Project 2. FKBP10 CHAPERONE COMPLEX AND TRAFFICKING AND ORGANELLE DYSFUNCTION IN

项目 2. FKBP10 伴侣复合体与贩运和细胞器功能障碍

• 批准号: 8380628
• 负责人: Brendan Lee
• 金额: $ 32.52万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8380628
• 关键词: Affect; Autophagocytosis; Autophagosome; Binding; Cartilage; Cells; Collagen Type I; Complex; Contracture; Data; Defect; Deformity; Diagnostic tests; Disease; Extracellular Matrix; Family; Fibroblasts; Fracture; Functional disorder; Genes; Genetic; Genetic Counseling; Goals; Golgi Apparatus; Growth; Homeostasis; Hsp47 protein; Human; Inherited; Instruction; Joints; Knockout Mice; Knowledge; Ligaments; Link; Maintenance; Maps; Mesenchymal; Molecular; Molecular Chaperones; Molecular Target; Mus; Mutation; Organelles; Osteogenesis; Osteogenesis Imperfecta; Outcome; Pain; Pathogenesis; Pathway interactions; Phenotype; Principal Investigator; Process; Proteins; Research; Role; Skeleton; Stress; Tendon structure; Testing; Tissues; Type I Procollagen; Work; base; bone; exome; improved; in vivo; insight; loss of function mutation; mineralization; novel; programs; skeletal; skeletal disorder; trafficking

Understanding key regulators of normal skeletal function have been facilitated by the study of osteogenesis imperfecta (01) or brittle bone disease. Our genetic studies have shown that humans with loss of function mutations in FKBPIO (which encodes the FKBP65 protein) have progressive deforming 01, which may also be associated with progressive joint contractures. What remains unresolved, but necessary to understand, are the cellular consequences and molecular interactions by which FKBP65 regulates synthesis of type I procollagen and formation of a proper bone extracellular matrix (ECM) leading to normal mineralization. The Specific Aims are: 1. Determine the consequences of FKBP10 loss on cellular phenotype. Loss of FKBP65 has significant effects on cellular phenotype indicating increased ER stress. We will determine the molecular basis for these observations in human 01 cells. 2. Determine the in vivo role of FKBP65 in mesenchymally derived tissues and determine its interaction with other ER-localized proteins. FkbpIO conditional knockout mice have been generated to determine whether the abnormal human cellular phenotype is recapitulated in the mouse. Based on the human phenotype, we will determine tissue-specific roles for Fkbp65 in the FkbpIO null mice.. 3. Determine the molecular basis for other recessive forms of 01. Using autozygosity mapping we have identified three new loci for recessively inherited forms of 01. We hypothesize that, like most genes associated with 01, these genes will be involved in the processing of type I procollagen and that their identification will reveal additional components essential for bone formation. We will use exome sequencing of genes in the autozygous regions to define these new 01 associated genes. The work proposed will determine the role of FKBP10/FKBP65 in mesenchymal tissues and will define new components necessary for normal bone formation. Overall the data generated will determine the molecules necessary for the synthesis, mineralization and maintenance of a normal type I collagen extracellular matrix. RELEVANCE (See instructions): The results of this project will have an immediate impact by providing improved genetic counseling and diagnostic testing for families with brittle bone disease. Determining the mechanism of disease in these new forms of Ol develop will identify disease-based strategies and specific molecular targets for therapy.

成骨研究有助于了解正常骨骼功能的关键调节因子 不完美 (01) 或脆骨病。我们的基因研究表明，功能丧失的人类 FKBPIO（编码 FKBP65 蛋白）的突变具有进行性变形 01，这也可能 与进行性关节挛缩有关。尚未解决但有必要理解的事情， FKBP65 调节 I 型合成的细胞后果和分子相互作用 前胶原和适当的骨细胞外基质（ECM）的形成导致正常矿化。这 具体目标是： 1. 确定 FKBP10 缺失对细胞表型的影响。 FKBP65 的丢失具有显着的 对细胞表型的影响表明内质网应激增加。我们将确定这些的分子基础 在人类01细胞中的观察结果。 2.确定FKBP65在间充质衍生组织中的体内作用并确定其与 其他内质网定位蛋白。已生成 FkbpIO 条件敲除小鼠以确定是否 异常的人类细胞表型在小鼠中得到重现。根据人类表型，我们 将确定 Fkbp65 在 FkbpIO 缺失小鼠中的组织特异性作用。 3. 确定 01 其他隐性形式的分子基础。使用自合性作图，我们得到 确定了 01 隐性遗传形式的三个新位点。我们假设，像大多数基因一样 与 01 相关，这些基因将参与 I 型前胶原的加工，并且它们的 鉴定将揭示骨形成所必需的其他成分。我们将使用外显子组测序 自合区域中的基因来定义这些新的 01 相关基因。 拟议的工作将确定 FKBP10/FKBP65 在间充质组织中的作用，并将定义新的 正常骨形成所必需的成分。总的来说，生成的数据将确定分子 正常 I 型胶原细胞外基质的合成、矿化和维持所必需的。 相关性（参见说明）： 该项目的结果将通过提供改进的遗传咨询和 对患有脆骨病的家庭进行诊断测试。确定这些新疾病的发病机制 Ol 的开发形式将确定基于疾病的策略和特定的治疗分子靶点。