Blood pressure-associated polymorphism controls ARHGAP42 expression via serum response factor DNA binding
Blood pressure-associated polymorphism controls ARHGAP42 expression via serum response factor DNA binding
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DOI：
10.1172/jci88899
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发表时间：
2017-02-01
影响因子：
15.9
通讯作者：
Mack, Christopher P.
中科院分区：
文献类型：
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作者：
Bai, Xue;Mangum, Kevin D.;Mack, Christopher P.
我们最近证明，在平滑肌细胞（SMC）中选择性表达Rho GTP酶激活蛋白ARHGAP 42通过抑制RhoA依赖性收缩力来控制血压，为ARHGAP 42基因内的血压相关位点提供了一种机制。本研究的目的是确定影响ARHGAP 42表达的多态性，并更好地评估ARHGAP 42在高血压发展中的作用。使用DNase I超敏反应方法和ENCODE数据，我们已经确定了一个包含ARHGAP 42 SNP rs604723的调控元件，该元件具有很强的SMC选择性，等位基因特异性活性。重要的是，CRISPR/Cas9介导的该元件在培养的人SMC中的缺失显著降低了内源性ARHGAP 42表达。DNA结合和转录测定表明，在rs604723的微小T等位基因变异增加了该片段的活性，通过促进血清反应转录因子结合到一个隐蔽的顺式元件。ARHGAP 42的表达通过细胞拉伸和1-磷酸鞘氨醇以RhoA依赖性方式增加，并且ARHGAP 42的缺失增强了用DOCA盐处理的小鼠中的高血压的进展。我们对一组未经治疗的临界性高血压患者的分析表明，ARHGAP 42基因型在高血压风险方面具有重要意义。总之，我们的数据增加了对控制血压的遗传机制的了解，并为个体化抗高血压治疗提供了潜在的靶点。
We recently demonstrated that selective expression of the Rho GTPase-activating protein ARHGAP42 in smooth muscle cells (SMCs) controls blood pressure by inhibiting RhoA-dependent contractility, providing a mechanism for the blood pressure-associated locus within the ARHGAP42 gene. The goals of the current study were to identify polymorphisms that affect ARHGAP42 expression and to better assess ARHGAP42's role in the development of hypertension. Using DNase I hypersensitivity methods and ENCODE data, we have identified a regulatory element encompassing the ARHGAP42 SNP rs604723 that exhibits strong SMC-selective, allele-specific activity. Importantly, CRISPR/Cas9-mediated deletion of this element in cultured human SMCs markedly reduced endogenous ARHGAP42 expression. DNA binding and transcription assays demonstrated that the minor T allele variation at rs604723 increased the activity of this fragment by promoting serum response transcription factor binding to a cryptic cis-element. ARHGAP42 expression was increased by cell stretch and sphingosine 1-phosphate in a RhoA-dependent manner, and deletion of ARHGAP42 enhanced the progression of hypertension in mice treated with DOCA-salt. Our analysis of a well-characterized cohort of untreated borderline hypertensive patients suggested that ARHGAP42 genotype has important implications in regard to hypertension risk. Taken together, our data add insight into the genetic mechanisms that control blood pressure and provide a potential target for individualized antihypertensive therapies.