Systematically optimized BCMA/CS1 bispecific CAR-T cells robustly control heterogeneous multiple myeloma
Eugenia Zah, Eunwoo Nam, Vinya Bhuvan, Uyen Tran, Brenda Y Ji, Stanley B Gosliner, Xiuli Wang, Christine E Brown, Yvonne Y Chen, Eugenia Zah, Eunwoo Nam, Vinya Bhuvan, Uyen Tran, Brenda Y Ji, Stanley B Gosliner, Xiuli Wang, Christine E Brown, Yvonne Y Chen
Abstract
Chimeric antigen receptor (CAR)-T cell therapy has shown remarkable clinical efficacy against B-cell malignancies, yet marked vulnerability to antigen escape and tumor relapse exists. Here we report the rational design and optimization of bispecific CAR-T cells with robust activity against heterogeneous multiple myeloma (MM) that is resistant to conventional CAR-T cell therapy targeting B-cell maturation antigen (BCMA). We demonstrate that BCMA/CS1 bispecific CAR-T cells exhibit superior CAR expression and function compared to T cells that co-express individual BCMA and CS1 CARs. Combination therapy with anti-PD-1 antibody further accelerates the rate of initial tumor clearance in vivo, while CAR-T cell treatment alone achieves durable tumor-free survival even upon tumor re-challenge. Taken together, the BCMA/CS1 bispecific CAR presents a promising treatment approach to prevent antigen escape in CAR-T cell therapy against MM, and the vertically integrated optimization process can be used to develop robust cell-based therapy against novel disease targets.
Conflict of interest statement
Y.Y.C. and E.Z. declare competing financial interest in the form of a patent application whose value may be affected by the publication of this work. The other authors declare no competing interests.
Figures
References
- Cancer Facts and Figures 2019, American Cancer Society (2019). .
- Cowan AJ, et al. Global burden of multiple myeloma. JAMA Oncol. 2018;98121:1221–1227. doi: 10.1001/jamaoncol.2018.2128.
- Moreau P, Attal M, Facon T. Frontline therapy of multiple myeloma. Blood. 2015;125:3076–3085. doi: 10.1182/blood-2014-09-568915.
- Ali SA, et al. T cells expressing an anti-B cell maturation antigen chimeric antigen receptor cause remissions of multiple myeloma. Blood. 2016;128:1688–1701. doi: 10.1182/blood-2016-04-711903.
- Brudno J, et al. T cells genetically modified to express an anti-B cell maturation antigen chimeric antigen receptor cause remissions of poor-prognosis relapsed multiple myeloma. J. Clin. Oncol. 2018;36:2267–2280. doi: 10.1200/JCO.2018.77.8084.
- Berdeja JG, et al. Durable clinical responses in heavily pretreated patients with relapsed/refractory multiple myeloma: Updated results from a multicenter study of bb2121 anti-Bcma CAR T cell therapy. Blood. 2017;130:740.
- Cohen AD, et al. Safety and efficacy of B cell maturation antigen (BCMA)-specific chimeric antigen receptor T cells (CART-BCMA) with cyclophosphamide conditioning for refractory multiple myeloma (MM) Blood. 2017;130:505. doi: 10.1182/blood-2017-08-803551.
- Smith EL, et al. Development and evaluation of a human single chain variable fragment (scFv) derived Bcma rargeted CAR T cell vector leads to a high objective response rate in patients with advanced MM. Blood. 2017;130:742. doi: 10.1182/blood-2017-02-769869.
- Grada Z, et al. TanCAR: A novel bispecific chimeric antigen receptor for cancer immunotherapy. Mol. Ther. Nucleic Acids. 2013;2:e105. doi: 10.1038/mtna.2013.32.
- Zah, E., Lin, M.-Y., Silva-Benedict, A., Jensen, M. C. & Chen, Y. Y. T cells expressing CD19/CD20 bi-specific chimeric antigen receptors prevent antigen escape by malignant B cells. Cancer Immunol. Res. 10.1158/2326-6066.CIR-15-0231 (2016).
- Hegde M, et al. Tandem CAR T cells targeting HER2 and IL13R α 2 mitigate tumor antigen escape. J. Clin. Invest. 2016;126:3036–3052. doi: 10.1172/JCI83416.
