Regulatory RNA in Hypertension

项目摘要 超过50%的高血压患者表现出血压对盐摄入的敏感性增加。 然而，在全人口范围内减少盐的摄入量已被证明是困难的，因此， 进一步了解盐敏感性高血压的发病机制，为开发新的 干预措施。肾脏在高血压的发展中起着关键的生理作用，包括盐- 敏感性高血压 基因组中的数千个基因编码长非编码RNA（lncRNA）。lncRNA可以相互作用 影响其他RNA或蛋白质的功能。很少有lncRNA被研究它们在以下方面的作用： 高血压与大多数lncRNA不同，MALAT 1（转移相关肺腺癌转录物1; Malat 1 在啮齿动物中）在许多物种中是保守的，并在几种组织中以高丰度水平表达 包括肾脏，这表明MALAT 1可能在生理上很重要。然而，MALAT 1的 生理和病理生理作用仍然在很大程度上未知。 我们最近发现肾脏miR-214- 3 p靶向并抑制内皮型一氧化氮合酶， 在盐敏感性高血压大鼠模型中，内皮型一氧化氮合酶（eNOS）直接参与盐敏感性高血压的形成 也可能是人类这得到了人类序列变异和所有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- 3 p的Dahl SS大鼠，和一个新产生的突变大鼠品系。Dahl SS大鼠是最广泛使用的模型 探讨人类盐敏感性高血压的分子机制。 我们已经获得了大量的初步数据，表明MALAT 1可能在人乳腺癌中表达失调。 盐敏感的人和SS大鼠的肾脏，并可能影响盐敏感性高血压的发展 通过调节肾脏miR-214- 3 p/eNOS通路。我们建议研究MALAT 1在 盐敏感性高血压的发展（目的1），肾脏miR-214- 3 p/eNOS通路在影响 MALAT 1对高血压的作用（目的2），以及潜在的分子相互作用（目的3）。我们将实现这些目标 目的是通过分析几乎不可用的人类样本，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（快速冷冻与标记寡核苷酸拉出）的新方法。 RafTOP是我们最近开发的一种用于识别特定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.