Role of micro RNAs in renin cell differentiation

微小RNA在肾素细胞分化中的作用

• 批准号: 8443873
• 负责人: ROBERTO Ariel GOMEZ
• 金额: $ 36.29万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-04-01 至 2014-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8443873
• 关键词: Address; Adult; Affect; Arteries; Binding; Biological; Biological Assay; Blood Pressure; CREB1 gene; Cell Count; Cell Differentiation process; Cell Lineage; Cell model; Cells; Characteristics; Child; Cyclic AMP; Development; Electrolytes; Embryo; Embryonic Development; Enzymes; Epigenetic Process; Exclusion Criteria; Functional RNA; Gene Expression; Genes; Homeostasis; Hormones; Hypertension; In Vitro; Indium; Juxtaglomerular Cell; Kidney; Kidney Diseases; Label; Life; Liquid substance; Maintenance; Mediating; Medical; Mesenchyme; Methodology; MicroRNAs; Morphogenesis; Mus; Nephrons; Nucleotides; Partner in relationship; Pattern; Phenotype; Production; Regulation; Renin; Renin-Angiotensin System; Reporter; Role; Small RNA; Smooth Muscle; Smooth Muscle Myocytes; System; Testing; Transcriptional Regulation; Undifferentiated; Vascular Smooth Muscle; Vascularization; arteriole; blastema; cell motility; cell type; chromatin remodeling; fetal; gene repression; homologous recombination; human DICER1 protein; in vivo; interstitial cell; kidney vascular structure; mature animal; mesangial cell; migration; nephrogenesis; precursor cell; promoter; public health relevance; recombinase; response; screening

DESCRIPTION (provided by applicant): Renin, the key hormone of the renin-angiotensin system, regulates blood pressure and fluid-electrolyte homeostasis. In early embryonic development, renin precursor cells are present in the undifferentiated metanephric mesenchyme. Later in fetal life, renin cells are broadly distributed along intrarenal arteries and inside the glomeruli. With maturation, renin cells are fewer and restricted to the classical adult juxtaglomerular (JG) localization. If blood pressure or fluid-electrolyte homeostasis is threatened, the number of renin cells increases along preglomerular arteries, glomeruli and interstitium resembling the embryonic pattern. The increase in renin cell number (recruitment) is due to de-differentiation of preexisting adult cells and does not involve cell migration or replication. In fact, using a cre-lox system we found that renin cells are precursors that give rise to arteriolar smooth muscle (SM), mesangial, and interstitial cells and it is these cells that re-express renin when homeostasis is threatened. Further, using a dually labeled cell model we found that arteriolar SM cells reacquire the renin phenotype upon cAMP stimulation, an effect mediated by chromatin remodeling and binding of CREB at the cAMP responsive element in the renin promoter. In addition to this epigenetic/ transcriptional control, the acquisition and maintenance of renin cell identity may be regulated by endogenous microRNAs, a group of non-coding small RNAs that regulate gene expression at the post-transcriptional level and are known to regulate cell fate. Conditional deletion of Dicer in renin cells resulted in the disappearance of JG cells in the kidney suggesting that microRNAs may regulate renin cell specification and kidney vascular development. Using stringent exclusion criteria and a screening strategy involving multiple methodologies and functional assays, we identified two exciting and unique renin cell specific microRNAs (miR-330 and miR- 125b-5p) that regulate crucial genes for the maintenance of the myoepithelioid renin cell phenotype. We hypothesize that miR-330 and miR-125b-5p regulate the identity of the renin cell by controlling the expression of renin and vascular smooth muscle genes. Using in vivo and in vitro approaches, we will test the following hypotheses: 1) microRNAs determine the temporal and spatial pattern of renin cell differentiation during development, 2) miR-330 and miR-125b-5p have a characteristic distribution pattern and expression levels that regulate the state of differentiation of the renin cell, and 3) miR-330 and miR-125b-5p regulate the identity and fate of renin cells and nephron morphogenesis.

描述(申请人提供)：肾素，肾素-血管紧张素系统的关键激素，调节血压和水电解质平衡。在胚胎发育早期，肾素前体细胞存在于未分化的后肾间充质中。在胎儿生命后期，肾素细胞沿肾动脉和肾小球内广泛分布。随着肾素的成熟，肾素细胞减少，仅限于经典的成人肾小球旁(JG)定位。如果血压或水-电解质平衡受到威胁，肾素细胞的数量会沿着肾小球前动脉、肾小球和间质增加，类似于胚胎模式。肾素细胞数量的增加(募集)是由于先前存在的成体细胞去分化所致，与细胞迁移或复制无关。事实上，使用cre-lox系统，我们发现肾素细胞是产生小动脉平滑肌(SM)、系膜细胞和间质细胞的前体细胞，当动态平衡受到威胁时，正是这些细胞重新表达肾素。此外，使用双标记细胞模型，我们发现小动脉SM细胞在cAMP刺激下重新获得肾素表型，这一效应是由染色质重塑和CREB与肾素启动子中cAMP反应元件的结合所介导的。除了这种表观遗传/转录调控外，肾素细胞特性的获得和维持可能受到内源性microRNAs的调控，内源性microRNAs是一组非编码的小RNA，在转录后水平调节基因表达，并已知调节细胞命运。肾素细胞条件缺失导致肾脏JG细胞消失，提示microRNAs可能调节肾素细胞规格和肾脏血管发育。使用严格的排除标准和涉及多种方法和功能分析的筛选策略，我们鉴定了两个令人兴奋的独特的肾素细胞特异性microRNAs(miR-330和miR-125b-5p)，它们调节维持肌上皮样肾素细胞表型的关键基因。我们假设miR-330和miR-125b-5p通过控制肾素和血管平滑肌基因的表达来调节肾素细胞的特性。我们将利用体内和体外的方法检验以下假设：1)microRNAs决定肾素细胞发育过程中分化的时空模式；2)miR-330和miR-125b-5p具有调节肾素细胞分化状态的特有的分布模式和表达水平；3)miR-330和miR-125b-5p调控肾素细胞和肾单位形态发生的身份和命运。