, pg 765, 2018). Nature Biotechnol. 2018;36(9):899.

  115. Chu VT, Weber T, Wefers B, Wurst W, Sander S, Rajewsky K, et al. Increasing the efficiency of homology-directed repair for CRISPR-Cas9-induced precise gene editing in mammalian cells. Nat Biotechnol. 2015;33(5):543-U160.

    Article  CAS  PubMed  Google Scholar 

  116. Maruyama T, Dougan SK, Truttmann MC, Bilate AM, Ingram JR, Ploegh HL. Increasing the efficiency of precise genome editing with CRISPR-Cas9 by inhibition of nonhomologous end joining. Nat Biotechnol. 2015;33(5):538–42.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  117. Lin S, Staahl B, Alla RK, Doudna JA. Enhanced homology-directed human genome engineering by controlled timing of CRISPR/Cas9 delivery. Elife. 2014;2014:3.

    Google Scholar 

  118. Tsai SQ, Joung JK. Defining and improving the genome-wide specificities of CRISPR-Cas9 nucleases. Nat Rev Genet. 2016;17(5):300–12.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  119. Kim D, Bae S, Park J, Kim E, Kim S, Yu HR, et al. Digenome-seq: genome-wide profiling of CRISPR-Cas9 off-target effects in human cells. Nat Methods. 2015;12(3):237–43.

    Article  CAS  PubMed  Google Scholar 

  120. Kim MY, Yu KR, Kenderian SS, Ruella M, Chen S, Shin TH, et al. Genetic inactivation of CD33 in hematopoietic stem cells to enable CAR T cell immunotherapy for acute myeloid leukemia. Cell. 2018;173(6):1439–53.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. T Cell Engineering, Mayo Clinic, Rochester, MN, USA

    Claudia Manriquez-Roman, Elizabeth L. Siegler & Saad S. Kenderian

  2. Division of Hematology, Mayo Clinic, 200 First Street SW, Rochester, MN, 55905, USA

    Claudia Manriquez-Roman, Elizabeth L. Siegler & Saad S. Kenderian

  3. Department of Immunology, Mayo Clinic, Rochester, MN, USA

    Saad S. Kenderian

  4. Department of Molecular Medicine, Mayo Clinic, Rochester, MN, USA

    Claudia Manriquez-Roman & Saad S. Kenderian

  5. Mayo Clinic Graduate School of Biomedical Sciences, Mayo Clinic, Rochester, MN, USA

    Claudia Manriquez-Roman & Saad S. Kenderian

Authors
  1. Claudia Manriquez-Roman
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Elizabeth L. Siegler
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Saad S. Kenderian
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Saad S. Kenderian.

Ethics declarations

Funding

This work was partly supported through the Mayo Clinic K2R pipeline (SSK), the Mayo Clinic Center for Individualized Medicine (SSK), and the Predolin Foundation (SSK).

Disclosures

SSK is an inventor on patents in the field of CAR immunotherapy that are licensed to Novartis (through an agreement between Mayo Clinic, University of Pennsylvania, and Novartis), to Mettaforge (through Mayo Clinic), and to Humanigen (through Mayo Clinic). SSK receives research funding from Kite, Gilead, Juno, Celgene, Novartis, Humanigen, MorphoSys, Tolero, Sunesis, Leahlabs, and Lentigen. These disclosures are not directly related to the content of this manuscript. CMR and ELS declare that they have no conflicts of interest that might be relevant to the contents of this manuscript.

Ethics approval

Not applicable.

Consent to participate

Not applicable.

Consent for publication

Not applicable.

Availability of data and material

Not applicable.

Code availability

Not applicable.

Authors' contributions

CMR, ELS, and SSK formulated the content of this review, wrote the manuscript, and approved the final version.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Manriquez-Roman, C., Siegler, E.L. & Kenderian, S.S. CRISPR Takes the Front Seat in CART-Cell Development. BioDrugs 35, 113–124 (2021). https://doi.org/10.1007/s40259-021-00473-y

Download citation

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s40259-021-00473-y

Search

Navigation