TGF-beta signaling in the kidney

描述（由申请人提供）：转化生长因子β 1（TGF-β 1）是一种多效性细胞因子，其控制多种细胞功能，包括细胞增殖、分化、凋亡和细胞外基质（ECM）合成。TGF-β 1是ECM蛋白合成和积累的有效诱导剂，并且作为包括肾脏在内的多种组织中纤维化发生的中心介质在进行性疾病的发病机制中起关键作用。TGF-β 1的作用是通过TGF-β I型（TssRI）和II型（TssRII）受体介导的，以激活细胞内途径。许多中心问题仍然与不同的受体如何以细胞特异性和环境特异性的方式介导TGF-β 1信号以引起多种细胞反应有关。我们研究的总体目标是了解肾损伤和纤维化的细胞和分子机制。主要目的是研究TGF-β 1信号通路的机制及其在肾脏损伤反应中的调节和功能作用。我们的假设是，自噬是一种适应性应激反应，通过抑制细胞凋亡和促进肾细胞存活来保护肾损伤，TGF-β 1通过调节自噬发挥细胞保护作用。此外，我们假设通过TAK 1-MKK 3-p38的TGF β 1信号传导是组织损伤反应的关键介质，其中TGF β 1调节自噬蛋白，这反过来又防止细胞凋亡和过度ECM积累。本研究将重点研究TAK 1介导的TGF-β 1信号通路在肾细胞中的作用机制和功能，以及在肾纤维化实验模型中对自噬的调节及其生理功能作用。具体目标是：具体目标1：目的2：研究TGF-β 1介导的自噬在肾细胞中的调控作用。具体目标3：确定自噬在肾纤维化实验模型中的体内生理功能作用。我们将采用最先进的方法，包括信号转导分子的各种显性失活突变体，MAPKs，专注于TAK 1-MKK 3信号传导轴，通过使用短干扰RNA（siRNA）进行基因沉默，以及遗传改变的小鼠，各种TAK 1，MKK 3，Caveolin-1和自噬基因LC 3和Beclin 1的无效小鼠。相关性：虽然TGF-β 1在肾纤维化发展中的中心作用已被充分证实，但不加区别地抑制TGF-β 1作用的一般策略可能被证明是轻率的。本研究将为进一步了解TGF-β 1信号转导的分子机制提供重要和新颖的信息，我们可能能够选择性地阻断TGF-β 1有害作用的信号通路。

DESCRIPTION (provided by applicant): Transforming growth factor-beta 1 (TGF-ss1) is a pleiotropic cytokine which controls multiple cellular functions including cell proliferation, differentiation, apoptosis, and extracellular matrix (ECM) synthesis. TGF-ss1 is a potent inducer of ECM protein synthesis and accumulation, and plays a key role in the pathogenesis of progressive diseases as a central mediator of fibrogenesis in a variety of tissues, including the kidney. TGF-ss1 actions are mediated via TGF-ss type I (TssRI) and type II (TssRII) receptors to activate intracellular pathways. Many central questions remain relating to how the distinct receptors mediate TGF-ss1 signals in a cell-specific and context-specific manner to elicit multiple cellular responses. The overall objective of our research is to understand the cellular and molecular mechanisms of renal injury and fibrosis. The major goals are to investigate the mechanisms of TGF-ss1 signaling pathways and their regulation and functional role in injury responses in the kidney. Our hypothesis is that autophagy represents an adaptive stress response to protect against renal injury by inhibiting apoptosis and promoting renal cell survival, and that TGF-ss1 exerts cytoprotective effects via regulating autophagy. Furthermore, we hypothesize that TGF-ss1 signaling via TAK1- MKK3-p38 is the critical mediator of tissue injury response in which TGF-ss1 regulates autophagy proteins which in turn prevents apoptosis and excessive ECM accumulation. This proposal will focus on examining the mechanism and functional role of TGF-ss1 signaling via TAK1 in renal cells, and the regulation of autophagy and its physiological functional role in an experimental model of renal fibrosis. The Specific Aims are: Specific Aim 1: To determine the mechanism and functional role of TGF-ss1 signaling via TAK1 in renal cells Specific Aim 2: To determine the regulation and function of autophagy induced by TGF-ss1 in renal cells. Specific Aim 3: To determine the in vivo physiological functional role of autophagy in an experimental model of renal fibrosis. We will employ state-of-the art approaches including a variety of dominant negative mutants of the signal transducing molecules, the MAPKs, focusing on the TAK1-MKK3 signaling axis, gene silencing by the use short interfering RNA (siRNA), and genetically altered mice, the null mice for the various TAK1, MKK3, Caveolin-1, and the autophagy genes, LC3 and Beclin 1. Relevance: Although the central role of TGF-ss1 in the development of renal fibrosis is well documented, general strategies to indiscriminately inhibit TGF-ss1 actions altogether may prove to be imprudent. The studies in this proposal will yield important and novel information in furthering our understanding of the molecular mechanisms of TGF-ss1 signal transduction, that we may be able to selectively block the pathway that signals the deleterious effects of TGF-ss1.