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Potent CAR-T cells engineered with Sleeping Beauty transposon vectors display a central memory phenotype

Gene Therapy volume 28, pages 3–5 (2021)Cite this article

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The Original Article was published on 09 January 2020

Several clinical studies have demonstrated that human T lymphocytes that express a chimeric antigen receptor (CAR) against the B-cell lineage marker CD19 (CD19+ CAR-T cells) are able to induce durable complete remissions in patients with chemotherapy- and radiotherapy-refractory B-cell acute lymphoblastic leukemia (ALL), non-Hodgkin lymphoma (NHL), and chronic lymphocytic leukemia [1,2,3,4,5,6,7,8]. Two CAR-T cell products have obtained marketing authorization in the United States and in the European Union for the treatment of CD19+ ALL and NHL, and both made headlines due to their considerable market price and the complex logistics behind this treatment.

There are several hundred entries that pop up on ClinicalTrials.gov by using “CAR-T” as query term. Indeed, clinical trials targeting a wide range of antigens for hematologic and solid tumors are currently under development; thus, it is predicted that the demand for CAR-T cell therapies will increase in the near future. The current CAR-T therapy supply chain involves harvesting the patient’s T cells at a leukapheresis center, shipping to a centralized manufacturing facility to perform CAR gene transfer and T-cell expansion, and return shipment of the cryopreserved cell product to the hospital, where the therapy is administered. This procedure undoubtedly represents complex logistics associated with significant costs and waiting times. In addition, the manufacturing capacities of specialized GMP production facilities will likely be exhausted, especially if CAR-T therapies will become available for more common forms of cancer affecting larger patient populations. Thus, in order to address the significant and foreseeable demand for CAR-T therapies in the near future [9] and to enable broad access to this therapy, manufacturing protocols that are more suitable to reduce production time and costs of the cell product are currently under development. There are at least three ongoing developments addressing the above needs. First, there is a recent increase in the use of exportable and automated manufacturing devices that are anticipated to provide decentralized, on-site, point-of-care CAR-T cell manufacture to reduce cost and waiting time [10, 11]. Second, recent preclinical evidence indicates the feasibility of carrying out CAR gene transfer directly in vivo, with the use of lentiviral vectors and nonviral nanocarriers specifically targeted to T lymphocytes [12, 13]. Such protocols carry the promise of completely bypassing ex vivo cell culture and associated quality control procedures. And third, novel technologies based on virus-free gene transfer (including transposons and the CRISPR/Cas system) can be applied to reduce time and costs of manufacturing clinical-grade CAR-T cells.

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References

  1. Turtle CJ, Hanafi LA, Berger C, Gooley TA, Cherian S, Hudecek M, et al. CD19 CAR-T cells of defined CD4+:CD8+ composition in adult B cell ALL patients. J Clin Investig. 2016;126:2123–38.

    Article  Google Scholar 

  2. Turtle CJ, Hanafi LA, Berger C, Hudecek M, Pender B, Robinson E, et al. Immunotherapy of non-Hodgkin’s lymphoma with a defined ratio of CD8+ and CD4+ CD19-specific chimeric antigen receptor-modified T cells. Sci Transl Med. 2016;8:355ra116.

    Article  Google Scholar 

  3. Turtle CJ, Maloney DG. Clinical trials of CD19-targeted CAR-modified T cell therapy; a complex and varied landscape. Expert Rev Hematol. 2016;9:719–21.

    Article  CAS  Google Scholar 

  4. Maude SL, Frey N, Shaw PA, Aplenc R, Barrett DM, Bunin NJ, et al. Chimeric antigen receptor T cells for sustained remissions in leukemia. N Engl J Med. 2014;371:1507–17.

    Article  Google Scholar 

  5. Davila ML, Riviere I, Wang X, Bartido S, Park J, Curran K, et al. Efficacy and toxicity management of 19-28z CAR T cell therapy in B cell acute lymphoblastic leukemia. Sci Transl Med. 2014;6:224–5.

    Article  Google Scholar 

  6. Lee DW, Kochenderfer JN, Stetler-Stevenson M, Cui YK, Delbrook C, Feldman SA, et al. T cells expressing CD19 chimeric antigen receptors for acute lymphoblastic leukaemia in children and young adults: a phase 1 dose-escalation trial. Lancet. 2015;385:517–28.

    Article  CAS  Google Scholar 

  7. Kochenderfer JN, Dudley ME, Kassim SH, Somerville RP, Carpenter RO, Stetler-Stevenson M, et al. Chemotherapy-refractory diffuse large B-cell lymphoma and indolent B-cell malignancies can be effectively treated with autologous T cells expressing an anti-CD19 chimeric antigen receptor. J Clin Oncol. 2015;33:540–9.

