BMPs in Skeletal Growth and Osteogenic Differentiation

BMP 在骨骼生长和成骨分化中的作用

• 批准号: 7588763
• 负责人: Karen M. Lyons
• 金额: $ 31.36万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-04-01 至 2011-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7588763
• 关键词: Accounting; Achondroplasia; Address; Authorization documentation; BMPR1A gene; Biological Assay; Bone Morphogenetic Proteins; California; Cartilage; Cell Line; Cells; Chondrocytes; Chondrogenesis; Data; Defect; Development; Disclosure; Elements; Epiphysial cartilage; Equilibrium; Erinaceidae; Event; Exhibits; FGFR3 gene; Face; Fibroblast Growth Factor; Funding; Future; Gene Expression; Generations; Genetic screening method; Goals; Growth; Human Resources; In Vitro; Individual; Instruction; Laboratory Research; Last Name; Left; Limb structure; Los Angeles; Maintenance; Mediating; Mediator of activation protein; Modeling; Mus; Mutation; Names; Nuclear Translocation; Orthopedic Surgery procedures; Output; Pathway interactions; Phenotype; Phosphorylation; Physical condensation; Play; Postdoctoral Fellow; Pregnancy; Principal Investigator; Printing; Registries; Regulation; Relative (related person); Reporter; Research Personnel; Research Project Grants; Resolution; Role; Signal Pathway; Signal Transduction; Skeleton; Staging; Testing; Thanatophoric Dysplasia; Time; Tissue Engineering; Transducers; Transgenes; Universities; Washington; bone morphogenetic protein receptors; chondrodysplasia; craniofacial; human embryonic stem cell; in vivo; medical schools; mutant; osteogenic; osteogenin; prevent; programs; receptor; receptor function; repaired; response; skeletal; skeletogenesis; spine bone structure

This proposal investigates the roles of bone morphogenetic proteins (BMPs) in chondrogenesis. Through analysis of mice lacking BMPR1A and/or BMPR1B, we demonstrated that signaling through these receptors is essential for chondrocyte proliferation, differentiation, and maintenance. These studies showed that BMPR1A and BMPR1B have overlapping functions at the condensation stage, but did not reveal the extent to which they collaborate at later stages. Moreover, while ActRI is insufficient on its own to support chondro- genesis beyond the condensation stage, the extent to which BMPR1A and/or BMPR1B alone can promote chondrogenesis is unknown. Resolution of this issue is important for future strategies targeted toward activation of specific BMP receptors in order to elicit defined responses in tissue engineering applications or in repair in vivo. We address these issues in Aim 1through characterization ActRI/Bmprla and ActRI/Bmprlb double mutants, and through the generation of mice in which loss of BMP receptor function can be induced at late gestation stages. It is likely that BMPs transduce the majority of their effects through Smads, but it is unknown whether or not all 3 BMP-specific Smads are required for chondrogenesis, or whether Smads have distinct vs. overlapping functions. We address this issue in aim two. Studies of Bmpr1a/1b compound mutants uncovered evidence that loss of BMP signaling is accompanied by increased output from FGF pathways. This finding suggests that BMP and FGF pathways act antagonistically during chondrogenesis, and raises the possibility that the relative balance between BMP and FGF signaling is a major determinant of chondrocyte progression through the growth plate. We test this and identify downstream components of FGF signaling pathways responsible for these effects in aim three through construction of mice in which relative output from BMP and FGF pathways is altered, and through the use of limb cultures. Analysis of BMP receptor mutants demonstrated that BMP pathways are essential for expression of Ihh, the key regulator of chondrocyte proliferation and differentiation in the growth plate. It has been shown in vitro that ERK1/2 antagonizes BMP signaling by directly phosphorylating, and thereby inactivating, BMP-specific Smads. In aim four, we test whether this mechanism is relevant to Ihh expression in the growth plate. Understanding how BMPs control distinct aspects of chondrogenesis and how FGFs antagonize these effects is vital to understanding how BMPs can be utilized most optimally in tissue engineering applications, for cartilage repair and maintenance, and potentially for treatment of chondrodysplasias caused by activating FGF mutations. University of California, Los Angeles David Geffen School of Medicine Department of Orthopaedic Surgery 2614 MacDonald Research Laboratories 675 Charles E Young Dr. South Los Angeles, CA 90095 PHS 398 (Rev.09/04) Page 2 Form Page 2 Principal Investigator/Program Director (Last, First, Middle): KEY PERSONNEL. See instructions. Use continuation pages as needed to provide the required Start with Principal Investigator. List all other key personnel in alphabetical order, last name first. Name eRA Commons User Name Organization Karen Lyons LYONS2 UCLA Sean Brugger UCLA OTHER SIGNIFICANT CONTRIBUTORS Name Organization Akiko Hata Tufts University David Ornitz Washington University Lyons, Karen M information in the format shown below. Role on Project PI Postdoctoral fellow Role on Project Consultant Consultant Human Embryonic Stem Cells E3 No D Yes If the proposed project involves human embryonic stem cells, list below the registration number of the specific cell llne(s) from the following list: http://stemcells.nih.qov/reqistrv/index.asp. Usecontinuation pages as needed. If a specific line cannot be referenced at this time, include a statement that one from the Registry will be used. Cell Line Disclosure Permission Statement. Applicable to SBIR/STTR Only. See SBIR/STTR instructions. CHYes EH No PHS 398 (Rev. 09/04) Page 3 Form Page 2-continued Number the following pages consecutively throughout the application. Do not use suffixes such as 4a, 4b. Principal Investigator/Program Director (Last, First, Middle): Lyons, Karen M The name of the principal investigator/program director must be provided at the top of each printed page and each continuation page. RESEARCH GRANT TABLE OF CONTENTS Page Numbers Face Page 1 Description,

