Mechanisms of Wg/Wnt regulation by glypican Dlp

磷脂酰肌醇蛋白聚糖 Dlp 调节 Wg/Wnt 的机制

• 批准号: 10371305
• 负责人: Indrayani Waghmare
• 金额: $ 10万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-01-01 至 2023-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10371305
• 关键词: Actins; Adhesions; Affect; Behavior; Binding; Biochemical; Cancer cell line; Carcinoma; Cell Adhesion; Cell Proliferation; Cell Shape; Cell surface; Cells; Complex; Data; Development; Distant; Drosophila genus; Ensure; Epithelial; Event; Extracellular Space; Genetic; Glypican; Growth; Homeostasis; Integral Membrane Protein; Investigation; Ligands; Link; Matrix Metalloproteinases; Mediating; Metalloproteases; Modeling; Molecular; Oogenesis; Organism; Output; Paracrine Communication; Pathway interactions; Pattern; Phase; Production; Proteins; Proteolysis; Proteomics; Regulation; Reporting; Role; Shapes; Signal Transduction; Source; Testing; Tissues; WNT Signaling Pathway; Wnt proteins; base; cell behavior; cell type; extracellular; fly; imaging approach; migration; novel; paracrine; tool; tumor; tumor growth; tumorigenesis; tumorigenic

PROJECT SUMMARY Wnt signaling is an evolutionarily conserved pathway that regulates several cellular behaviors such as cell proliferation, survival, differentiation, and migration to promote tissue homeostasis. Of note, the Wg/Wnt signaling pathway is important for tissue patterning and is often deregulated in epithelial cancers. Secreted Wnt ligands are distributed in the extracellular space to promote paracrine and long-range signaling in target cells. These paracrine and long-range functions of Wnts are dependent on extracellular Wnt availability, which in part is dictated by cell-surface glypican, Dally-like protein (Dlp). In this proposal, I will focus on molecular mechanisms that dictate Dlp-mediated regulation of Wnt availability and signaling. Dlp’s role in regulating Wnt signaling has been described as ‘biphasic’: By continual binding and release, Dlp simultaneously promotes long-range signaling and restricts paracrine signaling, ensuring proper ligand availability at both ranges. The Page-McCaw lab established Drosophila germarium, a tissue where oogenesis occurs, as a model to study mechanisms that define Wnt signaling ranges, and identified a novel Dlp/Mmp2 (Matrix Metalloprotease 2) module that modulates paracrine and long-range Wnt signaling in the germarium. Specifically, I found that proteolytic cleavage of Dlp by Mmp2 alters its subcellular localization and function to modulate Wnt availability. Additionally, my preliminary data suggest that Dlp/Mmp2 may regulate Wnt signaling in epithelial tumors to promote tumor growth. In Aim 1, I will investigate the molecular events that occur downstream of proteolytic cleavage of Dlp to modulate Wg/Wnt availability and signaling in the germarium and tumors. The extracellular Wnt distribution is tightly linked with its production and secretion. I found that Dlp can modulate Wg (Wnt-1) production in source cells in the germarium. Wg production in source cells in germaria is tightly regulated and this regulation is crucial for proper oogenesis. Additionally, I found that Dlp interacts with non-ligand proteins (a finding that has not been previously reported) that communicate with intracellular cytoskeletal machinery, potentially to modulate cell adhesion and/or shape. In Aim 2, I will investigate novel mechanisms of how Dlp regulates Wnt ligand production and long-range Wg distribution to facilitate long-range Wg signaling. These investigations will uncover previously unappreciated roles of cell-surface glypicans in regulating Wnt signaling and elucidate novel paradigms of developmental strategies employed in multicellular organisms to maintain tissue homeostasis.

项目摘要 Wnt信号传导是一种进化配置的途径，可调节几种蜂窝行为，例如 细胞增殖，生存，分化和迁移以促进组织稳态。值得注意的是，WG/Wnt 信号通路对于组织模式很重要，并且通常在上皮癌中受管制。分泌的Wnt 配体分布在细胞外空间中，以促进靶细胞中的旁分泌和远距离信号传导。 Wnt的这些旁分泌和远程功能取决于细胞外Wnt的可用性，部分部分 由细胞表面糖型，Dally样蛋白（DLP）决定。在此提案中，我将重点介绍分子机制 这决定了DLP介导的WNT可用性和信号传导的调节。 DLP在调节Wnt信号传导中的作用已被描述为“双相”：通过连续结合和释放， DLP只是促进远程信号传导并限制旁分泌信号传导，以确保正确的配体 两个范围的可用性。 Page-McCaw实验室建立了果蝇，这是一种卵子发生的组织 发生，作为研究定义Wnt信号范围并确定新型DLP/MMP2的模型 （基质金属蛋白酶2）调节寄生虫中旁分泌和远距离Wnt信号的模块。 具体而言，我发现MMP2对DLP的蛋白水解切割改变了其亚细胞定位和功能 调节可用性。此外，我的初步数据表明DLP/MMP2可能会调节Wnt信号传导 在上皮肿瘤中以促进肿瘤生长。在AIM 1中，我将研究发生的分子事件 DLP的蛋白水解裂解的下游，以调节胚芽中的WG/WNT可用性和信号传导 肿瘤。 细胞外Wnt分布与其生产和分泌紧密相关。我发现DLP可以 调节源细胞中源细胞的WG（WNT-1）产生。胚芽中源细胞中的WG产生是 严格调节，该调节对于适当的卵子发生至关重要。此外，我发现DLP与 非配体蛋白（以前尚未报道的发现）与细胞内通信 细胞骨架机械，可能调节细胞粘合剂和/或形状。在AIM 2中，我将调查小说 DLP如何调节Wnt配体产生和远程WG分布的机制以促进远程 WG信号传导。这些调查将发现以前没有批准的细胞表面果糖在 调节Wnt信号传导并阐明多细胞中使用的新型发展策略范式 维持组织稳态的生物体。