Non-Coding Variants of Angiotensinogen Gene and Hypertension

血管紧张素原基因的非编码变异与高血压

• 批准号: 9197334
• 负责人: ASHOK KUMAR
• 金额: $ 61.5万
• 依托单位: NEW YORK MEDICAL COLLEGE
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-01 至 2019-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9197334
• 关键词: 3' Untranslated Regions; Address; Adipose tissue; Alleles; Angiotensin II; Angiotensinogen; Animals; Base Sequence; Binding; Binding Sites; Biological Assay; Blood Pressure; Carbohydrates; Chromatin; CpG dinucleotide; DNA; DNA Methylation; DNA analysis; Elements; Enhancers; Exons; Family; Fatty acid glycerol esters; Female; GTP-Binding Protein alpha Subunits, Gs; Genes; Genome; Haplotypes; Heart failure; Hepatocyte; Human; Hypertension; Individual; Inflammation; Introns; Kidney; Kidney Failure; Knock-in; Liver; Molecular; Mus; Myocardial Infarction; Nucleotides; Organ; Oxidative Stress; Play; Process; Proteins; Renin; Renin-Angiotensin System; Reporter; Risk Factors; Role; Sodium Chloride; Stroke; Tissues; Transcriptional Regulation; Transfection; Transgenic Animals; Transgenic Mice; Untranslated RNA; Variant; Vascular Diseases; base; blood pressure regulation; chromatin immunoprecipitation; chromosome conformation capture; genome wide association study; high salt diet; inflammatory marker; kidney cell; male; promoter; public health relevance; transcription factor; western diet

 DESCRIPTION (provided by applicant): Hypertension is a serious risk factor for myocardial infarction, heart failure, vascular disease, stroke, and renal failure. The renin-angiotensin system (RAS) plays an important role in the regulation of blood pressure (BP). Recent genome wise association studies (GWAS) have shown that an A/G SNP (rs2004776) located at +1164 in intron-I of the human angiotensinogen (hAGT) gene is associated with hypertension. The nucleotide sequence of hAGT gene containing +1164A allele has stronger homology with HNF-3 binding site as compared to +1164G. HNF3 family belongs to "pioneer" transcription factors whose binding to promoters and enhancers enables chromatin access for other tissue-specific transcription factors. SNPs in the promoter and intron I of the hAGT gene can be divided in 2 major haplotypes: haplotype-I (containing +1164A) and II (containing +1164G). Nucleotide variants in haplotype-I bind more strongly to transcription factors as compared to haplotype-II and reporter construct containing haplotype-I has increased promoter activity as compared to haplotype-II on transient transfection. Transgenic mice containing either haplotype-I or II of the hAGT gene were generated by knock-in approach at the HPRT locus. Preliminary studies have shown that: (a) CpG dinucleotides in the hAGT promoter of TG mice containing haplotype- I are hypo-methylated and accessible to transcription factors as compared to haplotype-II, (b) male TG mice containing haplotype-I have increased basal and GR induced expression of the hAGT gene in liver, kidney and fat as compared to haplotype-II, (c) chromatin from liver and kidney of transgenic animals containing haplotype-I binds more strongly to HNF3β, C/EBPβ, STAT-3 and GR as compared to haplotype-II, (d) double TG mice containing hREN gene and haplotype-I of the hAGT gene have increased BP as compared to haplotype-II which is further increased by Western diet containing high carbohydrate: high fat: high salt diet. In the present application, th role of SNP at +1164 and other cis-acting DNA elements in intron-I on transcriptional regulation of the hAGT gene will be analyzed by DNA methylation assay, chromosome conformation capture (3C) assay, ChIP assay and transient transfection assay. The effect of Western diet on the expression of the hAGT gene, blood pressure regulation, and end organ damage will be analyzed using male/female double transgenic mice containing hREN gene and either haplotype-I or haplotype-II of the hAGT gene.

 描述（由适用提供）：高血压是心肌梗塞，心力衰竭，血管疾病，中风和肾衰竭的严重危险因素。肾素 - 血管紧张素系统（RAS）在血压调节（BP）中起重要作用。最近的基因组明智的关联研究（GWAS）表明，人血管紧张素原（HAGT）基因内含子I中的A/G SNP（RS2004776）与高血压有关。与 +1164g相比，含有 +1164a等位基因的HAGT基因的核厄尔序序列与HNF-3结合位点具有很强的同源性。 HNF3家族属于“先驱”转录因子，其与启动子的结合和增强子可以使染色质访问其他组织特异性转录因子。 HAGT基因的启动子和内含子I中的SNP可以分为两种主要的单倍型：单倍型-I（包含 +1164a）和II（包含 +1164G）。与单倍型-II和含有单倍型-I的报告构建体相比，单倍型-I中的核苷酸变体与转录因子更强地结合，与瞬态翻译中的单倍型-II相比，启动子活性增加了启动子活性。通过HPRT基因座的敲入方法生成了含有HAGT基因的单倍型I或II的转基因小鼠。初步研究表明：（a）与单型II相比，与转录因子相比，含有单倍型I的TG小鼠的hagt启动子中的CpG二核苷酸是含有甲基化的，（b）含有单型型的雄性TG小鼠的基本和gr诱导的hagi hagi in gene in liver in liver in liver in liver（b）neplotype-i的雄性TG小鼠的表达增加了。与单倍型-II相比，来自含有单倍型-I的转基因动物的肝脏和肾脏的染色质与HNF3β，C/EBPβ，Stat-3和GR更强烈地结合，（D）含有HAGT基因的Hagt Gene的hagt型和Hagt Gene的HAGT型与Hagplot型相比增加了Hyplotype-ii II的富度效果，这是HYPLOT型的高效率，这是富含Hyploty型的高含量，这是HYPLOT型的高效率，这是Hyplot型的高效率，这是Hyplot型的高效果。盐饮食。在介绍应用中，将通过DNA甲基化测定法，染色体会议捕获（3C）测定，CHIP分析和瞬时转换测定法分析SNP在内含子I中的SNP和内含子I中其他顺式作用DNA元素的作用。西方饮食对HAGT基因表达，血压调节和最终器官损伤的影响将使用HAGT基因的男性/女性双重转基因小鼠以及单倍型I或单倍型-II进行分析。