Differences between intrinsic and acquired nucleoside analogue resistance in acute myeloid leukaemia cells.

Autor: Rothenburger T; Institute for Medical Virology, Goethe-University, Frankfurt am Main, Germany.; Faculty of Biological Sciences, Goethe-University, Frankfurt am Main, Germany., Thomas D; Pharmazentrum frankfurt/ZAFES, Institute of Clinical Pharmacology, Goethe University of Frankfurt, Frankfurt, Germany., Schreiber Y; Pharmazentrum frankfurt/ZAFES, Institute of Clinical Pharmacology, Goethe University of Frankfurt, Frankfurt, Germany., Wratil PR; Max von Pettenkofer Institute & Gene Center, Virology, National Reference Center for Retroviruses, Faculty of Medicine, LMU München, Munich, Germany.; German Center for Infection Research (DZIF), Partner Site Munich, Munich, Germany., Pflantz T; Max von Pettenkofer Institute & Gene Center, Virology, National Reference Center for Retroviruses, Faculty of Medicine, LMU München, Munich, Germany.; German Center for Infection Research (DZIF), Partner Site Munich, Munich, Germany., Knecht K; Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, CT, USA., Digianantonio K; Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, CT, USA., Temple J; Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, CT, USA., Schneider C; Department of Pediatric Oncology, Dana-Farber Cancer Institute, Harvard Medical School, Boston, MA, USA., Baldauf HM; Max von Pettenkofer Institute & Gene Center, Virology, National Reference Center for Retroviruses, Faculty of Medicine, LMU München, Munich, Germany., McLaughlin KM; School of Biosciences, University of Kent, Canterbury, UK., Rothweiler F; Institute for Medical Virology, Goethe-University, Frankfurt am Main, Germany., Bilen B; Faculty of Biological Sciences, Goethe-University, Frankfurt am Main, Germany., Farmand S; Faculty of Biological Sciences, Goethe-University, Frankfurt am Main, Germany., Bojkova D; Institute for Medical Virology, Goethe-University, Frankfurt am Main, Germany., Costa R; Institute for Medical Virology, Goethe-University, Frankfurt am Main, Germany., Ferreirós N; Pharmazentrum frankfurt/ZAFES, Institute of Clinical Pharmacology, Goethe University of Frankfurt, Frankfurt, Germany., Geisslinger G; Pharmazentrum frankfurt/ZAFES, Institute of Clinical Pharmacology, Goethe University of Frankfurt, Frankfurt, Germany.; Fraunhofer Institute for Molecular Biology and Applied Ecology (IME), Project group Translational Medicine and Pharmacology (TMP), Frankfurt am Main, Germany., Oellerich T; Department of Hematology/Oncology, Goethe-University, Frankfurt am Main, Germany.; Molecular Diagnostics Unit, Frankfurt Cancer Institute, Frankfurt am Main, Germany.; German Cancer Consortium/German Cancer Research Center, Heidelberg, Germany., Xiong Y; Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, CT, USA., Keppler OT; Max von Pettenkofer Institute & Gene Center, Virology, National Reference Center for Retroviruses, Faculty of Medicine, LMU München, Munich, Germany.; German Center for Infection Research (DZIF), Partner Site Munich, Munich, Germany., Wass MN; School of Biosciences, University of Kent, Canterbury, UK., Michaelis M; School of Biosciences, University of Kent, Canterbury, UK. M.Michaelis@kent.ac.uk., Cinatl J Jr; Institute for Medical Virology, Goethe-University, Frankfurt am Main, Germany. Cinatl@em.uni-frankfurt.de.
Jazyk: angličtina
Zdroj: Journal of experimental & clinical cancer research : CR [J Exp Clin Cancer Res] 2021 Oct 12; Vol. 40 (1), pp. 317. Date of Electronic Publication: 2021 Oct 12.
DOI: 10.1186/s13046-021-02093-4
Abstrakt: Background: SAMHD1 mediates resistance to anti-cancer nucleoside analogues, including cytarabine, decitabine, and nelarabine that are commonly used for the treatment of leukaemia, through cleavage of their triphosphorylated forms. Hence, SAMHD1 inhibitors are promising candidates for the sensitisation of leukaemia cells to nucleoside analogue-based therapy. Here, we investigated the effects of the cytosine analogue CNDAC, which has been proposed to be a SAMHD1 inhibitor, in the context of SAMHD1.
Methods: CNDAC was tested in 13 acute myeloid leukaemia (AML) cell lines, in 26 acute lymphoblastic leukaemia (ALL) cell lines, ten AML sublines adapted to various antileukaemic drugs, 24 single cell-derived clonal AML sublines, and primary leukaemic blasts from 24 AML patients. Moreover, 24 CNDAC-resistant sublines of the AML cell lines HL-60 and PL-21 were established. The SAMHD1 gene was disrupted using CRISPR/Cas9 and SAMHD1 depleted using RNAi, and the viral Vpx protein. Forced DCK expression was achieved by lentiviral transduction. SAMHD1 promoter methylation was determined by PCR after treatment of genomic DNA with the methylation-sensitive HpaII endonuclease. Nucleoside (analogue) triphosphate levels were determined by LC-MS/MS. CNDAC interaction with SAMHD1 was analysed by an enzymatic assay and by crystallisation.
Results: Although the cytosine analogue CNDAC was anticipated to inhibit SAMHD1, SAMHD1 mediated intrinsic CNDAC resistance in leukaemia cells. Accordingly, SAMHD1 depletion increased CNDAC triphosphate (CNDAC-TP) levels and CNDAC toxicity. Enzymatic assays and crystallisation studies confirmed CNDAC-TP to be a SAMHD1 substrate. In 24 CNDAC-adapted acute myeloid leukaemia (AML) sublines, resistance was driven by DCK (catalyses initial nucleoside phosphorylation) loss. CNDAC-adapted sublines displayed cross-resistance only to other DCK substrates (e.g. cytarabine, decitabine). Cell lines adapted to drugs not affected by DCK or SAMHD1 remained CNDAC sensitive. In cytarabine-adapted AML cells, increased SAMHD1 and reduced DCK levels contributed to cytarabine and CNDAC resistance.
Conclusion: Intrinsic and acquired resistance to CNDAC and related nucleoside analogues are driven by different mechanisms. The lack of cross-resistance between SAMHD1/ DCK substrates and non-substrates provides scope for next-line therapies after treatment failure.
(© 2021. The Author(s).)
Databáze: MEDLINE
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