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STEM CELL TRANSPLANTATION

Enhanced thrombin/PAR1 activity promotes G-CSF- and AMD3100-induced mobilization of hematopoietic stem and progenitor cells via NO upregulation

Leukemia volume 35, pages 3334–3338 (2021)Cite this article

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Hematopoietic stem and progenitor cell (HSPC) transplantation is a curative treatment for patients with hematological and malignant diseases. Mobilized peripheral blood (PB) HSPCs are the most widely used source for clinical transplantations. Recent studies, including ours, revealed that traditionally viewed coagulation-related pathways such as thrombin, protease-activated-receptor 1 (PAR1), and Endothelial-Protein C Receptor (EPCR) play essential roles in controlling HSPC-retention-trafficking switch [1,2,3]. Intriguing observations of human PB led us to depict PAR1 expression by circulating mononuclear cells as a marker predicting the efficiency of G-CSF-induced HSPC mobilization and blood-count recovery kinetics of transplanted patients [4]. Understanding molecular mechanisms driven by coagulation pathways to regulate clinical HSPC mobilization is still missing. Herein, we report that the thrombin/PAR1/nitric oxide (NO) axis is a crucial regulatory pathway mediating G-CSF- and AMD3100-induced HSPC mobilization.

To further assess the involvement of thrombin/PAR1 signaling as an essential mechanism mediating HSPC mobilization, we injected mice with another durable mobilizing agent, AMD3100 (Plerixafor), clinically used to induce rapid HSPC mobilization. Similar to G-CSF, AMD3100 increased thrombin activity in the BM shortly after treatment (Supplementary Fig. 1b), and co-treatment with the PAR1-antagonist abrogated HSPC mobilization (Supplementary Fig. 1c). Furthermore, antagonizing PAR1 activity along with AMD3100 treatment significantly reduced the long-term BM repopulation by mobilized HSPCs in a functional, competitive BM transplantation (Supplementary Fig. 1d). Supporting the role played by NO in HSPC mobilization, we detected a decreased NO production in BM-HSPCs upon co-treatment of AMD3100 with PAR1-antagonist (Supplementary Fig. 1e), and limiting NO production by arginine-free-diet attenuated AMD3100-induced HSPC mobilization (Supplementary Fig. 1f).

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Fig. 1: HSPC mobilization involves thrombin/PAR1/NO axis activity.
Fig. 2: Thrombin inhibition reduces EPCR shedding, facilitating HSPC homing and engraftment.

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Acknowledgements

This work was partially supported by the Israel Science Foundation (ISF), a joint grant from the Israel Science Foundation (ISF) and the National Natural Science Foundation of China (NSFC), Minerva Foundation and the Federal German Ministry for Education and Research, Steven B. Rubenstein Research Fund, Weizmann Stem Cell Institute, The WIS Moross Integrated Cancer Center, Asher Pertman and Wayne Pertman, and the Estate of David Levinson.

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  1. These authors contributed equally: Neta Nevo, Lizeth-Alejandra Ordonez-Moreno

Authors and Affiliations

  1. Department of Immunology, Weizmann Institute of Science, Rehovot, Israel

    Neta Nevo, Lizeth-Alejandra Ordonez-Moreno, Shiri Gur-Cohen, Francesca Avemaria, Suditi Bhattacharya, Eman Khatib-Massalha, Mayla Bertagna, Montaser Haddad, Priyasmita Chakrabarti, Tsvee Lapidot & Orit Kollet

  2. Department of Immunology and Microbial Science, Scripps Research, La Jolla, CA, USA

    Wolfram Ruf

  3. Center for Thrombosis and Hemostasis, Johannes Gutenberg-University Medical Center, Mainz, Germany

    Wolfram Ruf

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  1. Neta Nevo
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Contributions

NN and OK reviewed literature, designed and conducted experiments, analyzed results, and wrote the paper. SGC, LAOM, FA, MB, EKM, MH, PC, and SB designed and performed experiments, analyzed data, and reviewed the manuscript. WR participated in designing experiments and reviewed the manuscript. TL reviewed literature, designed the study concept, guided the research, and wrote the paper.

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Correspondence to Tsvee Lapidot or Orit Kollet.

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Nevo, N., Ordonez-Moreno, LA., Gur-Cohen, S. et al. Enhanced thrombin/PAR1 activity promotes G-CSF- and AMD3100-induced mobilization of hematopoietic stem and progenitor cells via NO upregulation. Leukemia 35, 3334–3338 (2021). https://doi.org/10.1038/s41375-021-01194-5

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