The aberrant epigenome of DNMT3B-mutated ICF1 patient iPSCs is amenable to correction, with the exception of a subset of regions with H3K4me3-and/or CTCF-based epigenetic memory

被引：5

作者：

Krishnan, Varsha Poondi ^{[1
]}

Morone, Barbara ^{[1
]}

Toubiana, Shir ^{[2
,3
]}

Krzak, Monika ^{[4
,7
]}

Fioriniello, Salvatore ^{[1
]}

Della Ragione, Floriana ^{[1
,5
]}

Strazzullo, Maria ^{[1
]}

Angelini, Claudia ^{[4
]}

Selig, Sara ^{[2
,3
,6
]}

Matarazzo, Maria R. ^{[1
]}

机构：

[1] CNR, Inst Genet & Biophys Adriano Buzzati Traverso IGB, I-80131 Naples, Italy

[2] Technion, Rappaport Fac Med, Dept Genet & Dev Biol, IL-31096 Haifa, Israel

[3] Technion, Res Inst, IL-31096 Haifa, Israel

[4] CNR, Inst Appl Comp IAC Mauro Picone, I-80131 Naples, Italy

[5] IRCCS Ist Neurol Mediterraneo Neuromed, I-86077 Pozzilli, Isernia, Italy

[6] Rambam Hlth Care Campus, Lab Mol Med, IL-31096 Haifa, Israel

[7] Wellcome Sanger Inst, Cambridge CB10 1SA, England

来源：

GENOME RESEARCH | 2023年 / 33卷 / 02期

基金：

以色列科学基金会;

关键词：

DNA METHYLTRANSFERASE GENE; PLURIPOTENT STEM-CELLS; IMMUNODEFICIENCY SYNDROME; FACIAL ANOMALIES; METHYLATION; DNMT3B; TRANSCRIPTION; EXPRESSION; MUTATIONS; DIFFERENTIATION;

D O I：

10.1101/gr.276986.122

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Bi-allelic hypomorphic mutations in DNMT3B disrupt DNA methyltransferase activity and lead to immunodeficiency, centromeric instability, facial anomalies syndrome, type 1 (ICF1). Although several ICF1 phenotypes have been linked to abnormally hypomethylated repetitive regions, the unique genomic regions responsible for the remaining disease phenotypes remain largely uncharacterized. Here we explored two ICF1 patient-derived induced pluripotent stem cells (iPSCs) and their CRISPR-Cas9-corrected clones to determine whether DNMT3B correction can globally overcome DNA methylation defects and related changes in the epigenome. Hypomethylated regions throughout the genome are highly comparable between ICF1 iPSCs carrying different DNMT3B variants, and significantly overlap with those in ICF1 patient peripheral blood and lymphoblastoid cell lines. These regions include large CpG island domains, as well as promoters and enhancers of several lineage-specific genes, in particular immune-related, suggesting that they are premarked during early development. CRISPR-corrected ICF1 iPSCs reveal that the majority of phenotype-related hypomethylated regions reacquire normal DNA methylation levels following editing. However, at the most severely hypomethylated regions in ICF1 iPSCs, which also display the highest increases in H3K4me3 levels and/or abnormal CTCF binding, the epigenetic memory persists, and hypomethylation remains uncorrected. Overall, we demonstrate that restoring the catalytic activity of DNMT3B can reverse the majority of the aberrant ICF1 epigenome. However, a small fraction of the genome is resilient to this rescue, highlighting the challenge of reverting disease states that are due to genome-wide epigenetic perturbations. Uncovering the basis for the persistent epigenetic memory will promote the development of strategies to overcome this obstacle.

引用

页码：169 / 183

页数：15