    Article  CAS  Google Scholar 

  8. Turtle CJ, Hay KA, Hanafi LA, Li D, Cherian S, Chen X, et al. Durable molecular remissions in chronic lymphocytic leukemia treated with CD19-specific chimeric antigen receptor-modified T cells after failure of ibrutinib. J Clin Oncol. 2017;35:3010–20.

    Article  CAS  Google Scholar 

  9. Hartmann J, Schussler-Lenz M, Bondanza A, Buchholz CJ. Clinical development of CAR T cells-challenges and opportunities in translating innovative treatment concepts. EMBO Mol Med. 2017;9:1183–97.

    Article  CAS  Google Scholar 

  10. Lock D, Mockel-Tenbrinck N, Drechsel K, Barth C, Mauer D, Schaser T, et al. Automated manufacturing of potent CD20-directed chimeric antigen receptor T cells for clinical use. Hum Gene Ther. 2017;28:914–25.

    Article  CAS  Google Scholar 

  11. Mock U, Nickolay L, Philip B, Cheung GW, Zhan H, Johnston IC, et al. Automated manufacturing of chimeric antigen receptor T cells for adoptive immunotherapy using CliniMACS prodigy. Cytotherapy. 2016;18:1002–11.

    Article  CAS  Google Scholar 

  12. Pfeiffer A, Thalheimer FB, Hartmann S, Frank AM, Bender RR, Danisch S, et al. In vivo generation of human CD19-CAR T cells results in B-cell depletion and signs of cytokine release syndrome. EMBO Mol Med. 2018;10:e9158.

    Article  Google Scholar 

  13. Smith TT, Stephan SB, Moffett HF, McKnight LE, Ji W, Reiman D, et al. In situ programming of leukaemia-specific T cells using synthetic DNA nanocarriers. Nat Nanotechnol. 2017;12:813–20.

    Article  CAS  Google Scholar 

  14. Chicaybam L, Abdo L, Viegas M, Marques LVC, de Sousa P, Batista-Silva LR, et al. Transposon-mediated generation of CAR-T cells shows efficient anti B-cell leukemia response after ex vivo expansion. Gene Ther. 2020;27:85–95.

    Article  CAS  Google Scholar 

  15. Hodge R, Narayanavari SA, Izsvak Z, Ivics Z. Wide awake and ready to move: 20 years of non-viral therapeutic genome engineering with the sleeping beauty transposon system. Hum Gene Ther. 2017;28:842–55.

    Article  CAS  Google Scholar 

  16. Hudecek M, Izsvak Z, Johnen S, Renner M, Thumann G, Ivics Z. Going non-viral: the Sleeping Beauty transposon system breaks on through to the clinical side. Crit Rev Biochem Mol Biol. 2017;52:355–80.

    Article  CAS  Google Scholar 

  17. Kebriaei P, Izsvak Z, Narayanavari SA, Singh H, Ivics Z. Gene therapy with the sleeping beauty transposon system. Trends Genet. 2017;33:852–70.

    Article  CAS  Google Scholar 

  18. Hackett PB, Largaespada DA, Cooper LJN. A transposon and transposase system for human application. Mol Ther. 2010;18:674–83.

    Article  CAS  Google Scholar 

  19. Zayed H, Izsvak Z, Walisko O, Ivics Z. Development of hyperactive sleeping beauty transposon vectors by mutational analysis. Mol Ther. 2004;9:292–304.

    Article  CAS  Google Scholar 

  20. Walisko O, Schorn A, Rolfs F, Devaraj A, Miskey C, Izsvak Z, et al. Transcriptional activities of the Sleeping Beauty transposon and shielding its genetic cargo with insulators. Mol Ther. 2008;16:359–69.

    Article  CAS  Google Scholar 

  21. Moldt B, Yant SR, Andersen PR, Kay MA, Mikkelsen JG. Cis-acting gene regulatory activities in the terminal regions of sleeping beauty DNA transposon-based vectors. Hum Gene Ther. 2007;18:1193–204.

    Article  CAS  Google Scholar 

  22. Ivics Z, Katzer A, Stuwe EE, Fiedler D, Knespel S, Izsvak Z. Targeted Sleeping Beauty transposition in human cells. Mol Ther. 2007;15:1137–44.

    Article  CAS  Google Scholar 

  23. VandenDriessche T, Ivics Z, Izsvak Z, Chuah MKL. Emerging potential of transposons for gene therapy and generation of induced pluripotent stem cells. Blood. 2009;114:1461–8.

