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Luspatercept (RAP-536) modulates oxidative stress without affecting mutation burden in myelodysplastic syndromes

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Abstract

In low-risk myelodysplastic syndrome (LR-MDS), erythropoietin (EPO) is widely used for the treatment of chronic anemia. However, initial response to EPO has time-limited effects. Luspatercept reduces red blood cell transfusion dependence in LR-MDS patients. Here, we investigated the molecular action of luspatercept (RAP-536) in an in vitro model of erythroid differentiation of MDS, and also in a in vivo PDX murine model with primary samples of MDS patients carrying or not SF3B1 mutation. In our in vitro model, RAP-536 promotes erythroid proliferation by increasing the number of cycling cells without any impact on apoptosis rates. RAP-536 promoted late erythroid precursor maturation while decreasing intracellular reactive oxygen species level. RNA sequencing of erythroid progenitors obtained under RAP-536 treatment showed an enrichment of genes implicated in positive regulation of response to oxidative stress and erythroid differentiation. In our PDX model, RAP-536 induces a higher hemoglobin level. RAP-536 did not modify variant allele frequencies in vitro and did not have any effect against leukemic burden in our PDX model. These results suggest that RAP-536 promotes in vivo and in vitro erythroid cell differentiation by decreasing ROS level without any remarkable impact on iron homeostasis and on mutated allele burden.

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Acknowledgements

Flow cytometry data were obtained on the “Plateforme de cytométrie” of the CHU Grenoble Alpes at the Institute of Biology and Pathology. We thank the zootechnicians of the PHTA facility for animal housing and care.

Funding

We have received a grant from Celgene for this work.

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Author notes

  1. Meunier Mathieu and Chloé Friedrich contributed equally.

Authors and Affiliations

  1. Department of Hematology, CHU Grenoble Alpes, CS10217, 38043, Grenoble cedex 09, France

    Meunier Mathieu & Sophie Park

  2. CNRS UMR 5309, INSERM, U1209, Université Grenoble Alpes, Institute for Advanced Bioscience, 38700, Grenoble, France

    Meunier Mathieu, Johanna Zannoni, Nelly Jerraya, Sophie Rousseaux, Florent Chuffart & Sophie Park

  3. Institut Cochin, Department Development, Reproduction and Cancer, 75014, Paris, France

    Chloé Friedrich & Olivier Kosmider

  4. Université de Paris, INSERM, CNRS, Centre de Recherche Sur L‘Inflammation (CRI), 75018, Paris, France

    Nicolas Ducrot

  5. Laboratoire de Génétique Des Hémopathies, CHU Grenoble Alpes, Grenoble, France

    Tondeur Sylvie

  6. Hematology Department, Hôpital Cochin, Assistance Publique-Hôpitaux de Paris, Université de Paris (APHP-CUP), 75014, Paris, France

    Olivier Kosmider

  7. Université de Toulouse, INSERM, CNRS, Institut Toulousain Des Maladies Infectieuses Et Inflammatoires (Infinity), Université Paul Sabatier (UPS), Toulouse, France

    Zoubida Karim

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  1. Meunier Mathieu
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Contributions

MM and SP wrote the paper. SP supervised the study design. MM, NJ, and JZ performed the in vitro testing. MM engineered the PDX mice model. ND and ZK contributed to iron metabolism measurements in the PDX model. CF and OK performed the molecular testing in the PDX model. ST did the NGS in the in vitro model. FC and SR analyzed the RNA sequencing data.

Corresponding authors

Correspondence to Meunier Mathieu or Sophie Park.

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Ethical approval

All bone marrow samples were collected after written consent in a protocol approval by the institutional ethical review board. Biological samples were stored at CHUGA Biological Resource Center (accreditation number AC 2014–2094, BRIF BB-0033–00069). All applicable international, national, and institutional guidelines for the care and use of animals were followed. The study was approved by our local animals ethical committee and by the French “Ministère de l’enseignement supérieur, de la recherche et de l’innovation.”

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Mathieu, M., Friedrich, C., Ducrot, N. et al. Luspatercept (RAP-536) modulates oxidative stress without affecting mutation burden in myelodysplastic syndromes. Ann Hematol 101, 2633–2643 (2022). https://doi.org/10.1007/s00277-022-04993-7

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