Modulation of Signal Transduction by Nano-Topography

纳米形貌调制信号转导

• 批准号: 8806561
• 负责人: CHRISTOPHER John MURPHY
• 金额: $ 38.26万
• 依托单位: UNIVERSITY OF CALIFORNIA AT DAVIS
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-09-01 至 2018-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8806561
• 关键词: Accidents; Adhesions; Affect; Anterior; Binding; Blindness; Cell Adhesion; Cell Nucleus; Cell Proliferation; Cell Shape; Cells; Characteristics; Chronic; Cicatrix; Clinical Management; Complex; Contact Lenses; Cornea; Corneal Diseases; Corneal Injury; Corneal Opacity; Corneal Stroma; Cues; Data; Deposition; Doxycycline; Environment; Epithelial; Epithelial Cells; Epithelium; Event; Extracellular Matrix; Eye; Fibroblasts; Fibrosis; Gene Expression; Genes; Genetic Transcription; Goals; Growth Factor; Healed; Health; Homeostasis; In Vitro; Knock-out; Knockout Mice; Knowledge; Laboratories; Lead; Link; Maintenance; Mechanics; Mediating; Mediator of activation protein; Modeling; Molecular; Mus; Myofibroblast; Nanotopography; Operative Surgical Procedures; Oryctolagus cuniculus; Outcome; Participant; Play; Process; Property; Proteins; Reporting; Research; Role; Shapes; Signal Pathway; Signal Transduction; Stimulus; Stratification; Stromal Cells; Therapeutic Agents; Tissues; Transcription Coactivator; Transcriptional Activation; Transducers; Ulcer; Vision; Wound Healing; cell behavior; cell motility; cellular transduction; clinically relevant; connective tissue growth factor; corneal epithelium; corneal repair; differential expression; extracellular; healing; improved; in vivo; migration; nanoscale; new therapeutic target; novel; physical property; protein expression; research study; response

DESCRIPTION (provided by applicant): Corneal opacities, the third major cause of blindness in the world, may be a result of endothelial dystrophies, epithelial dystrophies, fibrosis or a haz in the stroma due to dysregulated wound healing. Corneal wound healing is largely influenced by the extracellular environment both through soluble signals and from biophysical cues such as substratum topography and extracellular matrix stiffness. Research from our lab and others have demonstrated intrinsic biophysical attributes of the extracellular matrix profoundly modulate a host of fundamental phenotypic characteristics of cells such as adhesion, migration, shape, size and proliferation accompanied by activation/inhibition specific signaling pathways. Despite the fundamental importance of biophysical cues, a significant knowledge gap remains in our understanding of how these biophysical cues influence basic cellular responses and wound healing in the cornea. In this proposal we will determine the expression and localization impact of signaling pathways that mediate corneal wound healing. YAP (Yes- associated protein) and TAZ (transcriptional co-activator with PDZ-binding motif) serve as a pair of transducers linking changes in the extracellular environment, such as a change in topography or of matrix stiffness. These two proteins are at the intersection of multiple signaling pathways that influence wound healing. Preliminary data demonstrate that the stiffness of the corneal stroma increases by 10-fold within 7 days after wounding by phototherapeutic keratectomy (PTK) in rabbits. This was associated with YAP/TAZ being markedly localized in the nucleus of epithelial cells resulting in their transcriptional activation of growth factors such as CTGF and TGF¿. A robust expression of YAP/TAZ was observed in the anterior stromal cells in vivo following corneal wounding. This leads to the central hypothesis that expression and localization of YAP/TAZ are essential mediators of signaling events in corneal cells influenced by external biophysical stimuli and are a fundamental part of corneal wound repair. This proposal would delineate the influence of YAP/TAZ in corneal cells (A) in vitro, (B) in the healing of corneal wounds in rabbits in vivo, and (C) the healing of corneal wounds in conditional knockout mice for YAP, TAZ and the double knockout in vivo. These studies will define the signaling pathways influenced by YAP/TAZ mediated by external biophysical cues in the epithelial cells and will also identify changes that occur in the stroma during wound healing.

描述（由适用提供）：角膜骨是世界上失明的第三个主要原因，可能是内皮营养不良，上皮营养不良，纤维化，纤维化或由于伤口失调导致的基质中的HAZ的结果。角膜伤口愈合在很大程度上受细胞外环境的影响，无论是通过固体信号和生物物理提示，例如底层地形和细胞外基质刚度。我们实验室和其他人的研究表明，细胞外基质的内在生物物理特性深刻地调节了细胞的基本表型特征，例如广告，迁移，形状，形状，大小和通过激活/抑制特异性信号通路完成。尽管生物物理提示具有根本的重要性，但我们对这些生物物理提示如何影响角膜中基本细胞反应和伤口愈合的理解仍然存在着重要的知识差距。在此提案中，我们将确定中位角膜伤口愈合的信号传导途径的表达和定位影响。 YAP（与蛋白质相关的蛋白质）和TAZ（具有PDZ结合基序的转化共激活因子）用作连接细胞外环境变化的一对换能器，例如地形的变化或基质刚度的变化。这两种蛋白质处于影响伤口愈合的多个信号通路的交点。初步数据表明，角膜基质的刚度在兔子中光疗法角膜切除术（PTK）绕组后的7天内增加了10倍。这与YAP/TAZ明显定位在上皮细胞的核中，导致其转录激活生长因子（例如CTGF和TGF。在角膜赢得后体内在体内的前基质细胞中观察到了YAP/TAZ的强大表达。这导致了一个中心假设，即YAP/TAZ的表达和定位是受外部生物物理刺激影响的角膜细胞中信号事件的重要介体，并且是角膜伤口修复的基本部分。该建议将描述YAP/TAZ在角膜细胞中的影响（a）体外，（b）体内兔子角膜伤口的愈合，以及 （c）有条件的敲除小鼠的角膜伤口在体内的YAP，TAZ和双基因敲除。这些研究将定义由上皮细胞中外部生物物理提示介导的YAP/TAZ影响的信号传导途径，还将确定伤口愈合过程中基质中发生的变化。