    Article  CAS  Google Scholar 

  24. Yant SR, Meuse L, Chiu W, Ivics Z, Izsvak Z, Kay MA. Somatic integration and long-term transgene expression in normal and haemophilic mice using a DNA transposon system. Nat Genet. 2000;25:35–41.

    Article  CAS  Google Scholar 

  25. Mates L, Chuah MK, Belay E, Jerchow B, Manoj N, Acosta-Sanchez A, et al. Molecular evolution of a novel hyperactive Sleeping Beauty transposase enables robust stable gene transfer in vertebrates. Nat Genet. 2009;41:753–61.

    Article  CAS  Google Scholar 

  26. Chicaybam L, Abdo L, Carneiro M, Peixoto B, Viegas M, de Sousa P, et al. CAR T cells generated using Sleeping Beauty Transposon vectors and expanded with an EBV-transformed lymphoblastoid cell line display antitumor activity in vitro and in vivo. Hum Gene Ther. 2019;30:511–22.

    Article  CAS  Google Scholar 

  27. Huang X, Guo H, Kang J, Choi S, Zhou TC, Tammana S, et al. Sleeping Beauty transposon-mediated engineering of human primary T cells for therapy of CD19+ lymphoid malignancies. Mol Ther. 2008;16:580–9.

    Article  CAS  Google Scholar 

  28. Kawalekar OU, O’Connor RS, Fraietta JA, Guo L, McGettigan SE, Posey AD Jr., et al. Distinct signaling of coreceptors regulates specific metabolism pathways and impacts memory development in CAR T cells. Immunity. 2016;44:380–90.

    Article  CAS  Google Scholar 

  29. Morita D, Nishio N, Saito S, Tanaka M, Kawashima N, Okuno Y, et al. Enhanced expression of anti-CD19 chimeric antigen receptor in piggybac transposon-engineered T cells. Mol Ther. 2018;8:131–40.

    CAS  Google Scholar 

  30. Barnett BE, Hermanson DL, Smith JB, Wang X, Tan Y, Martin CE. piggyBac™-produced CAR-T cells exhibit stem-cell memory phenotype. Blood. 2016;128:2167.

    Article  Google Scholar 

  31. Sommermeyer D, Hudecek M, Kosasih PL, Gogishvili T, Maloney DG, Turtle CJ, et al. Chimeric antigen receptor-modified T cells derived from defined CD8+ and CD4+ subsets confer superior antitumor reactivity in vivo. Leukemia. 2016;30:492–500.

    Article  CAS  Google Scholar 

  32. Klebanoff CA, Gattinoni L, Torabi-Parizi P, Kerstann K, Cardones AR, Finkelstein SE, et al. Central memory self/tumor-reactive CD8+ T cells confer superior antitumor immunity compared with effector memory T cells. Proc Natl Acad Sci USA. 2005;102:9571–6.

    Article  CAS  Google Scholar 

  33. Monjezi R, Miskey C, Gogishvili T, Schleef M, Schmeer M, Einsele H, et al. Enhanced CAR T-cell engineering using non-viral Sleeping Beauty transposition from minicircle vectors. Leukemia. 2017;31:186–94.

    Article  CAS  Google Scholar 

  34. Kebriaei P, Singh H, Huls MH, Figliola MJ, Bassett R, Olivares S, et al. Phase I trials using Sleeping Beauty to generate CD19-specific CAR T cells. J Clin Investig. 2016;126:3363–76.

    Article  Google Scholar 

  35. Gogishvili T, Danhof S, Prommersberger S, Rydzek J, Schreder M, Brede C, et al. SLAMF7-CAR T cells eliminate myeloma and confer selective fratricide of SLAMF7(+) normal lymphocytes. Blood. 2017;130:2838–47.

    Article  CAS  Google Scholar 

  36. Hudecek M, Ivics Z. Non-viral therapeutic cell engineering with the Sleeping Beauty transposon system. Curr Opin Genet Dev. 2018;52:100–8.

    Article  CAS  Google Scholar 

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  1. Transposition and Genome Engineering, Division of Medical Biotechnology, Paul Ehrlich Institute, Paul Ehrlich Str. 51-59, D-63225, Langen, Germany

    Zoltán Ivics

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Correspondence to Zoltán Ivics.

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The author is an inventor of several patents relating to Sleeping Beauty transposon technology.

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Ivics, Z. Potent CAR-T cells engineered with Sleeping Beauty transposon vectors display a central memory phenotype. Gene Ther 28, 3–5 (2021). https://doi.org/10.1038/s41434-020-0138-8

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