Targeting of DNMT3A hotspot mutations in Clonal Hematopoiesis and Acute Myeloid Leukemia. Targeting of DNMT3A hotspot mutations in Clonal Hematopoiesis and Acute Myeloid Leukemia

Somatic mutations in DNA Methyltransferase 3A (DNMT3A) with a hotspot in exon 23 at Arginine 882 (DNMT3AR882mut) are the most frequent single mutations in clonal hematopoiesis. Here we analyze the expression of genes and endogenous retrovirus (ERV) sequences in a murine model carrying human DNMT3A-R882H mutation in one allele of the endogenous DNMT3A with respect to normal condition and azacitidine treatment. Overall design: Lineage- Sca1+ Kit + (LSK) sorted bone marrow cells from WT DNMT3A and mice habouring a human DNMT3A R882H mutant allele in the endogenous locus were subject to transcriptomic sequencing under normal and azazytidine treatment condition. Expression levels of genes as well as repetitive elements were investigated.

DNA甲基转移酶3A（DNA Methyltransferase 3A, DNMT3A）第23外显子精氨酸882位点存在热点突变的体细胞突变（DNMT3AR882mut）是克隆性造血（clonal hematopoiesis）中最常见的单基因突变。本研究针对携带内源性DNMT3A等位基因之一导入人源DNMT3A-R882H突变的小鼠模型，分别在正常培养条件与阿扎胞苷（azacitidine）处理下，分析其基因与内源性逆转录病毒（endogenous retrovirus, ERV）序列的表达情况。整体实验设计：从野生型（wild type, WT）DNMT3A小鼠以及内源性位点携带人源DNMT3A R882H突变等位基因的小鼠中分离谱系阴性、Sca1阳性、Kit阳性（LSK）骨髓分选细胞，在正常培养与阿扎胞苷处理条件下开展转录组测序，并对基因及重复序列的表达水平进行检测。