Oncogenic Calreticulin Induces Immune Escape by Stimulating TGFβ Expression and Regulatory T-cell Expansion in the Bone Marrow Microenvironment

Dominik Schmidt, Cornelia Endres, Rouven Hoefflin, Geoffroy Andrieux, Melissa Zwick, Nikolaos Karantzelis, Hans F. Staehle, Janaki Manoja Vinnakota, Sandra Duquesne, Miriam Mozaffari Jovein, Dietmar Pfeifer, Heiko Becker, Bruce R. Blazar, Alexander Zähringer, Justus Duyster, Tilman Brummer, Melanie Boerries, Julian Baumeister, Khalid Shoumariyeh, Juan LiAnthony R. Green, Florian H. Heidel, Itay Tirosh, Heike L. Pahl, Nils Leimkühler, Natalie Köhler, Marcelo A.S. de Toledo, Steffen Koschmieder, Robert Zeiser*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

Abstract

Increasing evidence supports the interplay between oncogenic mutations and immune escape mechanisms. Strategies to counteract the immune escape mediated by oncogenic signaling could provide improved therapeutic options for patients with various malignancies. As mutant calreticulin (CALR) is a common driver of myeloproliferative neoplasms (MPN), we analyzed the impact of oncogenic CALRdel52 on the bone marrow (BM) microenvironment in MPN. Single-cell RNA sequencing revealed that CALRdel52 led to the expansion of TGFβ1-producing erythroid progenitor cells and promoted the expansion of FoxP3+ regulatory T cells (Treg) in a murine MPN model. Treatment with an anti-TGFβ antibody improved mouse survival and increased the glycolytic activity in CD4+ and CD8+ T cells in vivo, whereas T-cell depletion abrogated the protective effects conferred by neutralizing TGFβ. TGFβ1 reduced perforin and TNFα production by T cells in vitro. TGFβ1 production by CALRdel52 cells was dependent on JAK1/2, PI3K, and ERK activity, which activated the transcription factor Sp1 to induce TGFβ1 expression. In four independent patient cohorts, TGFβ1 expression was increased in the BM of patients with MPN compared with healthy individuals, and the BM of patients with MPN contained a higher frequency of Treg compared with healthy individuals. Together, this study identified an ERK/Sp1/TGFβ1 axis in CALRdel52 MPNs as a mechanism of immunosuppression that can be targeted to elicit T-cell-mediated cytotoxicity.

Original languageEnglish
Pages (from-to)2985-3003
Number of pages19
JournalCancer Research
Volume84
Issue number18
DOIs
Publication statusPublished - 15 Sept 2024

Funding

We thank Eileen Barleon for performing immunohistochemistry, Klaus Geiger and Dieter Herchenbach for cell sorting, and Heide Dierbach and Katja Graewe for tissue processing and staining. Miguel Waterhouse for providing healthy control samples. We thank Ann Mullally for kindly providing the mut-Calr and MPN plasmids. The authors greatly acknowledge the team of the Genomics and Proteomics Core Facility, German Cancer Research Center/DKFZ, Heidelberg, Germany, for their sequencing service. N. Karantzelis was supported by a grant from the IMM-PACT-Programme for Clinician Scientists, Department of Medicine II, Medical Center – University of Freiburg and Faculty of Medicine, University of Freiburg, funded by the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) – 413517907. This study was supported by the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) – SFB-1479 – Project ID: 441891347 (to T. Brummer, M. Boerries, N. Köhler, R. Zeiser), DFG Research Unit RU5659 (TARGET MPN; to H.L. Pahl, F.H. Heidel, S. Koschmieder, R. Zeiser), SFB TRR167 (to R. Zeiser and M. Boerries), SFB1160 (to R. Zeiser and M. Boerries), CRC1453 – Project ID 431984000 (M. Boerries), TRR359 – Project ID 491676693 (M. Boerries), FOR 5476 UcarE – Project ID 493802833 (M. Boerries), DFG individual grant 872/4-1 to R. Zeiser, the European Union: EU Proposal n° ERC-2022-ADG Project: 101094168 – AlloCure (ERC Advanced grant to R. Zeiser), ERA-NET Transcan – PIXEL (to R. Zeiser), ERA-NET Transcan – SmartCART (to R. Zeiser), the Deutsche Krebshilfe (grant number 70113473) and the Jose-Carreras Leukemia Foundation grant number DJCLS 09R/2022 (to R. Zeiser), by the DFG under Germany’s Excellence Strategy (CIBSS – EXC-2189 – Project ID 390939984 to N. Köhler and R. Zeiser), INTERREG V European regional development fund (European Union) program (project 3.2 TRIDIAG to R. Zeiser). M. Boerries is supported by the German Federal Ministry of Education and Research (BMBF) within the framework of the Medical Informatics Funding Scheme (PM4Onco FKZ 01ZZ2322A to M. Boerries and EkoEstMed–FKZ 01ZZ2015 to G. Andrieux). BR Blazar was supported in part by NIH grants R01 11879, R37 AI34495, P01 065493 and P01 HL15805. Publisher Copyright: ©2024 American Association for Cancer Research.

All Science Journal Classification (ASJC) codes

  • Oncology
  • Cancer Research

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