Structure/Function Relationship of the Lumican Gene

Lumican基因的结构/功能关系

• 批准号: 9096796
• 负责人: WINSTON W KAO
• 金额: $ 54.43万
• 依托单位: UNIVERSITY OF CINCINNATI
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-04-01 至 2018-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9096796
• 关键词: Accounting; Adenoviruses; Amino Acids; Apoptosis; Binding; Biochemical; Biological; Biological Assay; C-terminal; Cell Cycle; Cell Differentiation process; Cell Line; Cell Proliferation; Cells; Chimeric Proteins; Complex; Computer Simulation; Cornea; Corneal Injury; Corneal Stroma; Cytoskeleton; Data; Debridement; Defect; Development; Diabetic mouse; Disease; Down-Regulation; Doxycycline; Embryonic Development; Epidermal Growth Factor Receptor; Epiregulin; Epithelial Cells; Epithelium; Extracellular Matrix; Family; Genes; Growth; Healed; Health; Hela Cells; Homeostasis; Human; Immunofluorescence Immunologic; Impaired wound healing; In Vitro; Injury; Keratan Sulfate; Knock-out; Lead; Length; Leucine; Lifting; Ligands; MAPK3 gene; Maintenance; Mediating; Messenger RNA; Modeling; Molecular; Mus; Nuclear Translocation; Pathogenesis; Peptides; Phosphorylation; Phosphotransferases; Polymers; Production; Proteins; Proteoglycan; Recombinants; Role; Signal Transduction; Signaling Molecule; Staining method; Stains; Structure-Activity Relationship; TGF-beta type I receptor; TGFBR1 gene; TGFBR2 gene; Telomerase; Testing; Therapeutic; Tissues; Transforming Growth Factor beta; Transgenic Mice; Up-Regulation; Wound Healing; cell motility; corneal epithelium; diabetic; diabetic patient; efficacy testing; healing; in vivo; knock-down; lumican; molecular dynamics; mouse model; mutant; novel; peptide drug; polymerization; prevent; receptor; response

DESCRIPTION (provided by applicant): Lumican (Lum) belongs to the small leucine rich proteoglycan family and is a constituent of the extracellular matrix and serves as a matrikine (a term pertaining to ECM components and their proteolytic peptides that are able to regulate cell activity). Lum contributes to corneal transparency and is essential for corneal wound healing, but little is known regarding how this is achieved. Our recent findings suggest that TGFß type I receptor (TGFBR1/ALK5) is a novel Lum receptor. We aim to examine the biological significance of Lum/ALK5 interaction accounting for the pleiotropic function of Lum and TGF� signaling in regulating various cellular responses during embryonic development and pathogenesis. Lumican binds ALK5 and promotes wound healing of HTCE (human telomerase-immortalized corneal epithelial) cells by alleviating the suppression of cell proliferation and up-regulating EGFR-ligands to sustain ERK activation. Specific Aim 1 will examine the mechanism responsible for the enhanced in vitro wound healing elicited by the Lum/ALK5 complex. This will be achieved by examining the fate of TGFBR polymers following Lum binding to ALK5, by assessing the role of epiregulin up-regulation and through the identification of the ALK5 domain(s) required for interacting with Lum and or Src. Specific Aim 2 will examine the in vivo mechanism by which Lum/ALK5 promotes wound healing. This will be achieved through the use of various transgenic mouse models to examine the hypothesis that the ALK5/TGFBR2 polymer is required for Lum binding and subsequent promotion of wound healing. In addition this aim, will further examine the ALK5 intracellular domain that may mediate wound healing. Finally Specific Aim 3 will set out to identify and characterize therapeutic LumC peptides for wound healing, specifically the compromised wound healing seen in diabetic patients. The completion of the proposed studies will not only expand our understanding beyond the current central dogma of signaling cascades elicited by TGFß, but will also identify and characterize therapeutic peptides for treating persistent epithelium defects.

描述（由申请人提供）：Lumican（Lum）属于富含小亮氨酸的蛋白聚糖家族，是细胞外基质的组成成分，并作为基质因子（与ECM组分及其能够调节细胞活性的蛋白水解肽有关的术语）。Lum有助于角膜透明度，对角膜伤口愈合至关重要，但关于如何实现这一点知之甚少。我们最近的研究结果表明，TGF β 1型受体（TGFBR 1/ALK 5）是一种新的Lum受体。我们的目的是研究Lum/ALK 5相互作用的生物学意义，解释Lum和TGF β信号在胚胎发育和发病过程中调节各种细胞反应的多效性功能。Lumican结合ALK 5并通过减轻细胞增殖的抑制和上调EGFR配体以维持ERK活化来促进HTCE（人端粒酶永生化角膜上皮）细胞的伤口愈合。具体目标1将检查负责由Lum/ALK 5复合物引起的体外伤口愈合增强的机制。这将通过检查Lum与ALK 5结合后TGFBR聚合物的命运、通过评估表皮调节蛋白上调的作用以及通过鉴定与Lum和/或Src相互作用所需的ALK 5结构域来实现。具体目标2将检查Lum/ALK 5促进伤口愈合的体内机制。这将通过使用各种转基因小鼠模型来检查ALK 5/TGFBR 2聚合物是Lum结合和随后促进伤口愈合所需的假设来实现。此外，这一目标将进一步研究可能介导伤口愈合的ALK 5细胞内结构域。最后，具体目标3将着手鉴定和表征用于伤口愈合的治疗性LumC肽，特别是在糖尿病患者中观察到的受损伤口愈合。所提出的研究的完成将不仅扩展我们的理解，超越目前的中心法则的信号级联引起的TGF β，但也将确定和表征治疗肽治疗持续性上皮缺陷。