Fig. 1: Dnmt3aR882H/+ HSPC shows self-renewal phenotype, enhanced BM repopulation and progression to MPN/AML.
From: Mitochondrial metabolism sustains DNMT3A-R882-mutant clonal haematopoiesis
a, Structure of the Dnmt3aR882H conditional allele. b, RNA sequencing reads from Dnmt3a+/+ and Dnmt3aR882H/+ HSPCs were aligned to mouse exons and human exon 23 with DNMT3A-R882H mutation. Sanger sequencing was performed on cDNA of Dnmt3aR882H/+ amplified with primers detecting human exon 23. Similar data were observed for n = 3 mice per genotype. c, Serial replating of BM-derived colonies from Dnmt3a+/+ (n = 7 mice) and Dnmt3aR882H/+ (n = 8 mice). d, Schematic representation of BM competitive transplant strategy of CD45.2-WT and CD45.2-Dnmt3aR882H/+ transplanted together with CD45.1 competitor. e, Proportion of Dnmt3a+/+ (n = 5 mice) and Dnmt3aR882H/+ (n = 4 mice) cells in PB post-transplant. f,g, Fluorescence-activated cell sorting plots of LT-HSC (Lin−ve, Sca1+, c-Kit+, Cd48−Cd150+) and proportion of Dnmt3a+/+ (n = 5 mice) and Dnmt3aR882H/+ (n = 4 mice) transplanted cells in LT-HSC (f) with quantification (g). h, Kaplan–Meier survival curves for Dnmt3aR882H/+ (n = 35) and control (n = 31) mice; P by log-rank (Mantel–Cox) test. i, Histopathological diagnoses of moribund mice. Bars depict cancer/normal diagnosis per genotype/total mice with available histology data (n = 44 mice). j, Characteristic histology from one mouse with MPN/AML. Myeloid cell/AML infiltration in the liver (Li) and blasts in the setting of myeloid hyperplasia and a hypercellular marrow are shown. Similar data were observed for 13 Dnmt3aR882H/+ mice. w, week. Scale bars, 300 µm for inset (Li), 50 µm for others. In c–e, g and i, the mean ± s.d. is shown; P by two-sided t-test for comparisons between Dnmt3aR882H/+ and WT. Schematics in a,d were created using BioRender (https://biorender.com).
