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

  • 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.

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Author:Tamara RothenburgerORCiD, Dominique Jeanette ThomasORCiDGND, Yannick Schreiber, Paul Robin Wratil, Tamara Pflantz, Kirsten M. KnechtORCiD, Katie Digianantonio, Joshua Temple, Constanze SchneiderGND, Hanna-Mari Baldauf, Katie-May McLaughlin, Florian RothweilerGND, Berna Bilen, Samira Farmand, Denisa BojkovaORCiDGND, Rui CostaORCiDGND, Nerea Ferreirós BouzasORCiDGND, Gerd GeisslingerORCiDGND, Thomas OellerichORCiD, Yong XiongORCiD, Oliver Till KepplerORCiDGND, Mark N. WassORCiD, Martin MichaelisORCiDGND, Jindrich CinatlORCiDGND
Parent Title (English):Journal of experimental & clinical cancer research
Publisher:Springer ; BioMed Central
Place of publication:Berlin ; Heidelberg ; London
Document Type:Article
Date of Publication (online):2021/10/12
Date of first Publication:2021/10/12
Publishing Institution:Universitätsbibliothek Johann Christian Senckenberg
Release Date:2022/01/11
Tag:Acquired resistance; Acute lymphoblastic leukemia; Acute myeloid leukemia; CNDAC; DCK; Intrinsic resistance; Leukemia; SAMHD1; Sapacitabine
Issue:art. 317
Page Number:19
First Page:1
Last Page:19
The study was supported by the Frankfurter Stiftung für krebskranke Kinder and the Hilfe für krebskranke Kinder Frankfurt e.V. Open Access funding enabled and organized by Projekt DEAL.
The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
Dewey Decimal Classification:6 Technik, Medizin, angewandte Wissenschaften / 61 Medizin und Gesundheit / 610 Medizin und Gesundheit
Licence (German):License LogoCreative Commons - Namensnennung 4.0