Genetic Analysis of Bone Morphogenic Proteins

骨形态发生蛋白的遗传分析

• 批准号: 6797878
• 负责人: DAVID M KINGSLEY
• 金额: $ 33.84万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1993
• 资助国家: 美国
• 起止时间: 1993-08-01 至 2007-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6797878
• 关键词: SDS polyacrylamide gel electrophoresis; alleles; biological signal transduction; bone development; bone morphogenetic proteins; bone regeneration; cartilage development; developmental genetics; expression cloning; gene expression; gene mutation; gene targeting; genetically modified animals; joints; laboratory mouse; molecular cloning; musculoskeletal regeneration; nucleic acid sequence; phenotype; polymerase chain reaction; protein structure function; transcription factor

DESCRIPTION (provided by applicant): Bone morphogenetic proteins (BMPs) are key signalling molecules that control many different steps in development. A large number of studies suggest that BMPs are the endogenous signals used by vertebrates to induce formation of cartilage and bone during embryonic development, and to stimulate repair of bone fractures in adults. More recent studies suggest that some BMP family members also play an important role in joint formation, and that BMP signalling may be required to maintain articular cartilage after birth. The remarkable ability of these proteins to induce the formation of new skeletal tissues when implanted at ectopic sites raises the possibility that they may form the basis of important new clinical treatments for fracture repair, spinal fusion, increased bone strength, and maintenance of articular cartilage. Despite the importance of BMPs in skeletal biology, very little is known about the molecular mechanisms that control where and when they are normally expressed. Increased understanding of this area may suggest new ways of manipulating BMP expression at particular sites and stages of development, or to inhibit BMP expression in diseases of ectopic bone formation. We are carrying out a detailed genetic, genomic, and functional study of the regulatory sequences that control expression of 3 different BMP genes during normal development. These studies have already shown that the regulatory sequences that control BMP expression are distributed over enormous distances around the coding sequences. The overall patterns of BMP expression are built up from a number of modular elements, each responsible for controlling expression in a small anatomical subset of the overall skeleton. We will use a combination of comparative sequencing, clone scanning, and functional tests in transgenic mice to identify minimal control elements responsible for BMP expression in the key proliferative layer that surrounds growing skeletal elements, and in interzones that mark the sites of joint formation. Transcription factors that act through these elements will be identified by a combination of expression screening and biochemical interaction with wild-type and mutant control elements. The isolated control elements will also be used to test the functional role of BMP signalling in controlling the size, shape, and curvature of ribs. Finally, joint specific control elements will be used to develop a general system for studying the role of other genes in joints.

描述（由申请人提供）：骨形态发生蛋白（BMP）是控制许多不同发展步骤的关键信号分子。大量研究表明，BMP是脊椎动物在胚胎发育过程中诱导软骨和骨骼形成的内源信号，并刺激成人骨折的修复。最近的研究表明，一些BMP家族成员在联合形成中也起着重要作用，并且可能需要BMP信号传导才能在出生后保持关节软骨。当在异位部位植入时，这些蛋白质的显着能力诱导新骨骼组织的形成，这使它们可能形成了重要的新临床治疗方法的基础，以进行断裂修复，脊柱融合，增加骨骼强度和维持关节软骨的维持。尽管BMP在骨骼生物学中的重要性，但对控制通常表达的何时何地的分子机制知之甚少。对该领域的了解增加可能表明在特定部位和发育阶段操纵BMP表达的新方法，或抑制异位骨形成疾病中BMP表达。我们正在对正常发育过程中控制3种不同BMP基因的调控序列进行详细的遗传，基因组和功能研究。这些研究已经表明，控制BMP表达的调节序列分布在编码序列周围的巨大距离上。 BMP表达的总体模式是由许多模块化元素构建的，每个元素都负责控制整体骨骼的小解剖子集中的表达。我们将在转基因小鼠中使用比较测序，克隆扫描和功能性测试的组合，以识别负责BMP表达的最小对照元件，这些元件围绕着生长的骨骼元件的关键增殖层以及标记关节构成部位的骨骼中的重增强层。通过这些元素起作用的转录因子将通过表达筛选和生化相互作用与野生型和突变控制元件的结合来鉴定。孤立的控制元件还将用于测试BMP信号在控制肋骨的大小，形状和曲率中的功能作用。最后，将使用特定特异性控制元件来开发一种通用系统来研究其他基因在关节中的作用。