Regulatory RNA in Hypertension

项目总结 超过50%的高血压患者表现出对盐摄取的血压敏感性增加。 然而，事实证明，在全人口范围内减少食盐摄入量是困难的，这使得减少食盐摄入量变得更加重要 进一步了解盐敏感型高血压的发病机制，为开发新的盐敏感型高血压提供依据 干预措施。肾脏在包括盐分在内的高血压的发展中起着关键的生理作用。 敏感性高血压。 基因组中数以千计的基因编码长的非编码RNA(LncRNAs)。InncRNA可以相互作用 与其他核糖核酸或蛋白质结合并影响其功能。很少有人研究lncRNAs在 高血压。与大多数lncRNA不同，MALAT1(转移相关肺腺癌转录本1；MALAT1 在啮齿动物中)在许多物种中都是保守的，并在几种组织中以高丰度表达 包括肾脏，这表明MALAT1可能在生理上很重要。然而，MALAT1的S 生理学和病理生理学作用在很大程度上仍不清楚。 我们最近发现，肾脏miR-214-3p靶向并抑制内皮型一氧化氮合酶 (ENOS)，这在盐敏感型高血压大鼠模型的发展中起着重要作用。 可能还有人类。这得到了对人类序列变体和所有miRNA的系统分析的支持 前体，人肾活检标本中的小RNA深度测序，肾脏特异性抑制miR- 214-3P在Dahl SS大鼠和一个新产生的突变大鼠品系。Dahl SS大鼠是应用最广泛的模型 目的：研究人类盐敏感型高血压的分子机制。 我们已经获得了一大系列的初步数据，表明MALAT1可能在 盐敏感者和SS大鼠的肾脏，并可能影响盐敏感型高血压的发生 通过调节肾脏miR-214-3p/eNOS通路。我们建议调查MALAT1的S在 盐敏感型高血压的发生发展(目标1)，肾脏miR-214-3p/eNOS通路在该效应中的作用 MALAT1对高血压的作用(目标2)，以及潜在的分子相互作用(目标3)。我们将实现这些目标 目的通过对稀有的人体样本进行分析，在动物模型中进行组合基因操作， 以及包括基因组编辑和RafTOP(标记寡核苷酸提取的快速冷冻)在内的新方法。 RafTOP是我们最近开发的一种识别特定RNA的天然相互作用组的方法。

PROJECT SUMMARY More than 50% of hypertensive patients show an increased blood pressure sensitivity to salt intake. However, population-wide reduction of salt intake has proved to be difficult, making it ever more important to better understand the mechanism of salt-sensitive hypertension and provide a basis for developing new interventions. The kidney plays a key physiological role in the development of hypertension including salt- sensitive hypertension. Thousands of genes in the genome encode long non-coding RNAs (lncRNAs). lncRNAs can interact with and influence the function of other RNA or proteins. Few lncRNAs have been studied for their role in hypertension. Unlike most lncRNAs, MALAT1 (metastasis associated lung adenocarcinoma transcript 1; Malat1 in rodents) is conserved across many species and expressed at high abundance levels in several tissues including the kidney, which suggests MALAT1 might be important physiologically. However, MALAT1’s physiological and pathophysiological role remains largely unknown. We discovered recently that renal miR-214-3p targets and suppresses endothelial nitric oxide synthase (eNOS) directly, which contributes significantly to the development of salt-sensitive hypertension in rat models and possibly humans. This was supported by a systematic analysis of human sequence variants and all miRNA precursors, small RNA deep sequencing in human kidney biopsy specimens, kidney-specific inhibition of miR- 214-3p in Dahl SS rats, and a newly generated mutant rat strain. The Dahl SS rat is the model most widely used to study the molecular mechanism of human salt-sensitive hypertension. We have obtained a large series of preliminary data that suggest MALAT1 might be dysregulated in the kidneys of salt-sensitive humans and SS rats and might influence the development of salt-sensitive hypertension by regulating the renal miR-214-3p/eNOS pathway. We propose to investigate MALAT1’s role in the development of salt-sensitive hypertension (Aim 1), the role for the renal miR-214-3p/eNOS pathway in the effect of MALAT1 on hypertension (Aim 2), and the underlying molecular interactions (Aim 3). We will achieve these aims by using analysis of scarcely available human samples, combinatorial gene manipulation in animal models, and new methods including genome editing and RafTOP (rapid freezing with tagged oligonucleotide pullout). RafTOP is a method for identifying the native interactome for a specific RNA that we developed recently.