Physiologic Profiling of sGC Genetic Variants

sGC 遗传变异的生理分析

• 批准号: 8866446
• 负责人: Christopher Holmes Newton-Cheh
• 金额: $ 56.17万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-08-05 至 2016-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8866446
• 关键词: 3' Flanking Region; Alleles; Bacterial Artificial Chromosomes; Biological Assay; Blood Platelets; Blood Pressure; Blood Vessels; Breathing; Candidate Disease Gene; Cardiovascular Diseases; Cerebrovascular Disorders; Chronic Kidney Failure; Clinical; Code; Complex; Cyclic GMP; DNA Resequencing; DNA Sequence; Development; Diastolic blood pressure; Enzymes; European; Funding; GUCY1A3 gene; GUCY1B3 gene; Gene Deletion; Gene Expression; General Population; Genes; Genetic; Genetic Determinism; Genetic Variation; Genotype; Goals; Health; Heart failure; Heritability; Homozygote; Human; Hypertension; Individual; Intervention; Introns; Investigation; Kidney; Kidney Diseases; Link; Marketing; Messenger RNA; Methods; Minor; Mus; Mutation; Myocardial Infarction; Natriuretic Peptides; Nitric Oxide; Nucleotides; PDE3A gene product; Peptide Receptor; Physiological; Physiology; Plasma; Platelet Activation; Platelet aggregation; Population; Prevalence; Proteins; Pulmonary artery structure; Rattus; Recruitment Activity; Regulation; Research; Research Personnel; Risk Factors; Role; Signal Pathway; Signal Transduction; Single Nucleotide Polymorphism; Smooth Muscle Myocytes; Sodium Chloride; Soluble Guanylate Cyclase; Source; Stroke; System; Transcript; Transfection; Translating; Translations; Variant; Vasodilation; Work; base; blood pressure regulation; enzyme activity; experience; genetic analysis; genetic variant; genome wide association study; guanylate cyclase 1; healthy volunteer; human NOS3 protein; improved; inhaled nitric oxide; insight; molecular phenotype; novel; programs; research study; response; trait

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.

描述（申请人提供）：高血压是心血管、脑血管和肾脏疾病的强烈危险因素。尽管具有高度遗传性，但普通人群血压变异的遗传基础尚不清楚。在初步工作中，在 GUCY1A3/GUCY1B3 基因座上发现了一种与血压相关的常见遗传变异，该变异编码可溶性鸟苷酸环化酶的 α1 和 β1 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 200,000 名欧洲血统的个体中，58% 的人拥有两个升高血压的等位基因。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 α1 和/或 β1 亚基的表达。在目标 1 中，我们试图确定 BP 与 GUCY1A3/GUCY1B3 基因座上的 SNP 的关联是否可能由 sGC 活性的差异决定。将向具有 0 或 2 个升高血压等位基因拷贝的个体施用吸入一氧化氮，并确定基因型对吸入一氧化氮提高血浆 cGMP 水平的能力的影响。在平行实验中，将比较基因型之间的血小板可溶性鸟苷酸环化酶亚基 mRNA 和蛋白质水平，以及酶活性和 NO-可居住聚集。在目标 2 中，遗传变异将被引入包含人类 GUCY1A3/GUCY1B3 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α1和β1亚基的表达。研究人员提议使用详细的生理表型分析和 DNA 序列变异的分子表征，有望对 GUCY1A3 和 GUCY1B3 基因的调节产生基本的了解，并开始更好地了解一氧化氮可溶性鸟苷酸环化酶 -cGMP 信号在普通人群血压调节中的作用。