Transcriptional regulation of human angiotensin receptor

人血管紧张素受体的转录调控

基本信息

批准号：
7658752
负责人：
ASHOK KUMAR
金额：
$ 38.35万
依托单位：
NEW YORK MEDICAL COLLEGE
依托单位国家：
美国
项目类别：
财政年份：
2006
资助国家：
美国
起止时间：
2006-07-01 至 2011-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7658752
关键词：
5&apos Flanking Region Adrenal Glands Aldosterone Angiotensin II Angiotensin Receptor Angiotensins Bacterial Artificial Chromosomes Binding Blood Pressure Blood Vessels CCAAT-Enhancer-Binding Proteins Cardiovascular Diseases Caucasians Caucasoid Race Cells DNA Down-Regulation E-Box Elements Electrophoretic Mobility Shift Assay Estrogens Etiology Female Gene Expression Genes Genetic Polymorphism Genetic Transcription Genomics Glucocorticoids Haplotypes Heart failure Homeostasis Human Hyperglycemia Hypertension Hypoxia In Vitro Incidence Inflammation Interleukin-6 Kidney Failure Lead Liquid substance Messenger RNA Molecular Mus Myocardial Infarction Nucleosides Oligonucleotides Pathogenesis Physiological Play Postmenopause Production Receptor Gene Receptor, Angiotensin, Type 1 Renal function Renin-Angiotensin System Reporter Right-On Risk Factors Role Secondary to Single Nucleotide Polymorphism Smooth Muscle Myocytes Stroke System Testing Transcriptional Regulation Transfection Transgenic Mice Translations Variant Vascular Diseases Vascular Smooth Muscle Woman base factor C in vivo Model male normotensive promoter response transcription factor transcription factor USF vasoconstriction vpr Genes

项目摘要

DESCRIPTION (provided by applicant): The octapeptide, angiotensin-ll (Ang II) is one of the most potent vasoactive substances known and regulates a variety of physiological responses, including fluid homeostasis, aldosterone production, renal function and contraction of vascular smooth muscle (VSM). Over-expression of the angiotensin receptor-1 (AT1R) gene produces hypertension especially in female transgenic mice. These studies have suggested that increased transcription of the hAT1 R gene may lead to hypertension. We have therefore analyzed the role of single nucleotide polymorphisms (SNPs) in the 5' flanking region of the hAT1R gene in hypertension. We have found that hAT1 R gene promoter has a haplotype block of at least five SNPs consisting of T/A at - 777, T/G at -680, A/C at -214, G/C at -213, and A/G at -119. Our studies have shown that variants -777T, - 680T, -214A, -213G, and -119A always occur together (creating haplotype-l containing TTAGA and haplotype-ll containing AGCCG respectively). Our studies have shown that haplotype-l of the hAT1R gene is associated with hypertension in Caucasian women and transient transfection of reporter construct containing haplotype-l of the hAT1R gene has increased promoter activity in adrenal cortical cells and VSMC as compared to haplotype-ll. Our gel shift assays have shown that: (a) transcription factor C/EBP binds more strongly to an oligonucleotide containing -119A (haplotype-l) as compared to -119G (haplotype- ll) and (b) transcription factor USF (which binds to an E-box motif CANNTG) binds strongly to an oligonucleotide containing nucleoside A and G at -214 and -213 (haplotype-l) as compared to the same oligonucleotide containing nucleoside C at -213 and -214 (haplotype-ll). We will therefore analyze the effect of haplotypes -I and II on transcriptional regulation of the hAT1R gene in an in-vitro system using adrenocortical and vascular smooth muscle cells, examine the effect of haplotypes-l and II on hAT1 R mRNA level and on blood pressure using male and female transgenic mice, and examine the effect of IL-6, glucocorticoids, and estrogens on hAT1R mRNA level in transgenic mice containing haplotype I and II of the hAT1 R gene, and to correlate potential changes in mRNA levels with blood pressure in transgenic mice.

描述（由申请人提供）：八肽，血管紧张素-LL（ANG II）是已知的最有效的血管活性物质之一，可以调节各种生理反应，包括液体稳态，醛固酮产生，肾功能，血管平滑肌的血管平滑肌（VSM）。血管紧张素受体1（AT1R）基因的过表达会产生高血压，尤其是在雌性转基因小鼠中。这些研究表明，HAT1 r基因的转录增加可能导致高血压。因此，我们已经分析了HAT1R基因的5'侧翼区域中单核苷酸多态性（SNP）在高血压中的作用。我们发现HAT1 r基因启动子具有至少五个SNP的单倍型块，由T/A AT -777，T/G at -680，-680，A/C at -214，-214，g/c at -213和-119。我们的研究表明，变体-777t，-680t，-214a，-213g和-119a总是一起出现的（分别创建包含ttaga和单倍型-LL的单倍型-l，分别包含AGCCG）。我们的研究表明，与单倍型相比，HAT1R基因的单倍型L与高加索妇女的高血压和含有HAT1R基因的单倍型L的报告构建体的瞬时转染具有增加的启动子活性。我们的凝胶转移测定法表明：（a）转录因子C/EBP与含-119a（habletype -l）的寡核苷酸更强地结合，与-119g（单型-LL）相比-213（单倍型-L）与含有-213和-214（单倍型-ll）的相同寡核苷酸C相比。 We will therefore analyze the effect of haplotypes -I and II on transcriptional regulation of the hAT1R gene in an in-vitro system using adrenocortical and vascular smooth muscle cells, examine the effect of haplotypes-l and II on hAT1 R mRNA level and on blood pressure using male and female transgenic mice, and examine the effect of IL-6, glucocorticoids, and estrogens on hAT1R mRNA level in含有HAT1 R基因的单倍型和II的转基因小鼠，并将mRNA水平的潜在变化与转基因小鼠的血压相关。