Abstract
Constitutively activated B cell receptor (BCR) signaling is a primary biological feature of chronic lymphocytic leukemia (CLL). The biological events controlled by BCR signaling in CLL are not fully understood and need investigation. To make inroads we obtained blood samples from CLL patients before and after Bruton’s tyrosine kinase inhibitors (BTKi) treatment and used them to study BCR signaling regulated genes. Here, by analysis of the chromatin states and gene expression profiles of CLL B cells from patients before and after BTKi ibrutinib treatment, we show that BTKi treatment leads to a decreased expression of APOBEC3 family genes in an enhancer regulation dependent manner. BTKi treatment reduces enrichment of enhancer markers (H3K4me1, H3K27ac) and chromatin accessibility at putative APOBEC3 enhancers. CRISPR-Cas9 directed deletion or inhibition of the putative APOBEC3 enhancers leads to reduced APOBEC3 expression. We further find that transcription factor NFATc1 couples BCR signaling with the APOBEC3 enhancer activity to control APOBEC3 expression. Importantly, enhancer regulated APOBEC3 expression contributes to replication stress in malignant B cells. We also demonstrate a novel mechanism for BTKi suppression of APOBEC3 expression via direct enhancer regulation in a NFATc1 dependent manner, implicating BCR signaling as a potential regulator of leukemic genomic instability.
Key points
BCR signaling pathway regulates APOBEC3 expression via direct enhancer regulation.
AOPEBC3 enhancers are involved in the process of DNA replication stress, implicating a potential role in B cell genomic instability and CLL evolution
Competing Interest Statement
NEK Advisory Board for: Abbvie, Astra Zeneca, Behring, Cytomx Therapy, Dava oncology, Janssen, Juno Theraputics, Oncotracker, Pharmacyclics and Targeted Oncology. DSMC (Data Safety Monitoring Committee) for: Agios Pharm, AstraZeneca, BMS/Celgene, Cytomx Therapeutics, Janssen, Morpho-sys, Rigel. Research funding from: Abbvie, Acerta Pharma, Bristol Meyer Squib, Celgene, Genentech, MEI Pharma, Pharmacyclics, Sunesis, TG Therapeutics, Tolero Pharmaceuticals. SAP: Research funding has been provided to the institution from Pharmacyclics, Janssen, AstraZeneca, TG Therapeutics, Merck, AbbVie, and Ascentage Pharma for clinical studies in which Sameer A. Parikh is a principal investigator. SAP has also participated in Advisory Board meetings of Pharmacyclics, AstraZeneca, Genentech, GlaxoSmithKline, Adaptive Health, Adaptive Biotechnologies, and AbbVie (he was not personally compensated for his participation).