Tyrosine Phosphorylation in Lens Cell Differentiation

晶状体细胞分化中的酪氨酸磷酸化

• 批准号: 7879262
• 负责人: A. Sue Menko
• 金额: $ 38.24万
• 依托单位: THOMAS JEFFERSON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-06-02 至 2012-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7879262
• 关键词: Adherens Junction; Blindness; Cadherins; Cataract; Cataract Extraction; Cell Differentiation process; Cell Polarity; Cell Proliferation; Cell Survival; Cell physiology; Cell-Cell Adhesion; Cells; Complex; Cytoskeleton; Desmosomes; Detergents; Development; Epithelial Cells; Epithelium; Filament; Goals; Image; Intercellular Junctions; Intermediate Filaments; Investigation; Lens Diseases; Lens Fiber; Lens Opacities; Link; MAPK14 gene; Maintenance; Methods; Microfilaments; Modeling; Molecular; Morphogenesis; N-Cadherin; Pathway interactions; Pharmacologic Substance; Phenotype; Phosphotransferases; Prevention; Property; Protein Tyrosine Phosphatase; Proteins; Reactive Oxygen Species; Recruitment Activity; Regulation; Retina; Role; Signal Pathway; Signal Transduction; Small Interfering RNA; Stress; Stress Fibers; Structure; Structure-Activity Relationship; Tyrosine Phosphorylation; Undifferentiated; desmoplakin; epithelial to mesenchymal transition; fiber cell; lens; novel; prevent; public health relevance; src-Family Kinases

DESCRIPTION (provided by applicant): Cadherin cell-cell adhesion junctions establish the epithelial cell phenotype, prevent their epithelial to mesenchymal transition, define cell cytoarchitecture and signal morphogenesis. We seek to understand the function of these cell-cell junctions as guardians against the induction of cataract. Our studies show a unique structure/function relationship between cadherin junctions, their cytoskeletal linkages and lens differentiation state. Dynamic, highly detergent-soluble N-cadherin/2-catenin adherens junctions define the cytoarchitecture of lens epithelial cells while establishment of the highly ordered and elongated cytoarchitecture of differentiating lens fiber cells involves stabilization of cadherin junctions. We discovered the presence of a novel N-cadherin junctional complex in lens fiber cells, linked to the intermediate filament cytoskeleton through 3-catenin. We will investigate this novel cadherin complex for its role in providing the elongating lens fiber cells with the unique structural properties required for their differentiation and stability. In addition, we will study the differentiation-specific role of N-cadherin junctional complexes in the organization of the cytoskeleton and examine the role of the cytoskeleton in stabilizing cadherin differentiation-specific function. Interestingly, Src family tyrosine kinases (SFKs), regulators of cadherin junctions, have a dual role in lens differentiation. The SFK Src maintains lens epithelial cells in a proliferative, undifferentiated state and the SFK Fyn signals lens fiber cell differentiation. In this proposal we continue our investigation of the mechanisms of SFK regulation of N-cadherin junctional stability, and N-cadherin-linked SFK signaling of lens cell differentiation, in a pathway involving PI3K and Rac. Our previous studies show that the unregulated, constitutive activation of Src kinases disassembles lens cadherin junctions. We now find in a culture model for stress-induced cataract that lens opacities result from activation of a p38 stress kinase/Src kinase signaling pathway that targets the cadherin junctions. If Src kinase activation is blocked, both cadherin junctions and normal lens cytoarchitecture are maintained, and the development of lens opacities is prevented. We will investigate the mechanism of N-cadherin junction destabilization in cataract, focusing on the role protein tyrosine phosphatases (PTPs) as molecular regulators of both Src kinases and cadherin junctions. PTPs are inactivated by reactive oxygen species, a common cause of stress-induced cataract. It is hoped that in the long-term, understanding of this pathway will provide suitable pharmaceutical targets for the prevention of cataract. Will investigate our hypotheses in three specific aims: (1) Examine the hypothesis that N-cadherin regulates the cytoskeleton to establish and maintain lens structure and stability; (2) Examine the hypothesis that the Src kinases regulate lens cell differentiation state by their action as both upstream regulators and downstream signaling effectors of cadherin junctions; (3) Examine the hypothesis that misregulation of Src kinases abrogates the role of cadherin junctions and the cytoskeleton as protectors of lens structural stability and guardians against cataract. PUBLIC HEALTH RELEVANCE: Cataracts are opacities of the lens that interfere with its ability to focus an image on the retina. While many of the properties of cataract have been determined, the mechanism of cataract induction and potential methods of cataract prevention still remain elusive. The long term goals of this proposal are to understand the role of cadherin cell-cell junctions as guardians against this lens disease.

