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Ceramide synthase 6 impacts T-cell allogeneic response and graft-versus-host disease through regulating N-RAS/ERK pathway

Leukemia volume 36, pages 1907–1915 (2022)Cite this article

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Abstract

Allogeneic hematopoietic cell transplantation (allo-HCT) is an effective immunotherapy for various hematologic malignancies, predominantly through potent graft-versus-leukemia (GVL) effect. However, the mortality after allo-HCT is because of relapse of primary malignancy and followed by graft-vs-host-disease (GVHD) as a major cause of transplant-related mortality. Hence, strategies to limit GVHD while preserving the GVL effect are highly desirable. Ceramide, which serves a central role in sphingolipid metabolism, is generated by ceramide synthases (CerS1–6). In this study, we found that genetic or pharmacologic targeting of CerS6 prevented and reversed chronic GVHD (cGVHD). Furthermore, specific inhibition of CerS6 with ST1072 significantly ameliorated acute GVHD (aGVHD) while preserving the GVL effect, which differed from FTY720 that attenuated aGVHD but impaired GVL activity. At the cellular level, blockade of CerS6 restrained donor T cells from migrating into GVHD target organs and preferentially reduced activation of donor CD4 T cells. At the molecular level, CerS6 was required for optimal TCR signaling, CD3/PKCθ co-localization, and subsequent N-RAS activation and ERK signaling, especially on CD4+ T cells. The current study provides rationale and means for targeting CerS6 to control GVHD and leukemia relapse, which would enhance the efficacy of allo-HCT as an immunotherapy for hematologic malignancies in the clinic.

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Fig. 1: Absence of CerS6 on donor T cells or inhibition of CerS6 ameliorates cGVHD development.
Fig. 2: ST1072 modulates B- and T-cell functions during cGVHD development.
Fig. 3: The effects of ST1072 or FTY720 on GVH and GVL responses.
Fig. 4: ST1072 treatment modulates T-cell expansion and migration after allo-BMT.
Fig. 5: The role of CerS6 in proximal TCR signaling.
Fig. 6: The effect of CerS6 on CD3 and PKCθ capping in CD4 T cells after TCR stimulation.
Fig. 7: Preventative treatment with ST1072 reduces GVHD in NSG xenograft mouse model.

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Acknowledgements

We would also like to thank the Flow Cytometry Core, Small Animal Imaging Core, Lipidomics Shared Resources and Cell & Molecular Imaging Facilities at Medical University of South Carolina for the assistance. This work is supported in part by SC SmartState Cancer Stem Cell Biology & Therapy Program and by R01 grants from the National Institutes of Health including AI118305 and HL140953 (X-ZY); CA214641, DE016572, P01 CA203628, and SC SmartState Endowment in Lipidomics and Drug Discovery (BO); and the DFG GRK2158 (HS).

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  1. These authors contributed equally: M. Hanief Sofi, Linlu Tian.

Authors and Affiliations

  1. Department of Microbiology and Immunology, Medical University of South Carolina, Charleston, SC, USA

    M. Hanief Sofi, Linlu Tian, Steven Schutt, Hee-Jin Choi, Yongxia Wu, David Bastian, Taylor Ticer, Xiaohui Sui & Xue-Zhong Yu

  2. Department of Microbiology and Immunology, Medical College of Wisconsin, Milwaukee, WI, USA

    Linlu Tian, Hee-Jin Choi, Yongxia Wu & Xue-Zhong Yu

  3. Department of Cell and Molecular Pharmacology and Experimental Therapeutics, Medical University of South Carolina, Charleston, SC, USA

    Imran Khan & John P. O’Bryan

  4. Department of Biochemistry and Molecular Biology, Medical University of South Carolina, Charleston, SC, USA

    Mohamed Faisal Kassir, Firdevs Cansu Atilgan, Jisun Kim & Besim Ogretmen

  5. Institute of Pharmaceutical and Medicinal Chemistry, Heinrich Heine University Düsseldorf, Duesseldorf, Germany

    Aleksandra Zivkovic & Holger Stark

  6. Department of Surgery, Medical University of South Carolina, Charleston, SC, USA

    Shikhar Mehrotra

  7. Clinical Research Division, Fred Hutchinson Cancer Research Center, Seattle, WA, USA

    Paul J. Martin

  8. The Cancer Center, Medical College of Wisconsin, Milwaukee, WI, USA

    Xue-Zhong Yu

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Contributions

MHS, LT, and X-ZY participated in designing research studies, MHS, SS, IK, HC, YW, DB, TT, MFK, FCA, KJ, and XS participated in conducting experiments and acquiring data. MHS, LT, and XZY participated in analyzing and interpreting data and wrote the manuscript. AZ and HS provided ST1072 inhibitor. PJM provided deidentified patient plasma samples and related information and participated in clinical data analysis and interpretation. SM, JO, and BO participated in interpreting data and editing the manuscript.

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Correspondence to Xue-Zhong Yu.

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Sofi, M.H., Tian, L., Schutt, S. et al. Ceramide synthase 6 impacts T-cell allogeneic response and graft-versus-host disease through regulating N-RAS/ERK pathway. Leukemia 36, 1907–1915 (2022). https://doi.org/10.1038/s41375-022-01581-6

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