该建议研究了骨形态发生蛋白（BMP）在软骨发生中的作用。通过 对缺乏BMPR1A和/或BMPR1B的小鼠的分析，我们证明了通过这些受体的信号传导 对于软骨细胞增殖，分化和维持至关重要。这些研究表明 BMPR1A和BMPR1B在冷凝阶段具有重叠的功能，但没有揭示 他们在后来的阶段与之合作。而且，尽管ACTRI本身不足以支持软骨 创世纪以外的凝结阶段，BMPR1A和/或BMPR1B的程度可以促进 软骨发生尚不清楚。解决这个问题对于针对目标的未来策略很重要 激活特定的BMP受体，以引起组织工程应用中定义的响应或 在体内维修中。我们在AIM 1通过表征ACTRI/BMPRLA和 ACTRI/BMPRLB双重突变体，并通过BMP受体功能丧失的小鼠产生 可以在妊娠晚期诱导。 BMP可能通过 SMADS，但尚不清楚软骨发生是否需要所有3个BMP特异性SMAD或 SMAD是否具有不同的与重叠功能。我们在目标两个中解决了这个问题。研究 BMPR1A/1B化合物突变体发现了BMP信号丢失的证据伴随着增加 FGF途径的输出。这一发现表明，BMP和FGF途径在 软骨发生，并提高了BMP和FGF信号之间的相对平衡是一个 软骨细胞进展的主要决定因素通过生长板。我们测试并确定 FGF信号传导途径的下游组件在AIM三通过 构造BMP和FGF途径相对输出的小鼠的构建，并通过使用 肢体文化。 BMP受体突变体的分析表明，BMP途径对于 IHH的表达，IHH是软骨细胞增殖和分化生长板的关键调节剂。它有 在体外显示ERK1/2通过直接磷酸化拮抗BMP信号，从而使 灭活，BMP特异性SMAD。在目标四，我们测试该机制是否与IHH表达相关 在生长板中。了解BMP如何控制软骨发生的不同方面以及FGF 拮抗这些效果对于理解如何最佳利用BMP在组织中至关重要 工程应用，软骨维修和维护，并有可能治疗 由激活FGF突变引起的软骨发育不全。 加利福尼亚大学洛杉矶 大卫·格芬医学院 骨科外科 2614麦克唐纳研究实验室 675 Charles E Young Dr. South博士 洛杉矶，加利福尼亚州90095 PHS 398（Rev.09/04）第2页表格Page 2 首席调查员/计划主管（最后，第一，中间）： 关键人员。请参阅说明。根据需要使用延续页面 从首席研究员开始。按字母顺序列出所有其他关键人员，首先姓氏。 名称ERA CONSONS用户名组织 Karen Lyons 2 UCLA 肖恩·布鲁格加州大学洛杉矶分校 其他重要的贡献者 名称组织 Akiko Hata Tufts大学 大卫·奥尼茨华盛顿大学 里昂，卡伦 以下如下所示的信息。 项目的角色 pi 博士后研究员 项目的角色 顾问 顾问 人类胚胎干细胞E3否是是 如果提出的项目涉及人类胚胎干细胞，则从以下列表中列出了特定细胞LLNE的注册号以下： http：//stemcells.nih.qov/reqistrv/index.asp。根据需要进行用户关注页。 如果目前无法引用特定行，请包括一条说明将使用注册表的声明。 细胞系 披露许可声明。仅适用于SBIR/STTR。请参阅SBIR/STTR说明。 chyes eh no PHS 398（修订版09/04）第3页表格第2页。 在整个过程中连续编号以下页面 应用程序。请勿使用4A，4B等后缀。 首席研究员/计划主任（最后，第一，中间）：里昂，卡伦·M 必须在每个印刷页面的顶部和每个延续页面的顶部提供主要调查员/计划主管的名称。 研究赠款 目录 页码 面部第1页 描述，