描述（由申请方提供）：钙粘蛋白细胞-细胞粘附连接建立上皮细胞表型，阻止其上皮向间充质转化，定义细胞结构和信号形态发生。我们试图了解这些细胞连接的功能，作为监护人对白内障的诱导。我们的研究表明，钙粘蛋白连接，它们的细胞骨架连接和透镜分化状态之间的独特的结构/功能关系。动态的，高度去污剂可溶的N-钙粘蛋白/2-连环蛋白粘附连接定义了透镜上皮细胞的细胞结构，而分化中的透镜纤维细胞的高度有序和细长的细胞结构的建立涉及钙粘蛋白连接的稳定。我们在透镜纤维细胞中发现了一种新的N-钙粘蛋白连接复合物，通过3-连环蛋白与中间丝细胞骨架相连。我们将研究这种新的钙粘蛋白复合物的作用，提供了独特的结构特性所需的分化和稳定的伸长透镜纤维细胞。此外，我们将研究分化特异性作用的N-钙粘蛋白连接复合物的组织中的细胞骨架和检查的作用，稳定钙粘蛋白分化特异性功能的细胞骨架。有趣的是，Src家族酪氨酸激酶（SFKs），钙粘蛋白连接的调节剂，在透镜分化中具有双重作用。SFK Src维持透镜上皮细胞处于增殖、未分化状态，SFK Fyn发出透镜纤维细胞分化的信号。在这个建议中，我们继续我们的调查机制的SFK调节N-钙粘蛋白连接的稳定性，和N-钙粘蛋白连接的SFK信号的透镜细胞分化，在一个途径，涉及PI 3 K和Rac。我们以前的研究表明，Src激酶的不受调节的组成性激活会分解透镜钙粘蛋白连接。我们现在发现，在应激诱导的白内障的培养模型中，透镜混浊是由靶向钙粘蛋白连接的p38应激激酶/Src激酶信号通路的激活引起的。如果Src激酶激活被阻断，钙粘蛋白连接和正常的透镜细胞结构得以维持，透镜混浊的发展得以防止。我们将研究白内障中N-钙粘蛋白连接不稳定的机制，重点是蛋白酪氨酸磷酸酶（PTPs）作为Src激酶和钙粘蛋白连接的分子调节剂的作用。PTPs被活性氧灭活，这是应激性白内障的常见原因。希望从长远来看，对这一途径的了解将为预防白内障提供合适的药物靶点。我们将从三个方面来研究我们的假说：（1）验证N-钙粘蛋白调节细胞骨架以建立和维持透镜结构和稳定性的假说：（2）验证Src激酶通过作为钙粘蛋白连接的上游调节因子和下游信号效应因子来调节透镜细胞分化状态的假说;（3）检验Src激酶的失调消除了钙粘蛋白连接和细胞骨架作为透镜结构稳定性的保护者和防止白内障的监护者的作用的假设。公共卫生相关性：白内障是透镜的混浊，干扰其在视网膜上聚焦图像的能力。虽然白内障的许多性质已经确定，但白内障诱导的机制和预防白内障的潜在方法仍然难以捉摸。这项建议的长期目标是了解钙粘蛋白细胞-细胞连接作为对抗这种透镜疾病的监护人的作用。