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Novel inhibitor of hematopoietic cell kinase as a potential therapeutic agent for acute myeloid leukemia

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Abstract

Myelodysplastic syndromes (MDS) and acute myeloid leukemia (AML) are characterized by risk of relapses, poor survival, unwanted side effects and high toxicity with the current therapies. In light of these facts, there are efforts to develop new drugs specific for deregulated molecules that participate in leukemia pathogenesis. Hematopoietic cell kinase (HCK), an Src kinase family member, is overexpressed on hematopoietic stem cells of MDS and de novo AML patients and involved in the oncogenic process. Thus, we investigated in vitro, ex vivo and in vivo effects of a novel chemical compound targeting HCK inhibition (iHCK-37), in combination with the most used drugs for the treatment of MDS and de novo AML, 5-Azacytidine and Cytarabine. Herein, the combination treatment with iHCK-37 and 5-Azacytidine or Cytarabine demonstrated additive effects against leukemia cells, compared to either drug alone. iHCK-37 plus 5-Azacytidine or Cytarabine treatment was able to reduce the activation of oncogenic pathways, MAPK/ERK and PI3K/AKT, leading to reduction of ERK and AKT phosphorylation, and increased BAX and decreased BCL-XL protein expression. Moreover, treatment with iHCK-37 reduced MDS and AML CD34-positive cell numbers inside a 3D-structure but did not affect normal CD34-positive cell numbers. In vivo analysis showed that leukemic mice treated with iHCK-37 had reduced ERK and AKT proteins phosphorylation levels and leukocyte numbers. In conclusion, the iHCK-37 inhibitor has anti-neoplastic activity in leukemia cells without altering apoptosis and survival rate of normal cells, suggesting on-target malignant cell killing activity as a single agent or in combination with 5-Azacytidine or Cytarabine.

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Acknowledgements

The authors would like to thank Raquel S Foglio (Hematology and Transfusion Medicine Center, University of Campinas, São Paulo, Brazil) for the English revision; Ana Leda Longhini (University of Alabama, Birmingham, AL, USA) and Irene Santos (Hematology and Transfusion Medicine Center, University of Campinas, São Paulo, Brazil) for flow cytometry assistance; Renata Giardini Rosa and Juares Ednaldo Romero Bianco for the development of scaffold material; and Alessio Molinari and Maurizzio Bottas (in memoriam) (Dipartimento di Biotecnologie, Chimica e Farmacia-Università degli Studi di Siena, Siena, Italy) for the synthesis of iHCK-37 compound. We also thank the National Institute of Science and Technology on Photonics Applied to Cell Biology (INFABIC) at UNICAMP for the access to equipment and confocal assistance and analysis (FAPESP Grants 2014/50938-8 and CNPq Grants 465699/2014-6) and the Hemostasis Laboratory of Hematology and Transfusion Medicine Center (FAPESP Grants 2016/14172-6) at UNICAMP for cell proliferation analysis by IncuCyte S3 System (Sartorius). The authors also thank CNPq, FAPESP, and INCT-Sangue for the financial support.

Funding

This study was funded by grants from the Fundação de Amparo a Pesquisa do Estado de São Paulo (FAPESP Grants 2011/22376–7, and 2017/21801–2) and Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq).

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Authors and Affiliations

  1. Hematology and Transfusion Medicine Center, University of Campinas/Hemocentro-Unicamp, Universitária Zeferino Vaz - Barão Geraldo, Rua Carlos Chagas, 480 - Cidade Campinas, São Paulo, Brazil

    Fernanda Marconi Roversi, Maura Lima Pereira Bueno, Cristiane Okuda Torello, Rodrigo Nato Shiraishi, Fernando Viera Pericole, Karla Priscila Ferro, Adriana Santos Silva Duarte & Sara Teresinha Olalla Saad

  2. Department of Ophthalmology and Visual Sciences, University of Alabama at Birmingham-UAB, Birmingham, AL, USA

    Juliete Aparecida Francisco da Silva

  3. Department of Pathology, Faculty of Medical Sciences, University of Campinas-Unicamp, Campinas, São Paulo, Brazil

    Guilherme Rossi Assis-Mendonça

  4. Division of Hematology, Hospital das Clínicas da Faculdade de Medicina (HCFMUSP), University of São Paulo Medical School, University of São Paulo, São Paulo, Brazil

    Eduardo Magalhães Rego

Authors
  1. Fernanda Marconi Roversi
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  2. Maura Lima Pereira Bueno
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  3. Juliete Aparecida Francisco da Silva
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  5. Cristiane Okuda Torello
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  6. Rodrigo Nato Shiraishi
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  7. Fernando Viera Pericole
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  8. Karla Priscila Ferro
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  9. Adriana Santos Silva Duarte
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  10. Eduardo Magalhães Rego
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  11. Sara Teresinha Olalla Saad
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Contributions

Fernanda Marconi Roversi was the principal investigator of this study, designing and performing the experiments, collecting, analyzing and interpreting data, and writing the manuscript; Maura Lima Pereira Bueno, Juliete Aparecida Francisco da Silva, Guilherme Rossi Assis-Mendonça contributed in the experiments and in the writing of the manuscript; Cristiane Okuda Torello, Rodrigo Nato Shiraishi, Karla Priscila Ferro, Eduardo Magalhães Rego contributed to mice experiments; Adriana Santos Silva Duarte carried out the cell separation; Fernando Viera Pericole acted as the reference physician and helped to analyze the data; Sara Teresinha Olalla Saad provided the study conception, directed the research, provided financial support, revised and gave final approval of the manuscript. All the authors read and approved the final manuscript and agreed that they are accountable for all the aspects of the work and assure the accuracy and integrity of the work.

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Correspondence to Sara Teresinha Olalla Saad.

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Roversi, F.M., Bueno, M.L.P., da Silva, J.A.F. et al. Novel inhibitor of hematopoietic cell kinase as a potential therapeutic agent for acute myeloid leukemia. Cancer Immunol Immunother 71, 1909–1921 (2022). https://doi.org/10.1007/s00262-021-03111-2

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