Physiologic Profiling of sGC Genetic Variants

描述（由申请人提供）：高血压是心脑血管和肾脏疾病的重要危险因素。虽然血压具有高度遗传性，但一般人群中血压变异的遗传基础尚未明确。在初步研究中，在GUCY1A3/GUCY1B3位点发现了一种与血压相关的常见遗传变异，该基因编码可溶性鸟苷酸环化酶的α 1和β 1亚基，这种酶在被一氧化氮激活时合成cGMP，并在脉管系统中大量表达。在多达20万欧洲人的祖先中，58%的人有两个升高bp的等位基因拷贝。拟议研究计划的总体目标是确定与血压相关的GUCY1A3/GUCY1B3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Aim 1中，我们试图确定BP与GUCY1A3/GUCY1B3位点SNP的关联是否可能由sGC活性的差异决定。对于携带0或2个bp升高等位基因拷贝的个体，将给予吸入一氧化氮，并确定基因型对吸入一氧化氮增加血浆cGMP水平的影响。在平行实验中，将比较不同基因型血小板可溶性鸟苷酸环化酶亚基mRNA和蛋白水平，以及酶活性和NO-可居住聚集。在目标2中，将遗传变异引入含有人类GUCY1A3/GUCY1B3位点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，比较可溶性鸟苷酸环化酶α 1和β 1亚基的表达。研究人员建议使用详细的生理表型和DNA序列变异的分子特征，有望对GUCY1A3和GUCY1B3基因的调控产生根本性的见解，并开始更好地理解一氧化氮可溶性鸟苷酸环化酶- cgmp信号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.