Subtype-specific 3D genome alteration in acute myeloid leukaemia

• Published in: Nature (London) 2022-11, Vol.611 (7935), p.387-398
• Main Authors: Xu, Jie, Song, Fan, Lyu, Huijue, Kobayashi, Mikoto, Zhang, Baozhen, Zhao, Ziyu, Hou, Ye, Wang, Xiaotao, Luan, Yu, Jia, Bei, Stasiak, Lena, Wong, Josiah Hiu-yuen, Wang, Qixuan, Jin, Qi, Jin, Qiushi, Fu, Yihao, Yang, Hongbo, Hardison, Ross C., Dovat, Sinisa, Platanias, Leonidas C., Diao, Yarui, Yang, Yue, Yamada, Tomoko, Viny, Aaron D., Levine, Ross L., Claxton, David, Broach, James. R., Zheng, Hong, Yue, Feng
• Format: Article
• Language: English
• Subjects: 38; 45/15; 45/23; 631/67/1990/283/1897; 631/67/69; Acute myeloid leukemia; Bisulfite; Chromatin; Chromatin - genetics; CRISPR; CRISPR-Cas Systems; Deoxyribonucleic acid; DNA; DNA (Cytosine-5-)-Methyltransferases; DNA Methylation; DNMT1 protein; Enhancer Elements, Genetic; Enhancers; Epigenetics; Gene expression; Gene Expression Regulation, Leukemic; Gene sequencing; Gene Silencing; Genetic engineering; Genome, Human - genetics; Genomes; Humanities and Social Sciences; Humans; Interference; Leukemia; Leukemia, Myeloid, Acute - genetics; Malignancy; multidisciplinary; Mutation; Promoter Regions, Genetic; Regulatory sequences; Reproducibility of Results; Science; Science (multidisciplinary); Sequence Analysis; Silencers; Transcription factors; Whole genome sequencing
• ISSN: 0028-0836; 1476-4687
• DOI: 10.1038/s41586-022-05365-x

Acute myeloid leukaemia (AML) represents a set of heterogeneous myeloid malignancies, and hallmarks include mutations in epigenetic modifiers, transcription factors and kinases 1 – 5 . The extent to which mutations in AML drive alterations in chromatin 3D structure and contribute to myeloid transformation is unclear. Here we use Hi-C and whole-genome sequencing to analyse 25 samples from patients with AML and 7 samples from healthy donors. Recurrent and subtype-specific alterations in A/B compartments, topologically associating domains and chromatin loops were identified. RNA sequencing, ATAC with sequencing and CUT&Tag for CTCF, H3K27ac and H3K27me3 in the same AML samples also revealed extensive and recurrent AML-specific promoter–enhancer and promoter–silencer loops. We validated the role of repressive loops on their target genes by CRISPR deletion and interference. Structural variation-induced enhancer-hijacking and silencer-hijacking events were further identified in AML samples. Hijacked enhancers play a part in AML cell growth, as demonstrated by CRISPR screening, whereas hijacked silencers have a downregulating role, as evidenced by CRISPR-interference-mediated de-repression. Finally, whole-genome bisulfite sequencing of 20 AML and normal samples revealed the delicate relationship between DNA methylation, CTCF binding and 3D genome structure. Treatment of AML cells with a DNA hypomethylating agent and triple knockdown of DNMT1 , DNMT3A and DNMT3B enabled the manipulation of DNA methylation to revert 3D genome organization and gene expression. Overall, this study provides a resource for leukaemia studies and highlights the role of repressive loops and hijacked cis elements in human diseases. Extensive genomic analyses of the chromatin architecture in acute myeloid leukaemia reveals several characteristics, including subtype-specific distal enhancers and silencers, that may represent new anticancer therapeutic targets.