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IMMUNOTHERAPY

CD19/BAFF-R dual-targeted CAR T cells for the treatment of mixed antigen-negative variants of acute lymphoblastic leukemia

Leukemia volume 36, pages 1015–1024 (2022)Cite this article

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Abstract

Chimeric antigen receptor (CAR) T cells targeting CD19 mediate potent antitumor effects in B-cell malignancies including acute lymphoblastic leukemia (ALL), but antigen loss remains the major cause of treatment failure. To mitigate antigen escape and potentially improve the durability of remission, we developed a dual-targeting approach using an optimized, bispecific CAR construct that targets both CD19 and BAFF-R. CD19/BAFF-R dual CAR T cells exhibited antigen-specific cytokine release, degranulation, and cytotoxicity against both CD19−/− and BAFF-R−/− variant human ALL cells in vitro. Immunodeficient mice engrafted with mixed CD19−/− and BAFF-R−/− variant ALL cells and treated with a single dose of CD19/BAFF-R dual CAR T cells experienced complete eradication of both CD19−/− and BAFF-R−/− ALL variants, whereas mice treated with monospecific CD19 or BAFF-R CAR T cells succumbed to outgrowths of CD19−/BAFF-R+ or CD19+/BAFF-R− tumors, respectively. Further, CD19/BAFF-R dual CAR T cells showed prolonged in vivo persistence, raising the possibility that these cells may have the potential to promote durable remissions. Together, our data support clinical translation of BAFF-R/CD19 dual CAR T cells to treat ALL.

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Fig. 1: Development of prototype BAFF-R/CD19 dual-targeting CAR T cells.
Fig. 2: Functional evaluation of CD19/BAFF-R dual-targeting CAR T cells.
Fig. 3: Antitumor activity of CD19-BAFF-R(l) dual-targeting CAR T cells manufactured in a clinically relevant platform.
Fig. 4: Comparison of CD19-BAFF-R(l) dual-targeting CAR with CD19 and BAFF-R single CAR in a mixed tumor model.
Fig. 5: Analysis of residual tumor and CAR T cells in mouse peripheral blood.
Fig. 6: Dual CAR T cells exhibited efficient cytotoxicity against primary ALL cells.

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Acknowledgements

The Small Animal Imaging Core is supported by the National Cancer Institute of the National Institutes of Health (P30CA033572); Toni Stephenson Lymphoma Center (PI: L.W.K.); Leukemia and Lymphoma Society Mantle cell lymphoma Research Initiative (SCOR 7000-18; PI: L.W.K.; Project Leaders: S.J.F.; X.W.; E.L.B.); Department of Defense (CA170783, PI: L.W.K.). Lymphoma SPORE (P50 CA107399-11, PI: S.J.F; L.W.K; Project leaders: X.W.; R.N.).

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  1. These authors contributed equally: Xiuli Wang, Zhenyuan Dong.

Authors and Affiliations

  1. Cellular Immunotherapy Center, Department of Hematology and Hematopoietic Cell Transplantation, Beckman Research Institute of City of Hope, Duarte, CA, 91010, USA

    Xiuli Wang, Dennis Awuah, Wen-Chung Chang, Vibhuti Vyas, Ibrahim Aldoss & Stephen J. Forman

  2. Toni Stephenson Lymphoma Center, Department of Hematology and Hematopoietic Cell Transplantation, Beckman Research Institute of City of Hope, Duarte, CA, 91010, USA

    Zhenyuan Dong, Wesley A. Cheng, Soung-Chul Cha, Aaron J. Anderson, Tiantian Zhang, Zhe Wang, Szymon J. Szymura, Benjamin Z. Kuang, Larry W. Kwak & Hong Qin

  3. Department of Clinical and Translational Project Development, City of Hope, Duarte, CA, 91010, USA

    Mary C. Clark

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Contributions

L.W.K., S.J.F., X.W., I.A., and H.Q. designed and directed the study, analyzed and organized the data, and wrote the manuscript. W.C.C., V.V., D.A. designed the vectors and produced lentivirus. D.A., Z.D., S.C., T.Z., Z.W., S.S., B.K., and A.A. performed experimental work in mice and immune assays. M.C.C. and W.A.C. reviewed and edited the manuscript. All authors read and approved the final version.

Corresponding authors

Correspondence to Stephen J. Forman or Larry W. Kwak.

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Competing interests

Sources of support include the National Cancer Institute of the National Institutes of Health (P30CA033572) supporting the Small Animal Imaging Core, National Cancer Institute Lymphoma SPORE (PI: L.W.K.), Leukemia and Lymphoma Society Mantle cell lymphoma Research Initiative (SCOR 7000-18; PI: L.W.K.; Project Leaders: S.J.F.; X.W.), Department of Defense (CA170783, PI: L.W.K.). X.W. is a paid consultant for Pepromene Bio, Inc.; L.W.K. and H.Q. are paid consultants and have equity in Pepromene Bio, Inc. The other authors report no disclosures.

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Wang, X., Dong, Z., Awuah, D. et al. CD19/BAFF-R dual-targeted CAR T cells for the treatment of mixed antigen-negative variants of acute lymphoblastic leukemia. Leukemia 36, 1015–1024 (2022). https://doi.org/10.1038/s41375-021-01477-x

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