- Qin H, et al. Novel CD19/CD22 Bicistronic chimeric antigen receptors outperform single or bivalent CARs in eradicating CD19+CD22+, CD19−, and CD22− Pre-B Leukemia. Am. Soc. Hematol. 2017;130:810.
- Lee L, et al. An APRIL-based chimeric antigen receptor for dual targeting of BCMA and TACI in multiple myeloma. Blood. 2018;131:746–758. doi: 10.1182/blood-2017-05-781351.
- Novak AJ, et al. Expression of BCMA, TACI, and BAFF-R in multiple myeloma: a mechanism for growth and survival. Blood. 2004;103:689–694. doi: 10.1182/blood-2003-06-2043.
- Moreaux J, et al. The level of TACI gene expression in myeloma cells is associated with a signature of microenvironment dependence versus a plasmablastic signature. Blood. 2005;106:1021–1030. doi: 10.1182/blood-2004-11-4512.
- Lee L, et al. Evaluation of B cell maturation antigen as a target for antibody drug conjugate mediated cytotoxicity in multiple myeloma. Br. J. Haematol. 2016;174:911–922. doi: 10.1111/bjh.14145.
- Hsi ED, et al. CS1, a potential new therapeutic antibody target for the treatment of multiple myeloma. Clin. Cancer Res. 2008;14:2775–2784. doi: 10.1158/1078-0432.CCR-07-4246.
- Tai Y-T, et al. Anti-CS1 humanized monoclonal antibody HuLuc63 inhibits myeloma cell adhesion and induces aritibody-dependent cellular cytotoxicity in the bone marrow mitieu. Blood. 2008;112:1329–1337. doi: 10.1182/blood-2007-08-107292.
- Wang X, et al. Lenalidomide enhances the function of CS1 chimeric antigen receptor-redirected T cells against multiple myeloma. Clin. Cancer Res. 2018;24:106–119. doi: 10.1158/1078-0432.CCR-17-0344.
- Hudecek M, et al. Receptor affinity and extracellular domain modifications affect tumor recognition by ROR1-specific chimeric antigen receptor T cells. Clin. Cancer Res. 2013;19:3153–3164. doi: 10.1158/1078-0432.CCR-13-0330.
- Hudecek M, et al. The nonsignaling extracellular spacer domain of chimeric antigen receptors is decisive for in vivo antitumor activity. Cancer Immunol. Res. 2015;3:125–135. doi: 10.1158/2326-6066.CIR-14-0127.
- James JR, Vale RD. Biophysical mechanism of T-cell receptor triggering in a reconstituted system. Nature. 2012;487:64–69. doi: 10.1038/nature11220.
- Williams, M., Tso, Y., Landolfi, N. F., Powers, D. B. & Liu, G. Therapeutic use of anti-CS1 antibodies US (US Patents, 2010). .
- Chen KH, et al. A compound chimeric antigen receptor strategy for targeting multiple myeloma. Leukemia. 2018;32:402–412. doi: 10.1038/leu.2017.302.
- Bouchon A, Cella M, Grierson HL, Cohen JI, Colonna M. Cutting edge: activation of NK cell-mediated cytotoxicity by a SAP-independent receptor of the CD2 family. J. Immunol. 2001;167:5517–5521. doi: 10.4049/jimmunol.167.10.5517.
- Turtle CJ, et al. CD19 CAR-T cells of defined CD4+: CD8+ composition in adult B cell ALL patients. J. Clin. Invest. 2016;1:1–16.
- Berger C, et al. Adoptive transfer of effector CD8+ T cells derived from central memory cells establishes persistent T cell memory in primates. J. Clin. Invest. 2008;118:294–305. doi: 10.1172/JCI32103.
- Sommermeyer D, 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. doi: 10.1038/leu.2015.247.
- Xu Y, et al. Closely-related T-memory stem cells correlate with in-vivo expansion of CAR.CD19-T cells in patients and are preserved by IL-7 and IL-15. Blood. 2014;123:3750–3759. doi: 10.1182/blood-2014-01-552174.
