TGF-beta Pathways that Protect Epithelia in Chronic Renal Injury

慢性肾损伤中保护上皮细胞的 TGF-β 通路

• 批准号: 9294119
• 负责人: Leslie S Gewin
• 金额: $ 35.55万
• 依托单位: VANDERBILT UNIVERSITY MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-06-13 至 2020-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9294119
• 关键词: Acute Renal Failure with Renal Papillary Necrosis; Address; Adhesions; Affect; American; Angiotensins; Antioxidants; Apoptosis; Aristolochic Acids; Atrophic; Cell Culture Techniques; Cell Cycle; Cell Cycle Arrest; Cell Cycle Progression; Cell Differentiation process; Cell Survival; Cells; Chronic; Chronic Kidney Failure; Clinical Trials; Data; Economic Burden; Epithelial; Epithelial Cells; Epithelium; Etiology; Event; FDA approved; FOXO1A gene; Fibrosis; Functional disorder; G1 Arrest; Genetic; Genetic Models; Growth Factor; Impairment; In Vitro; Injury; Kidney; Ligand Binding; Light; Location; Mediating; Mediator of activation protein; Molecular Biology Techniques; Morbidity - disease rate; Mus; Organ; Pathway interactions; Pharmacology; Production; Proteins; Public Health; Role; Signal Pathway; Signal Transduction; Small Interfering RNA; Snails; Testing; Therapeutic; Transforming Growth Factor beta; Transforming Growth Factors; Tubular formation; base; biological adaptation to stress; cell type; genetic approach; improved; in vivo; inhibitor/antagonist; innovation; insight; migration; mortality; mouse model; novel; promoter; protective effect; public health relevance; receptor; renal epithelium; response; therapeutic target; tool; transcription factor

 DESCRIPTION (provided by applicant): Chronic kidney disease (CKD) is a growing public health problem that affects 10% of Americans, increases morbidity and mortality, and imposes a huge economic burden. Persistent renal tubular injury is an important component of tubulointerstitial fibrosis (TIF), the common feature of progressive CKD of any etiology. Growth factors such as transforming growth factor- (TGF) are important determinants of how tubula epithelia respond to injury. Although excessive TGF- signaling promotes TIF, blocking TGF-using genetic tools results in increased tubular damage after mouse models of CKD, suggesting that TGF-signaling has protective effects as well. TGF-mediates many differet cellular effects including G1 arrest of cell cycle progression and epithelial de-differentiation. Tis proposal tests the hypothesis that TGFsignaling in the proximal tubule promotes survival ater chronic renal injury by inducing cell cycle arrest and epithelial de- differentiation. The first ai investigates how TGF- alters cell cycle progression and how these changes affect epithelial survival using murine models of injury and pharmacologic inducers of G1 arrest. To test this, we will use genetically modified mice that lack the TGF receptor (TRII) specifically in the proximal tubule and induce chronic kidney injury by either angiotensin/ uninephrectomy or aristolochic acid. We anticipate that mice lacking TRII have impaired G1 arrest and reduced epithelial survival after injury and that this improves with administration of palbociclib, an FDA-approved inducer of G0/G1 cell cycle arrest. Proximal tubule cells with and without TRII in vitro will be used to understand the signaling pathways through which TGF- induces cell cycle arrest. The second aim explores mechanisms whereby TGFlters epithelial differentiation nd how these changes impact survival. The role of epithelial differentiation in chronic kidney injury will be defined using mice with epithelial-specific deletion of Snail, a key transcription factor downstream of TGF- Proximal tubule cells with and without TRII will be transfected with siRNA and constructs in vitro to test the hypothesis that Snail mediates TGF--dependent de-differentiation. The third aim defines how the Wnt/-catenin pathway, important in renal injury, interacts with TGF- to affect epithelial survival. Genetic models and cell culture techniques wil be used to test the hypothesis that TGF- increases canonical Wnt/catenin signaling to improve epithelial survival through increased de-differentiation and G0/G1 cell cycle arrest. This proposal uses innovative genetic approaches and a new strategy for analyzing injury in a quantitative and location-specific way. These studies will provide valuable insights into the roles of cell cycle and epithelial plasticity in the epithelial response to CKD, focusing on the proximal tubule which is the target of acute kidney injury and a mediator of CKD. However, information about cell cycle, epithelial plasticity and response to chronic injury has implications beyond the kidney as most organ dysfunction results from chronic epithelial injury.

 描述（由适用提供）：慢性肾脏疾病（CKD）是一个日益增长的公共卫生问题，影响了10％的美国人，增加了发病率和死亡率，并且不可能是巨大的经济伯恩。持续的肾结结核病是肾结龙纤维化（TIF）的重要组成部分，这是任何病因的进行性CKD的共同特征。生长因子（例如转化生长因子）（tgf）是小管上皮如何应对损伤的重要决定因素。尽管超过TGF-信号传导会促进TIF，但使用遗传工具阻止TGF-在CKD小鼠模型后会增加管状损伤，这表明TGF-signaling也具有保护效果。 TGF-降低了许多不同的细胞效应，包括G1停滞细胞周期进展和上皮脱不同。 TIS提案检验了代理管中TGF的假设通过诱导细胞周期停滞和上皮分化来促进生存的慢性肾脏损伤。第一个AI研究了TGF-如何使用损伤的鼠模型和G1停滞的药物诱导剂来改变细胞周期的进程以及这些变化如何影响上皮生存。为了测试这一点，我们将使用通常在近端管中缺乏TGF受体（TRII）的一般修饰的小鼠，并通过血管紧张素/非肾脏切除术或阿里斯托糖酸诱导慢性肾脏损伤。我们预计缺乏TRII的小鼠损害了G1停滞并降低了受伤后的上皮生存，并且随着FDA批准的G0/G1细胞周期停滞的诱导剂Palbociclib的给药而有所改善。带有和没有TRII的近端小管细胞将用于了解TGF-诱导细胞周期停滞的信号传导途径。第二个目的探索了TGF的机制。tgftresters上皮分化和这些变化如何影响生存。 The role of epithelial differentiation in chronic kidney injury will be defined using mice with epithelial-specific deletion of Snail, a key transcription factor downstream of TGF- Proximal tubele cells with and without TRII will be translated with siRNA and constructs in vitro to test the hypothesis that Snail mediates TGF--dependent de-differentiation.第三个目标定义了Wnt/-catenin途径在肾脏损伤中重要的如何与TGF-相互作用以影响上皮生存。遗传模型和细胞培养技术将用于检验以下假设：TGF-增加规范的Wnt/氨基信号传导，以提高脱差分和G0/G1细胞周期的停滞来提高上皮生存。该建议使用创新的遗传方法和新的策略，以定量和特定于位置的方式分析伤害。这些研究将为角色提供宝贵的见解 对CKD的上皮反应中的细胞周期和上皮可塑性的重点 Tubele是急性肾脏损伤的靶标和CKD介体。然而，由于慢性上皮损伤导致大多数器官功能障碍，因此有关细胞周期，上皮可塑性和对慢性损伤的反应的信息超出了肾脏。