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Everolimus in combination with Imatinib overcomes resistance in Chronic myeloid leukaemia

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Abstract

Although Imatinib and other tyrosine kinase inhibitors (TKIs) have excellent results, the appearance of resistance is a problem in chronic myeloid leukaemia (CML). PI3K/AKT/mTOR pathway is activated by BCR-ABL playing a crucial rule in CML. This study aimed to evaluate the therapeutic potential of Everolimus, in CML models sensitive and resistant to Imatinib. We used one CML cell line sensitive to Imatinib (K562) and two resistant (K562-RC and K56-RD). Cell lines were treated with Everolimus alone and in combination with Imatinib. Cell viability was analysed by resazurin assay. Cell death and cell cycle were analysed by flow cytometry. Additionally, we also studied peripheral blood samples obtained from 52 patients under TKI treatment. Everolimus reduced cell line viability in sensitive (IC50 = 20 µM) and resistant models (K562-RC, IC50 = 25 µM; K562-RD, IC50 = 30 µM). This drug induced cell death by apoptosis and cell cycle arrest in G0/G1 phase. Everolimus also reduced cell viability by increasing apoptosis of haematopoietic stem cells (CD34+ cells) with low cytotoxicity to lymphocytes. Everolimus at 25 µM increased apoptotic cells 18.7% in CD34+ cells and only 8% in lymphocytes. The response to Everolimus was influenced by TKI treatment, with a better response in samples from patients under 2nd and 3rd generation TKI and with less toxicity to lymphocytes. Our results reveal that Everolimus induce cell death in CML cells sensitive and resistant to Imatinib, with low cytotoxicity to normal cells, suggesting that Everolimus could be an alternative targeted therapeutic approach in CML patients, even in cases of Imatinib resistance.

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Acknowledgements

The present work was supported by CIMAGO—Center of Investigation on Environment, Genetics and Oncobiology, Faculty of Medicine, University of Coimbra, Portugal (Project 18/12), by funds from FEDER through the Operational Program Competitiveness Factors—COMPETE, and by Portuguese funds through FCT—Foundation for Science and Technology—under the strategic projects from FCT/MCTES/PIDDAC (CNC.IBILI, Center Reference: UID/NEU/04539/2013). RA was supported by Portuguese Foundation to Science and Technology (FCT) with a PhD Grant (SFRH/BD/51994/2012).

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Authors and Affiliations

  1. Laboratory of Oncobiology and Hematology and University Clinic of Hematology, Faculty of Medicine, University of Coimbra, FMUC, Azinhaga de Santa Comba-Celas, 3000-548, Coimbra, Portugal

    Raquel Alves, Ana Cristina Gonçalves, Joana Jorge & Ana B. Sarmento-Ribeiro

  2. Coimbra Institute for Clinical and Biomedical Research (iCBR) - Group of Environment Genetics and Oncobiology (CIMAGO), FMUC, Coimbra, Portugal

    Raquel Alves, Ana Cristina Gonçalves, Joana Jorge, José M. Nascimento Costa & Ana B. Sarmento-Ribeiro

  3. Center for Neuroscience and Cell Biology (CNC.IBILI), University of Coimbra, Coimbra, Portugal

    Raquel Alves, Ana Cristina Gonçalves, Joana Jorge & Ana B. Sarmento-Ribeiro

  4. Centre for Functional Ecology (CFE), Department of Life Sciences, University of Coimbra, Coimbra, Portugal

    Joana Alves & António Alves da Silva

  5. Clinical Hematology Department, Centro Hospitalar Universitário de Coimbra (CHUC), Coimbra, Portugal

    Paulo Freitas-Tavares & Ana B. Sarmento-Ribeiro

  6. Hospital da Luz, Lisbon, Portugal

    António M. Almeida

  7. CIIS (Centro de Investigação Interdisciplinar em Saúde, Universidade Católica Portuguesa de Lisboa, Lisbon, Portugal

    António M. Almeida

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  1. Raquel Alves
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Contributions

RA, AA, and ABSR designed the experiments. RA and ACG drafted the manuscript. RA, ACG, and JJ performed the experiments. JA and AAS executed the statistical analyses. PFT recruited and provided the clinical information of the participants. JNC, AA, and ABSR revised the manuscript. All authors read and approved the final manuscript.

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Correspondence to Ana B. Sarmento-Ribeiro.

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Alves, R., Gonçalves, A.C., Jorge, J. et al. Everolimus in combination with Imatinib overcomes resistance in Chronic myeloid leukaemia. Med Oncol 36, 30 (2019). https://doi.org/10.1007/s12032-019-1253-5

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