Disruption of CTCF Boundary at HOXA Locus Promote BET Inhibitors’ Therapeutic Sensitivity in Acute Myeloid Leukemia

• Published in: Stem cell reviews and reports 2020-12, Vol.16 (6), p.1280-1291
• Main Authors: Zha, Jie, Lai, Qian, Deng, Manman, Shi, Pengcheng, Zhao, Haijun, Chen, Qinwei, Wu, Hua, Xu, Bing
• Format: Article
• Language: English
• Subjects: Acute myeloid leukemia; Biomedical and Life Sciences; Biomedical Engineering and Bioengineering; Cell Biology; Chromatin; CRISPR; DNA-directed RNA polymerase; Drug resistance; Gene expression; Genome editing; Genomes; HOX gene; HOXA gene; HOXA9 gene; Life Sciences; MAP kinase; Myeloid leukemia; Regenerative Medicine/Tissue Engineering; Ribonucleic acid; RNA; Signal transduction; Stem Cells
• ISSN: 2629-3269; 2629-3277
• DOI: 10.1007/s12015-020-10057-y

Both HOX gene expression and CTCF regulation have been well demonstrated to play a critical role in regulating maintenance of leukemic stem cells (LSCs) that are known to be resistant to BET inhibitor (BETi). To investigate the regulatory role of CTCF boundary in aberrant HOX gene expression and the therapeutic sensitivity of BETi in AML, we employed CRISPR-Cas9 genome editing technology to delete 47 base pairs of the CTCF binding motif which is located between HOXA7 and HOXA9 genes (CBS7/9) in different subtypes of AML with either MLL -rearrangement or NPM1 mutation. Our results revealed that HOXA9 is significantly downregulated in response to the CBS7/9 deletion. Moreover, CBS7/9 boundary deletion sensitized the BETi treatment reaction in both MOLM-13 and OCI-AML3 cells. To further examine whether BETi therapeutic sensitivity in AML is depended on the expression level of the HOXA9 gene, we overexpressed the HOXA9 in the CBS7/9 deleted AML cell lines, which can rescue and restore the resistance to BETi treatment of the CBS7/9 KO cells by activating MAPK signaling pathway. Deletion of CBS7/9 specifically decreased the recruitment of BRD4 and RNA pol II to the posterior HOXA genes, in which, a transcription elongation factor ELL3 is the key factor in regulating HOXA gene transcription monitored by CBS7/9 chromatin boundary. Thus, disruption of CBS7/9 boundary perturbs HOXA9 transcription and regulates BETi sensitivity in AML treatment. Moreover, alteration of CTCF boundaries in the oncogene loci may provide a novel strategy to overcome the drug resistance of LSCs. Graphical abstract