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Phenformin increases early hematopoietic progenitors in the Jak2V617F murine model

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Summary

Background. Myeloproliferative neoplasms (MPN) are disorders characterized by an alteration at the hematopoietic stem cell (HSC) level, where the JAK2 mutation is the most common genetic alteration found in classic MPN (polycythemia vera, essential thrombocythemia, and primary myelofibrosis). We and others previously demonstrated that metformin reduced splenomegaly and platelets counts in peripheral blood in JAK2V617F pre-clinical MPN models, which highlighted the antineoplastic potential of biguanides for MPN treatment. Phenformin is a biguanide that has been used to treat diabetes, but was withdrawn due to its potential to cause lactic acidosis in patients. Aims. We herein aimed to investigate the effects of phenformin in MPN disease burden and stem cell function in Jak2V617F-knockin MPN mice. Results. In vitro phenformin treatment reduced cell viability and increased apoptosis in SET2 JAK2V67F cells. Long-term treatment with 40 mg/kg phenformin in Jak2V617F knockin mice increased the frequency of LSK, myeloid progenitors (MP), and multipotent progenitors (MPP) in the bone marrow. Phenformin treatment did not affect peripheral blood counts, spleen weight, megakaryocyte count, erythroid precursors frequency, or ex vivo clonogenic capacity. Ex vivo treatment of bone marrow cells from Jak2V617F knockin mice with phenformin did not affect hematologic parameters or engraftment in recipient mice. Conclusions. Phenformin increased the percentages of LSK, MP, and MPP populations, but did not reduce disease burden in Jak2V617F-knockin mice. Additional studies are necessary to further understand the effects of phenformin on early hematopoietic progenitors.

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Data availability

The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.

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Acknowledgements

The authors would like to thank Dr Nicola Coran for English revision.

Funding

Funding for this work was supported in part by Fundação de Amparo à Pesquisa do Estado de São Paulo/São Paulo Research Foundation (FAPESP Grant #2013/08135–2), Conselho Nacional de Desenvolvimento Científico e Tecnológico/National Counsel of Technological and Scientific Development (CNPq; 2014/50947–7 [INCTC]; and 426633/2018–0), and Coordenação de Aperfeiçoamento de Pessoal de Nível Superior—Brasil (CAPES)—Finance Code 001.

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Authors and Affiliations

  1. Department of Medical Imaging, Hematology, and Oncology, Ribeirão Preto Medical School, University of Sao Paulo, Ribeirão Preto, Brazil

    Antônio Bruno Alves-Silva, Bruna Alves Fenerich, Natasha Peixoto Fonseca, Jaqueline Cristina Fernandes, Juan Luiz Coelho-Silva, Thiago Mantello Bianco, Priscila Santos Scheucher, Lorena Lôbo Figueiredo-Pontes & Fabiola Traina

  2. Center for Cell Based Therapy, São Paulo Research Foundation, Ribeirão Preto, Brazil

    Antônio Bruno Alves-Silva, Bruna Alves Fenerich, Natasha Peixoto Fonseca, Jaqueline Cristina Fernandes, Juan Luiz Coelho-Silva, Diego Antonio Pereira-Martins, Thiago Mantello Bianco, Eduardo Magalhães Rego, Lorena Lôbo Figueiredo-Pontes & Fabiola Traina

  3. Department of Internal Medicine, Ribeirão Preto Medical School, University of Sao Paulo, Ribeirão Preto, Brazil

    Diego Antonio Pereira-Martins & Eduardo Magalhães Rego

  4. Hematology Division, Medical School of University of São Paulo, LIM31, São Paulo, Brazil

    Diego Antonio Pereira-Martins & Eduardo Magalhães Rego

  5. Departament of Pathology and Legal Medicine, University of São Paulo at Ribeirão Preto Medical School, Ribeirão Preto, Brazil

    Fernando Chahud

  6. Department of Pharmacology, Institute of Biomedical Sciences, University of São Paulo, São Paulo, Brazil

    João Agostinho Machado-Neto

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  1. Antônio Bruno Alves-Silva
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Contributions

A.B.A-S designed, executed and analyzed the experiments and prepared the manuscript. B.A.F., N.P.F. participated in experiments and prepared the manuscript. J.C.F provided inputs and participated in experiments. J.L.C-S participated and designed experiments. D.A.P-M., T.M.B., P.S.S. participated in flow cytometry experiments and data analysis. E.M.R. and L.L.F-P provided inputs on experimental designed and interpretation of the data. F.C. participated in histopathology experiments. J.A.M–N., designed, analyzed, provided inputs and prepared the manuscript. F.T supervised and participated in overall design of study, experiments and analyses. All authors reviewed and edited the manuscript.

Corresponding author

Correspondence to Fabiola Traina.

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Ethics approval

All procedures performed in studies involving animals were approved and were following recommendations from Ethical Committee for Animal Research from University of São Paulo at Ribeirão Preto Medical School (183/2018) and followed by Grimace scale for assessing pain in mice.

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Research involving Human Participants and/or Animals

This study did not involved human participants. Studies involving animals were approved by Ethical Committee for Animal Research from University of São Paulo at Ribeirão Preto Medical School (183/2018).

Conflict of interests

Antônio Bruno Alves-Silva declares that he has no competing interests. Bruna Alves Fenerich declares that she has no competing interests. Natasha Peixoto Fonseca declares that she has no competing interests. Jaqueline Cristina Fernandes declares that she has no competing interests. Juan Luiz Coelho-Silva declares that he has no competing interests. Diego Antonio Pereira-Martins declares that he has no competing interests. Thiago Mantello Bianco declares that he has no competing interests. Priscila Santos Scheucher declares that she has no competing interests. Eduardo Magalhães Rego declares that he has no competing interests. Fernando Chahud declares that he has no competing interests. João Agostinho Machado-Neto declares that he has no competing interests. Lorena Lôbo Figueiredo-Pontes declares that she has no competing interests. Fabiola Traina declares that she has no competing interests.

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Alves-Silva, A.B., Fenerich, B.A., Fonseca, N.P. et al. Phenformin increases early hematopoietic progenitors in the Jak2V617F murine model. Invest New Drugs 40, 576–585 (2022). https://doi.org/10.1007/s10637-022-01212-y

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