Gozdecka Malgorzata, Dudek Monika, Wen Sean, Gu Muxin, Stopforth Richard J, Rak Justyna, Damaskou Aristi, Grice Guinevere L, McLoughlin Matthew A, Bond Laura, Wilson Rachael, Giotopoulos George, Shanmugiah Vijaya Mahalingam, Bakar Rula Bany, Yankova Eliza, Cooper Jonathan L, Narayan Nisha, Horton Sarah J, Asby Ryan, Pask Dean C, Mupo Annalisa, Duddy Graham, Marando Ludovica, Georgomanolis Theodoros, Carter Paul, Ramesh Amirtha Priya, Dunn William G, Barcena Clea, Gallipoli Paolo, Yusa Kosuke, Petrovski Slavé, Wright Penny, Quiros Pedro M, Frezza Christian, Nathan James A, Kaser Arthur, Kar Siddhartha, Tzelepis Konstantinos, Mitchell Jonathan, Fabre Margarete A, Huntly Brian J P, Vassiliou George S
Cambridge Stem Cell Institute, University of Cambridge, Cambridge, UK.
Department of Haematology, University of Cambridge, Cambridge, UK.
Nature. 2025 Apr 16. doi: 10.1038/s41586-025-08980-6.
Somatic DNMT3A-R882 codon mutations drive the most common form of clonal haematopoiesis (CH) and are associated with increased acute myeloid leukaemia (AML) risk. Preventing expansion of DNMT3A-R882-mutant haematopoietic stem/progenitor cells (HSPCs) may therefore avert progression to AML. To identify DNMT3A-R882-mutant-specific vulnerabilities, we conducted a genome-wide CRISPR screen on primary mouse Dnmt3a HSPCs. Among the 640 vulnerability genes identified, many were involved in mitochondrial metabolism, and metabolic flux analysis confirmed enhanced oxidative phosphorylation use in Dnmt3a versus Dnmt3a (WT) HSPCs. We selected citrate/malate transporter Slc25a1 and complex I component Ndufb11, for which pharmacological inhibitors are available, for downstream studies. In vivo administration of SLC25A1 inhibitor CTPI2 and complex I inhibitors IACS-010759 and metformin suppressed post-transplantation clonal expansion of Dnmt3a, but not WT, long-term haematopoietic stem cells. The effect of metformin was recapitulated using a primary human DNMT3A-R882 CH sample. Notably, analysis of 412,234 UK Biobank participants showed that individuals taking metformin had a markedly lower prevalence of DNMT3A-R882-mutant CH, after controlling for potential confounders including glycated haemoglobin, diabetes and body mass index. Collectively, our data propose modulation of mitochondrial metabolism as a therapeutic strategy for prevention of DNMT3A-R882-mutant AML.
体细胞DNMT3A-R882密码子突变驱动了最常见的克隆性造血（CH）形式，并与急性髓系白血病（AML）风险增加相关。因此，阻止DNMT3A-R882突变的造血干/祖细胞（HSPCs）扩增可能避免进展为AML。为了确定DNMT3A-R882突变特异性的脆弱性，我们对原代小鼠Dnmt3a HSPCs进行了全基因组CRISPR筛选。在鉴定出的640个脆弱性基因中，许多基因参与线粒体代谢，代谢通量分析证实Dnmt3a突变型HSPCs相比于Dnmt3a（野生型）HSPCs对氧化磷酸化的利用增强。我们选择了有药理学抑制剂可用的柠檬酸/苹果酸转运蛋白Slc25a1和复合体I组分Ndufb11进行下游研究。在体内给予SLC25A1抑制剂CTPI2以及复合体I抑制剂IACS-010759和二甲双胍可抑制Dnmt3a突变型而非野生型长期造血干细胞移植后的克隆性扩增。使用原发性人类DNMT3A-R882 CH样本再现了二甲双胍的作用。值得注意地是，对412,234名英国生物银行参与者的分析表明，在控制了包括糖化血红蛋白、糖尿病和体重指数等潜在混杂因素后，服用二甲双胍的个体中DNMT3A-R882突变型CH的患病率显著更低。总体而言，我们的数据提出调节线粒体代谢作为预防DNMT3A-R882突变型AML的一种治疗策略。