- Frigault MJ, et al. Identification of chimeric antigen receptors that mediate constitutive or inducible proliferation of T cells. Cancer Immunol. Res. 2015;3:356–367. doi: 10.1158/2326-6066.CIR-14-0186.
- Eyquem J, et al. Targeting a CAR to the TRAC locus with CRISPR/Cas9 enhances tumour rejection. Nature. 2017;543:113–117. doi: 10.1038/nature21405.
- Verma V, et al. PD-1 blockade in subprimed CD8 cells induces dysfunctional PD-1+CD38hi cells and anti-PD-1 resistance. Nat. Immunol. 2019;20:1231–1243. doi: 10.1038/s41590-019-0441-y.
- Zelba H, et al. Accurate quantification of T-cells expressing PD-1 in patients on anti-PD-1 immunotherapy. Cancer Immunol. Immunother. 2018;67:1845–1851. doi: 10.1007/s00262-018-2244-7.
- Rosenzweig M, et al. Preclinical data support leveraging CS1 chimeric antigen receptor T-cell therapy for systemic light chain amyloidosis. Cytotherapy. 2017;19:861–866. doi: 10.1016/j.jcyt.2017.03.077.
- Lonial S, et al. Elotuzumab therapy for relapsed or refractory multiple myeloma. N. Engl. J. Med. 2015;373:621–631. doi: 10.1056/NEJMoa1505654.
- Magen H, Muchtar E. Elotuzumab: the first approved monoclonal antibody for multiple myeloma treatment. Ther. Adv. Hematol. 2016;7:187–195. doi: 10.1177/2040620716652862.
- Zonder JA, et al. A phase 1, multicenter, open-label, dose escalation study of elotuzumab in patients with advanced multiple myeloma. Blood. 2013;120:552–559. doi: 10.1182/blood-2011-06-360552.
- Kumar M, Keller B, Makalou N, Sutton RE. Systematic determination of the packaging limit of lentiviral vectors. Hum. Gene Ther. 2001;12:1893–1905. doi: 10.1089/104303401753153947.
- Bos TJ, De Bruyne E, Van Lint S, Heirman C, Vanderkerken K. Large double copy vectors are functional but show a size-dependent decline in transduction efficiency. J. Biotechnol. 2010;150:37–40. doi: 10.1016/j.jbiotec.2010.07.010.
- Qin H, Haso W, Nguyen SM, Fry TJ. Preclinical development of bispecific chimeric antigen receptor targeting both CD19 and CD22. Blood. 2015;126:4427. doi: 10.1182/blood.V126.23.4427.4427.
- Gibson DG, et al. Enzymatic assembly of DNA molecules up to several hundred kilobases. Nat. Methods. 2009;6:343–345. doi: 10.1038/nmeth.1318.
- Brogdon, J. et al. WO2016014565_Treatment of cancer using humanized anti-BCMA CAR.pdf. (US Patents, 2016). .
- Carpenter RO, et al. B cell maturation antigen is a promising target for adoptive T-cell therapy of multiple myeloma. Clin. Cancer Res. 2013;19:2048–2060. doi: 10.1158/1078-0432.CCR-12-2422.
- Oden F, et al. Potent anti-tumor response by targeting. Mol. Oncol. 2015;9:1348–1358. doi: 10.1016/j.molonc.2015.03.010.
- Lee LSH, et al. An APRIL based chimeric antigen receptor to simultaneously target BCMA and TACI in multiple myeloma (MM) has potent activity in vitro and in vivo. Blood. 2016;128:379. doi: 10.1182/blood.V128.22.379.379.
- Chu J, et al. Genetic modification of T cells redirected toward CS1 enhances eradication of myeloma cells. Clin. Cancer Res. 2014;20:3989–4000. doi: 10.1158/1078-0432.CCR-13-2510.
- Yu, J., Hofmeister, C. & Chu, J. WO2014179759_CS1 CAR.pdf. (US Patents, 2014). .
- Wang X, et al. A transgene-encoded cell surface polypeptide for selection, in vivo tracking, and ablation of engineered cells. Blood. 2011;118:1255–1263. doi: 10.1182/blood-2011-02-337360.
Source: PubMed