DETERMINATION AND EXPRESSION OF AMELOGENIN GENE PRODUCTS

釉原蛋白基因产物的测定和表达

• 批准号: 7812613
• 负责人: Malcolm L. Snead
• 金额: $ 39.99万
• 依托单位: UNIVERSITY OF SOUTHERN CALIFORNIA
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-23 至 2013-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7812613
• 关键词: Address; Adipocytes; Adipose tissue; Bone Marrow; Cell Lineage; Cells; Cementoblast; Circadian Rhythms; Congenital Abnormality; Data; Defect; Dental Cementum; Dental Enamel; Employment; Enamel Formation; Fatty acid glycerol esters; Fibroblasts; Figs - dietary; Funding; Gene Expression; Genetic Transcription; Human; Human Genetics; In Vitro; Investigation; Jaw; Knockout Mice; Lead; Measures; Mediating; Mesenchymal Stem Cells; Metabolic Bone Diseases; Minerals; Monkeys; Mus; Muscle; National Institute of Dental and Craniofacial Research; Natural regeneration; Operative Surgical Procedures; Oral mucous membrane structure; Organism; Osteoblasts; Osteogenesis; Parents; Pathway interactions; Periodontal Ligament; Postdoctoral Fellow; Process; Protein Isoforms; Proteins; RNA Splicing; Rattus; Recovery; Regenerative Medicine; Regulation; Role; Scientist; Signal Pathway; Signal Transduction; Stem cells; Strategic Planning; Stromal Cells; Structural Protein; Structure; Testing; Therapeutic Intervention; Tissues; Tooth structure; Transcript; Transgenes; Transgenic Mice; Trauma; United States National Institutes of Health; Weaning; Wild Type Mouse; amelogenin; biomineralization; bone; bone loss; bone mass; bone sialoprotein; cell behavior; cell type; embryonic stem cell; factor C; improved; in vivo; inhibitor/antagonist; leucine-rich amelogenin peptide; lipid biosynthesis; novel; novel strategies; osteogenic; parent grant; public health relevance; research study; stem cell differentiation; transcription factor; tumor

DESCRIPTION (provided by applicant): This project is part of NOT-OD-09-058, with Notice Title, "NIH Announces the Availability of Recovery Act Funds for Competitive Revision Applications" and is consistent with the aims of parent grant DE06988 which seeks to determine if amelogenin proteins contribute to cell behavior by promoting signals that change differentiation. In this project we expand parent grant specific aim 3, to expand the scope of investigation into the role of amelogenin to include their capacity to induce an osteogenic pathway in stem cells. The scope of this Competitive Revision project will enhance the pace of discovery and it will provide additional employment for post-doctoral scientists. One of the amelogenin splicing isoforms, Leucine-rich Amelogenin Peptide (LRAP) has been shown to induce osteogenesis in various cell types, including rat muscle fibroblasts, mouse cementoblasts, mouse oral mucosal cells, and mouse embryonic stem cells. Our preliminary studies have shown that LRAP stimulates osteoblastogenesis and inhibits adipogenesis of bone marrow mesenchymal stem cells (BMMSC). Canonical Wnt/2-catenin signaling pathway is activated upon LRAP treatment; while a specific Wnt inhibitor sFRP-1, completely blocks LRAP-mediated lineage selection of BMMSCs. We hypothesize that LRAP is capable of modulating the reciprocal relationship between osteogenic and adipogenic cell lineages in BMMSCs through activating the canonical Wnt/2-catenin pathway. We propose the following specific aims to test this hypothesis. Aim I: To determine whether activation of the canonical Wnt/ 2 -catenin pathway by LRAP is necessary and sufficient to stimulate osteoblastogenesis and to inhibit adipogesis of BMMSCs in vitro. Aim II: To investigate the mechanism by which LRAP activates the canonical Wnt/2-catenin pathway and to characterize the functional domain(s) in LRAP. Aim III: To examine the in vivo effect of LRAP on bone and adipose tissue in transgenic mice expressing the LRAP transgene in bone marrow and in amelogenin null mice. PUBLIC HEALTH RELEVANCE: A previously unrecognized function of amelogenin proteins is to stimulate the differentiation of stem cells towards bone formation while suppressing their differentiation into fat forming cells. This switch is activated by amelogenin stimulating the Wnt signaling cascade. Utilizing amelogenin proteins may improve surgical correction of birth defects, trauma and tumor induced bone loss by providing a means to stimulate bone formation replacing foreign materials with real bone.

描述（由申请人提供）：该项目是NOT-OD-09-058的一部分，并带有通知标题：“ NIH宣布可用于竞争性修订应用程序的恢复ACT资金”，并且与父母Grant Grant De06988的目标一致，该目标旨在通过促进促进信号改变差异的信号来确定氨基蛋白蛋白是否对细胞行为有助于改变差异。在这个项目中，我们扩展了父授予特定的目标3，以扩大对氨基蛋白蛋白的作用的研究范围，以包括其在干细胞中诱导成骨途径的能力。这个竞争性修订项目的范围将提高发现的步伐，并为博士后科学家提供额外的就业机会。已证明，富含亮氨酸蛋白肽（LRAP）的蛋白蛋白剪接同工型之一，可诱导各种细胞类型的成骨，包括大鼠肌肉成纤维细胞，小鼠胶质细胞，小鼠口服粘膜粘膜细胞和小鼠胚胎干细胞。我们的初步研究表明，LRAP刺激成骨细胞生成并抑制骨髓间充质干细胞的脂肪形成（BMMSC）。 LRAP处理后，典型的Wnt/2-catenin信号通路被激活。而特定的Wnt抑制剂SFRP-1则完全阻止了LRAP介导的BMMSC谱系选择。我们假设LRAP能够通过激活规范的Wnt/2-catenin途径来调节BMMSC中成骨和成脂细胞谱系之间的相互关系。我们提出以下特定旨在检验该假设的特定旨在。目的I：确定通过LRAP激活规范的Wnt/ 2-蛋白蛋白途径是否需要且足以刺激成骨细胞生成并抑制体外BMMSCS的脂肪形成。 AIM II：研究LRAP激活规范Wnt/2-catenin途径的机制，并表征LRAP中功能域。 AIM III：检查LRAP对骨髓和氨基蛋白null小鼠中表达LRAP转基因的转基因小鼠中骨和脂肪组织的体内影响。 公共卫生相关性：先前无法识别的阿米他蛋白蛋白的功能是刺激干细胞向骨形成的分化，同时抑制其分化为脂肪形成细胞。通过氨基蛋白刺激Wnt信号级联反应来激活该开关。利用氨基蛋白蛋白可以通过提供一种刺激骨形成的手段来代替用真实骨头代替异物的手段来改善先天缺陷，创伤和肿瘤诱发骨质流失的手术纠正。