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Chronic lymphocytic leukemia

Siglec-6 is a target for chimeric antigen receptor T-cell treatment of chronic lymphocytic leukemia

Leukemia volume 35, pages 2581–2591 (2021)Cite this article

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Abstract

The only current curative treatment for chronic lymphocytic leukemia (CLL) is allogenic hematopoietic stem cell transplantation. Chimeric antigen receptor treatment targeting CD19 for CLL achieved some complete responses, suggesting the need for alternative or combinational therapies to achieve a more robust response. In this work, we evaluated CAR-T cells specific for Siglec-6, an antigen expressed in CLL, as a novel CAR-T cell treatment for CLL. We found that detection of SIGLEC6 mRNA and Siglec-6 protein is highly restricted to placenta and immune cells in other tissues and it is not expressed in hematopoietic stem cells. We generated CAR-T cells specific for Siglec-6 based on the sequence of the fully human anti-Siglec-6 antibody (JML1), which was identified in a CLL patient that was cured after allo-hematopoietic stem cell transplantation (alloHSCT), and observed that it specifically targeted CLL cells in vitro and in a xenograft mouse model. Interestingly, a short hinge region increased the activity of CAR-T cells to target cells expressing higher Siglec-6 levels but similarly targeted CLL cells expressing lower Siglec-6 levels in vitro and in vivo. Our results identify a novel CAR-T cell therapy for CLL and establish Siglec-6 as a possible target for immunotherapy.

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Fig. 1: Siglec-6 expression in normal tissue and cells.
Fig. 2: Siglec-6 Immunohistochemistry in healthy human tissue.
Fig. 3: JML1-CAR constructs recognize Siglec-6.
Fig. 4: JML1-CAR-short-T-cells elicit higher activation than JML1-CAR-T-cells.
Fig. 5: JML1-CAR-short and JML1-CAR T-cells have cytotoxic activity against CLL cells in vitro.
Fig. 6: JML1-CAR-short and JML1-CAR have antitumor activity in a xenograft mouse model of CLL.

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Acknowledgements

We thank Dr. James N. Kochenderfer for providing the CD19-CAR construct. This work was supported by an NIH, Bench to Bedside and back Program (Grant ID: 600897) to DK, REG, SP, and CR. CR also acknowledges funding by NIH grant R21 CA229961.

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Authors and Affiliations

  1. Experimental Transplantation and Immunotherapy Branch, Center for Cancer Research, National Cancer Institute, NIH, Bethesda, MD, 20892, USA

    Damian Kovalovsky, Jeong Heon Yoon, Samantha Simon, Elisaveta Voynova & Ronald E. Gress

  2. Department of Immunology and Microbiology, The Scripps Research Institute, Jupiter, FL, 33458, USA

    Matthew G. Cyr & Christoph Rader

  3. Laboratory of Lymphoid Malignancies, National Heart, Lung and Blood Institute. NIH, Bethesda, MD, 20892, USA

    Adrian Wiestner

  4. Laboratory of Pathology, Center for Cancer Research, National Cancer Institute, NIH, Bethesda, MD, 20892, USA

    Julie Alejo & Stefania Pittaluga

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  1. Damian Kovalovsky
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Contributions

DK supervised, performed experiments, and wrote the manuscript, JHY, SS, EV, MGC performed experiments, AW provided the primary CLL samples and scientific expertise, CR developed monoclonal antibody JML1, supervised experiments, edited the manuscript, and provided scientific expertise, JA and SP performed immunohistochemistry, and REG provided scientific guidance.

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Correspondence to Damian Kovalovsky.

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Conflict of interest

CR is an inventor of U.S. Patent 8,877,199 which claims monoclonal antibody JML1. All other authors declare no competing interests.

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Kovalovsky, D., Yoon, J.H., Cyr, M.G. et al. Siglec-6 is a target for chimeric antigen receptor T-cell treatment of chronic lymphocytic leukemia. Leukemia 35, 2581–2591 (2021). https://doi.org/10.1038/s41375-021-01188-3

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