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Deoxycytidine-kinase knockdown as a novel myeloprotective strategy in the context of fludarabine, cytarabine or cladribine therapy

Leukemia volume 29, pages 2266–2269 (2015)Cite this article

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Myeloprotective gene therapy aims at the protection of the lymphohematopoietic system from the toxicity of antineoplastic chemotherapy by overexpression of drug resistance (CTX-R) genes, and a number of such CTX-R genes conferring protection against a variety of anticancer agents have been identified.1 Recently efficient protection from alkylating agent-induced myelotoxicity has been demonstrated following gene transfer of the CTX-R gene mutant O6-methylguanine methyltransferase (mutMGMT) in a cohort of glioblastoma patients treated with temozolomide.2, 3 In this study profound in vivo enrichment of genetically modified hematopoietic cells and clinical benefits in comparison to a disease matched control group was demonstrated establishing a proof-of-concept for the clinical applicability and efficacy of myeloprotective gene therapy approaches.

Other CTX-R genes code for enzymes involved in nucleotide metabolism. Thus, in the pyrimidine nucleotide salvage pathway, deoxycytidine kinase (dCK) catalyzes the phosphorylation of deoxycytidine as well as its cytotoxic analogs cytarabine (Ara-C), fludarabine and cladribine prior to their incorporation into the DNA, while cytidine deaminase (CDD) prevents this phosphorylation cascade. Increased intracellular CDD levels as well as low-level expression of dCK confer resistance to Ara-C in lymphomas and leukemic blast cells4, 5 and overexpression of CDD has been demonstrated to induce high level resistance in hematopoietic cells to Ara-C in vitro as well as in in vivo animal models.6, 7, 8 However, CDD overexpression is not effective in the context of the halogenated, deaminase-resistant pyrimidine analogs fludarabine or cladribine. In contrast, downmodulation of dCK can be expected to confer resistance to all three drugs. To prove this hypothesis, we here have investigated the short interfering RNA-mediated knockdown of dCK in human hematopoietic cells as a first step to an effective myeloprotective strategy also in the context of fludarabine or cladribine therapy.

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Acknowledgements

We thank Matthias Ballmaier and his team from the Core-Facility Cell-Sorting of the Hannover Medical School for cell sorting and Doreen Lüttge, as well as Annette Garbe (the Hannover Medical School) for excellent technical assistance. This work was supported by grants from the Deutsche Forschungsgemeinschaft: Cluster of Excellence REBIRTH (Exc 62/1), SPP1230 grant MO 886/3-1 (TM), grant MO 886/4-1 (TM), the EU framework program grant PERSIST (CHB) and Hannover Biomedical Research School (HBRS; DFG, GSC 108).

Author Contributions

NL, KC, AS and TM designed the study; NL, KC, SB, MH and RP performed laboratory work; VK performed mass spectrometry; NH assisted in cloning and helped to design cloning strategy; HK performed whole plate images; NL, RP, SB, DD and TM wrote the manuscript; all authors critically evaluated the manuscript and approved its content.

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Author notes

  1. R Phaltane

    Present address: 9Current address: Institute of Biological and Medical Imaging, Technische Universität München and Helmholtz Zentrum München, Munich, Germany.,

  2. N Heinz

    Present address: 10Current address: Research Group for Gene Modification in Stem Cells, LOEWE Center for Cell and Gene Therapy Frankfurt, Paul-Ehrlich-Institut, Langen, Germany.,

  3. N Lachmann and K Czarnecki: These authors have contributed equally to this work.

Authors and Affiliations

  1. Reprogramming and Gene Therapy Group, REBIRTH Cluster of Excellence, Hannover Medical School, Hannover, Germany

    N Lachmann, K Czarnecki, S Brennig, R Phaltane, M Heise & T Moritz

  2. Institute of Experimental Hematology, Hannover Medical School, Hannover, Germany

    N Lachmann, K Czarnecki, S Brennig, R Phaltane, M Heise, N Heinz, A Schambach & T Moritz

  3. Department of Cardiothoracic, Leibniz Research Laboratories for Biotechnology and Artificial Organs (LEBAO), Transplantation and Vascular Surgery, Hannover Medical School, Hannover, Germany

    H Kempf

  4. Department of Pediatric Hematology and Oncology, Center for Child and Adolescent Medicine, Rheinische Friedrich-Wilhelms University, Bonn, Germany

    D Dilloo

  5. Institute of Pharmacology, Research Core Unit Metabolomics, Hannover Medical School, Hannover, Germany

    V Kaever

  6. Division of Hematology/Oncology, Boston Children's Hospital, Harvard Medical School, Boston, MA, USA

    A Schambach

  7. Department of Hematology, Hemostasis, Oncology and Stem Cell Transplantation, Hannover Medical School, Hannover, Germany

    M Heuser

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Correspondence to T Moritz.

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Lachmann, N., Czarnecki, K., Brennig, S. et al. Deoxycytidine-kinase knockdown as a novel myeloprotective strategy in the context of fludarabine, cytarabine or cladribine therapy. Leukemia 29, 2266–2269 (2015). https://doi.org/10.1038/leu.2015.108

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