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Venetoclax Synergistically Enhances the Anti-leukemic Activity of Vosaroxin Against Acute Myeloid Leukemia Cells Ex Vivo

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Abstract

Background

The survival rate for acute myeloid leukemia remains unacceptably low, in large part owing to resistance to chemotherapy and high rates of relapse. There is an urgent need to develop new therapeutic modalities, in particular such that are tolerated by patients over the age of 60 years, who form the bulk of new acute myeloid leukemia diagnoses. Vosaroxin (SNS-595), a second-generation topoisomerase II inhibitor and DNA intercalating agent, shows promising preclinical and clinical activity against acute myeloid leukemia. Venetoclax (ABT-199), a selective Bcl-2 inhibitor, was recently approved for the treatment of acute myeloid leukemia.

Objective

The objective of this study was to determine the anti-leukemic activity and the underlying molecular mechanisms for the combination of venetoclax and vosaroxin in acute myeloid leukemia cell lines and primary patient samples ex vivo.

Patients and Methods

Using both acute myeloid leukemia cell lines and primary patient samples, annexin V/propidium iodide staining and flow cytometry analyses were used to quantify apoptosis induced by venetoclax or vosaroxin, alone or in combination, with subsequent western blotting analyses to assess levels of Bcl-2 family proteins. Alkaline comet assays were performed to quantify DNA damage induced by the two agents and to determine the effect of venetoclax on DNA repair. Finally, colony-forming assays were conducted on normal human CD34+ cord blood cells and primary acute myeloid leukemia patient samples to determine the effect of venetoclax and vosaroxin on normal hematopoietic and leukemic progenitor cells.

Results

We found that venetoclax and vosaroxin synergistically induced apoptosis in multiple acute myeloid leukemia cell lines. Although vosaroxin could partially abrogate the increase of Mcl-1 protein induced by venetoclax, it could not abrogate the increased binding of Bim to Mcl-1 induced by venetoclax. Cooperative induction of DNA damage occurred within 8 h of treatment with venetoclax plus vosaroxin. Moreover, repair of DNA damage induced by vosaroxin was significantly attenuated by venetoclax. The combination also synergistically induced apoptosis in primary acute myeloid leukemia patient samples and significantly reduced the colony formation capacity of acute myeloid leukemia progenitor cells, while sparing normal hematopoietic progenitor cells.

Conclusions

Vosaroxin and venetoclax synergistically induce apoptosis in acute myeloid leukemia cells and cooperatively target acute myeloid leukemia progenitor cells while sparing normal hematopoietic progenitor cells. Our results support the clinical testing of vosaroxin in combination with venetoclax for treating patients with acute myeloid leukemia, especially in the elderly population.

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Author information

Authors and Affiliations

  1. National Engineering Laboratory for AIDS Vaccine, Key Laboratory for Molecular Enzymology and Engineering, The Ministry of Education, School of Life Sciences, Jilin University, Changchun, People’s Republic of China

    Fangbing Liu, Yongwei Su & Guan Wang

  2. Division of Pediatric Hematology/Oncology, Department of Pediatrics, Children’s Hospital of Michigan, Detroit, MI, USA

    Tristan Knight & Jeffrey W. Taub

  3. Department of Pediatrics, Wayne State University School of Medicine, Detroit, MI, USA

    Tristan Knight, Jeffrey W. Taub & Yubin Ge

  4. Department of Oncology, Wayne State University School of Medicine, 421 E. Canfield, Detroit, MI, 48201, USA

    Holly Edwards & Yubin Ge

  5. Molecular Therapeutics Program, Barbara Ann Karmanos Cancer Institute, Wayne State University School of Medicine, Detroit, MI, USA

    Holly Edwards & Yubin Ge

  6. Department of Pediatric Hematology and Oncology, The First Hospital of Jilin University, Changchun, People’s Republic of China

    Yue Wang

  7. Department of Hematology and Oncology, The First Hospital of Jilin University, Changchun, People’s Republic of China

    Hai Lin

  8. Research Center of Traditional Chinese Medicine, The Affiliated Hospital to Changchun University of Chinese Medicine, 1478 Gongnong Street, Changchun, 130021, Jilin, People’s Republic of China

    Liwei Sun

Authors
  1. Fangbing Liu
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Contributions

YG and LS designed the project and directed the study. FL and YS performed the experiments, data analysis, and interpretation. TK drafted the manuscript. FL, HE, and YG edited the manuscript. HL and YW provided the primary AML patient samples and participated in the coordination of the study. JWT, GW, HE, and YG participated in the data analysis and interpretation. All authors read and approved the final manuscript.

Corresponding authors

Correspondence to Liwei Sun or Yubin Ge.

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Funding

This study was supported by Jilin University, Changchun, China, the Barbara Ann Karmanos Cancer Institute, Wayne State University School of Medicine, and by grants from the National Natural Science Foundation of China (NSFC 31671438 and NSFC 31471295), National Key Research and Development Program of China (2017YFC1702100), Hyundai Hope on Wheels, LaFontaine Family/U Can-Cer Vive Foundation, Kids Without Cancer, Children’s Hospital of Michigan Foundation, Decerchio/Guisewite Family, Justin’s Gift, Elana Fund, Ginopolis/Karmanos Endowment, and the Ring Screw Textron Endowed Chair for Pediatric Cancer Research. The funders had no role in the study design, data collection, analysis and interpretation of data, decision to publish, or preparation of the manuscript.

Conflict of interest

Fangbing Liu, Tristan Knight, Yongwei Su, Holly Edwards, Guan Wang, Yue Wang, Jeffrey W. Taub, Hai Lin, Liwei Sun, and Yubin Ge have no conflicts of interest that are directly relevant to the contents of this article.

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Liu, F., Knight, T., Su, Y. et al. Venetoclax Synergistically Enhances the Anti-leukemic Activity of Vosaroxin Against Acute Myeloid Leukemia Cells Ex Vivo. Targ Oncol 14, 351–364 (2019). https://doi.org/10.1007/s11523-019-00638-4

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