The Histone Demethylase JMJD3 Regulates Smooth Muscle Differentiation And Is Associated With Blood Pressure

Hypertension is associated with significant morbidity and mortality in the United States, and despite its prevalence an unmet need exists for more personalized targeted therapies. Blood pressure is regulated by environmental and genetic factors, and genome wide association studies (GWAS) have identified numerous genes in the human population associated with hypertension. Recently, one GWAS identified the single nucleotide polymorphism rs62059712 that is associated with systolic blood pressure in humans. rs62059712 is located upstream of the gene JMJD3, a histone demethylase that our lab has studied in a variety of disease contexts, and the rs62059712 major C allele (MAF 0.92) is associated with increased blood pressure in humans. Based on our analysis of data from the UCSC genome browser, we found that rs62059712 is located in a DNase Hypersensitive Site marked by H3K27Ac, suggesting it may regulate JMJD3 transcription. In luciferase assays, the fragment containing rs62059712 displayed transcription activity in smooth muscle cells (SMC), and the major C allele exhibited increased luciferase activity compared to the minor T allele in mouse aortic SMC (p<0.05). Notably, rs62059712 is in linkage disequilibrium with rs74480102, which did not display allele-specific transcription activity in SMC. siRNA knockdown of JMJD3 in mouse aortic SMC significantly decreased SMC-specific gene expression (p<0.05), and SMC marker gene expression was also reduced by treatment of SMC with the JMJD3 selective inhibitor GSK4J (p<0.05). Likewise, SMC contractile gene expression was reduced in aortas from SM22Cre;JMJD3 flox/flox mice harboring SMC-specific deletion of JMJD3. Further, angiotensin II increased JMJD3 expression in human aortic SMC. Finally, in chromatin immunoprecipitation assays in mouse aortic SMC, JMJD3 was significantly enriched at smooth muscle gene promoters (p<0.05). Our results identify the blood pressure-associated gene JMJD3 as a key epigenetic regulator of smooth muscle differentiation. Additionally, we show that the rs62059712 major C allele, which is associated with increased blood pressure, increases JMJD3 transcription activity, and likely expression, to drive increased downstream contractile smooth muscle gene expression.