Circumventing CD19 antigen loss
Chimeric antigen receptor (CAR) T cell treatment for B cell malignancies was pioneered with CD19-targeted CAR T cells. Despite robust clinical responses, relapse due to CD19 antigen loss is common. Qin et al. examined B cell activating factor receptor (BAFF-R) as an alternate CAR T cell target. BAFF-R–targeted CAR T cells could kill multiple human lymphoma and leukemia cell lines, either in vitro or in mice, as well as patient-derived samples. The BAFF-R-CAR T cells could also eradicate tumors lacking CD19. Their compelling preclinical results support the clinical development of BAFF-R–targeted CAR T cells for combating B cell malignancies.
CAR T cells targeting CD19 provide promising options for treatment of B cell malignancies. However, tumor relapse from antigen loss can limit efficacy. We developed humanized, second-generation CAR T cells against another B cell–specific marker, B cell activating factor receptor (BAFF-R), which demonstrated cytotoxicity against human lymphoma and acute lymphoblastic leukemia (ALL) lines. Adoptively transferred BAFF-R-CAR T cells eradicated 10-day preestablished tumor xenografts after a single treatment and retained efficacy against xenografts deficient in CD19 expression, including CD19-negative variants within a background of CD19-positive lymphoma cells. Four relapsed, primary ALLs with CD19 antigen loss obtained after CD19-directed therapy retained BAFF-R expression and activated BAFF-R-CAR, but not CD19-CAR, T cells. BAFF-R-CAR, but not CD19-CAR, T cells also demonstrated antitumor effects against an additional CD19 antigen loss primary patient–derived xenograft (PDX) in vivo. BAFF-R is amenable to CAR T cell therapy, and its targeting may prevent emergence of CD19 antigen loss variants.
Fig. S1. Cytokine release assay.
Fig. S2. BAFF-R-CAR T cell in vitro cytotoxic T lymphocyte assay.
Fig. S3. Preliminary BAFF-R-CAR T cell assessment in vivo.
Fig. S4. CAR T cells validated for CAR expression and cytotoxic T lymphocyte activity.
Fig. S5. CD19-KO clone selection.
Data file S1. Primary data.
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Received: 7 February 2019
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Research reported in this publication included work performed in the Analytical Cytometry Core, Hematopoietic Tissue Biorepository Core, Integrative Genomics Core, and Small Animal Imaging Core supported by the National Cancer Institute of the NIH under award number P30CA033572. The content is solely the responsibility of the authors and does not necessarily represent the official views of the NIH. We thank the Sandra and Edward Meyer Cancer Center PDTX Shared Resource for their support. Funding: We are grateful for the support from the Toni Stephenson Lymphoma Center at Beckman Research Institute of City of Hope. The study was also supported by the Leukemia and Lymphoma Society (LLS): Translational Research Program (TRP 6540-18; PI: L.W.K.), Mantle Cell Lymphoma Research Initiative (MCL 7000-18; PI: L.W.K.), and SCOR grants (SCOR 7011-16 and 7012-16; PI: G.G.I.); the NIH/NCI (SPORE 2P50CA107399; PIs: S.J.F. and L.W.K.; 1R21CA223141; PI: H.Q.); the Department of Defense (CA170783; PI: L.W.K.); and the Hope Portfolio Fund at City of Hope (PI: H.Q.). Research in the Müschen laboratory is funded by the NIH/NCI through the Outstanding Investigator Award R35CA197628 (to M.M.), U10CA180827 (to M.M.), R01CA137060, R01CA157644, R01CA172558, and R01CA213138 (to M.M.); the Howard Hughes Medical Institute HHMI-55108547 (to M.M.); and the Falk Trust through a Falk Medical Research Transformational Award (to M.M.). M.M. is a Howard Hughes Medical Institute (HHMI) Faculty Scholar. Author contributions: H.Q. designed the project, oversaw the experiments, analyzed the data, and wrote the manuscript. Z.D., F.W., W.X., and H.S. conducted the in vitro and in vivo experiments. X.W. and M.W. developed and conducted the CAR T cell degranulation and activation assays. W.A.C. and D.L.S. contributed to the data analysis and manuscript preparation. G.W. and X.S. developed the BAFF-R scFv. F.F. and J.X. developed and conducted the surface antigen density assays. T.I.P., C.-W.C., C.K., L.S., and G.G.I. developed the PDX models. J.Y.S. and I.A. provided the primary tumor samples. M.M., S.J.F., and L.W.K. oversaw the project including the data analysis and manuscript writing. Competing interests: L.W.K. and H.Q. are inventors on patents WO2017214167 and WO 2017214170, submitted by the City of Hope, which is related to this work. L.W.K.: InnoLifes, consultancy and equity ownership; Pepromene Bio, consultancy and equity ownership. H.Q.: InnoLifes, consultancy and equity ownership; Pepromene Bio, consultancy and equity ownership. The other authors declare that they do not have competing interests. Data and materials availability: All data associated with this study are present in the paper or Supplementary Materials. Materials produced in this study are protected by the City of Hope intellectual property patents but will remain available to qualified investigators at other research organizations by establishing a material transfer agreement and in accordance with the NIH principles and guidelines. The following reagents are available from Larry Kwak under a material transfer agreement with the Beckman Research Institute of the City of Hope, subject to third party rights. Reagents: BAFF-R CAR T cells, BAFF-R CAR vector plasmid, JeKo-1-Luciferase, Raji-Luciferase, Z-138-Luciferase, Z-138-Luciferase-CD19KO, MEC-1-CD19KO, Nalm6-Luciferase, and Nalm6-Luciferase-CD19KO.
National Cancer Institute: 2P50CA107399
National Cancer Institute: 1R21CA223141
Leukemia and Lymphoma Society: TRP 6540-18
Leukemia and Lymphoma Society: MCL 7000-18
Leukemia and Lymphoma Society: SCOR 7011-16
Leukemia and Lymphoma Society: SCOR 7012-16
Beckman Research Institute, City of Hope: Hope Portfolio Fund
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