Physiologic profiling of sGC genetic variants

描述（由申请人提供）：高血压是心血管、脑血管和肾脏疾病的强风险因素。虽然具有高度遗传性，但一般人群中血压变化的遗传基础定义不清。在初步工作中，在GUCY 1A 3/GUCY 1B 3基因座鉴定出与血压相关的常见遗传变异，该基因座编码可溶性鸟苷酸环化酶的α 1和β 1亚基，该酶在被一氧化氮激活时合成cGMP，并在血管系统中大量表达。在多达200，000名欧洲血统的个体中，58%的人口具有两个BP升高等位基因的拷贝。拟议研究计划的总体目标是确定与血压相关的GUCY 1A 3/GUCY 1B 3遗传变异（或密切相关的变异）是否调节可溶性鸟苷酸环化酶α 1和/或β 1亚基的表达。在目的1中，我们试图确定BP与GUCY 1A 3/GUCY 1B 3位点的SNP的关联是否可能由sGC活性的差异决定。将对具有0或2个BP升高等位基因拷贝的个体施用吸入型一氧化氮，并将确定基因型对NO吸入增加血浆cGMP水平的能力的影响。在平行实验中，将在基因型之间比较血小板可溶性鸟苷酸环化酶亚单位mRNA和蛋白水平，以及酶活性和NO抑制聚集。在目标2中，将遗传变体引入含有人GUCY 1A 3/GUCY 1B 3基因座的细菌人工染色体中。将含有主要和次要等位基因的克隆引入血管平滑肌细胞，并比较可溶性鸟苷酸环化酶α 1和β 1亚基的表达。研究人员提出的使用详细的生理表型和DNA序列变异的分子表征有望产生对GUCY 1A 3和GUCY 1B 3基因调控的基本见解，并开始更好地了解一氧化氮可溶性鸟苷酸环化酶-cGMP信号传导在普通人群血压调节中的作用。

DESCRIPTION (provided by applicant): High blood pressure is a strong risk factor for cardiovascular, cerebrovascular, and kidney disease. Although highly heritable, the genetic basis of blood pressure variation in the general population has been poorly-defined. In preliminary work, a common genetic variant associated with blood pressure was identified at the GUCY1A3/GUCY1B3 locus, which encodes the alpha1 and beta1 subunit of soluble guanylate cyclase, an enzyme which synthesizes cGMP when activated by nitric oxide and which is abundantly expressed in the vasculature. Among up to 200,000 individuals of European ancestry, 58% of the population has two copies of a BP-raising allele. The overall goal of the proposed research program is to ascertain whether the GUCY1A3/GUCY1B3 genetic variant associated with blood pressure (or a closely linked variant) modulates expression of soluble guanylate cyclase alpha1 and/or beta1 subunits. In Aim 1, we seek to establish whether the association of BP with the SNP at the GUCY1A3/GUCY1B3 locus is likely to be determined by a difference in sGC activity. Inhaled nitric oxide will be administered to individuals with 0 or 2 copies of the BP-raising allele, and the impact of genotype on the ability of NO inhalation to increase plasma cGMP levels will be determined. In parallel experiments, platelet soluble guanylate cyclase subunit mRNA and protein levels, as well as enzyme activity and NO- inhabitable aggregation will be compared between genotypes. In Aim 2, genetic variants will be introduced into bacterial artificial chromosomes containing the human GUCY1A3/GUCY1B3 locus. Clones containing major and minor alleles will be introduced in vascular smooth muscle cells, and expression of the soluble guanylate cyclase alpha1 and beta1 subunits will be compared. The investigators' proposed use of detailed physiologic phenotyping and molecular characterization of the DNA sequence variation promises to yield fundamental insights into regulation of the GUCY1A3 and GUCY1B3 genes and to begin to better understand the role of nitric oxide-soluble guanylate cyclase-cGMP signaling in blood pressure regulation in the general population.