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(申请人提供)：角膜混浊是世界上第三大致盲原因，可能是由于内皮细胞营养不良、上皮营养不良、纤维化或由于伤口愈合失调导致基质混浊所致。角膜创面愈合在很大程度上受到细胞外环境的影响，既有可溶信号，也有生物物理信号，如基质形态和细胞外基质硬度。我们实验室和其他实验室的研究表明，细胞外基质的内在生物物理属性深刻地调节了细胞的一系列基本表型特征，如黏附、迁移、形状、大小和增殖，并伴随着激活/抑制特定的信号通路。尽管生物物理线索具有基本的重要性，但在我们对这些生物物理线索如何影响角膜基本细胞反应和伤口愈合的理解中，仍然存在着重大的知识缺口。在这项提案中，我们将确定介导角膜伤口愈合的信号通路的表达和定位影响。YAP(是相关蛋白)和TAZ(带有PDZ结合基序的转录共激活因子)作为一对转导分子连接细胞外环境的变化，如地形或基质硬度的变化。这两种蛋白质位于影响伤口愈合的多个信号通路的交叉点。初步数据表明，兔角膜基质的硬度在激光治疗性角膜切削术(PTK)后7天内增加了10倍。这与YAP/TAZ明显定位于上皮细胞的细胞核，导致其转录激活生长因子，如CTGF和TGFβ有关。在活体角膜损伤后，YAP/TAZ在前基质细胞中有较强的表达。这导致中心假说，YAP/TAZ的表达和定位是受外界生物物理刺激影响的角膜细胞信号事件的重要介质，是角膜创伤修复的基本部分。这一建议将描述YAP/TAZ在体外对角膜细胞的影响(A)，(B)在体内兔角膜伤口愈合中的影响，以及 (C)YAP、TAZ和体内双基因敲除条件基因敲除小鼠角膜伤口的愈合。这些研究将确定YAP/TAZ通过外部生物物理信号在上皮细胞中影响的信号通路，并将确定在伤口愈合过程中基质发生的变化。