TY  - JOUR
A1  - Rothenburger, Tamara
A1  - Thomas, Dominique Jeanette
A1  - Schreiber, Yannick
A1  - Wratil, Paul Robin
A1  - Pflantz, Tamara
A1  - Knecht, Kirsten M.
A1  - Digianantonio, Katie
A1  - Temple, Joshua
A1  - Schneider, Constanze
A1  - Baldauf, Hanna-Mari
A1  - McLaughlin, Katie-May
A1  - Rothweiler, Florian
A1  - Bilen, Berna
A1  - Farmand, Samira
A1  - Bojkova, Denisa
A1  - Costa, Rui
A1  - Ferreirós Bouzas, Nerea
A1  - Geisslinger, Gerd
A1  - Oellerich, Thomas
A1  - Xiong, Yong
A1  - Keppler, Oliver Till
A1  - Wass, Mark N.
A1  - Michaelis, Martin
A1  - Cinatl, Jindrich
T1  - Differences between intrinsic and acquired nucleoside analogue resistance in acute myeloid leukaemia cells
T2  - Journal of experimental & clinical cancer research
N2  - 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.
KW  - Leukemia
KW  - Acute myeloid leukemia
KW  - Acute lymphoblastic leukemia
KW  - CNDAC
KW  - Sapacitabine
KW  - SAMHD1
KW  - DCK
KW  - Intrinsic resistance
KW  - Acquired resistance
Y1  - 2021
UR  - http://publikationen.ub.uni-frankfurt.de/frontdoor/index/index/docId/64010
UR  - https://nbn-resolving.org/urn:nbn:de:hebis:30:3-640108
SN  - 1756-9966
N1  - 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.
N1  - 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.
VL  - 40
IS  - art. 317
SP  - 1
EP  - 19
PB  - Springer ; BioMed Central
CY  - Berlin ; Heidelberg ; London
ER  -