The regulatory role of Bone morphogenetic protein 1 in suture patency and fusion

骨形态发生蛋白1在缝线通畅和融合中的调节作用

基本信息

批准号：
8733243
负责人：
Matthew P Harris
金额：
$ 35万
依托单位：
BOSTON CHILDREN'S HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-09-19 至 2014-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8733243
关键词：
Address Adult Affect Alleles Biochemical Biological Assay Bone Morphogenetic Proteins Chemicals Congenital abnormal Synostosis Craniosynostosis Data Defect Development Disease Dysmorphology Educational process of instructing Etiology Experimental Models Extracellular Matrix Extracellular Matrix Proteins Fishes Generations Genes Genetic Genetic Models Genetic Screening Goals Human Impaired cognition Integrins Joint structure of suture of skull Knowledge Lead Live Birth Maintenance Maps Mental Retardation Methods Microspheres Modeling Modification Molecular Genetics Mus Mutation Organism Participant Pathologic Pathology Pathway interactions Patients Pattern Peptide Hydrolases Phenotype Post-Translational Protein Processing Post-Translational Regulation Process Regulation Role Signal Transduction Skeletal Development Surgical sutures Testing Therapeutic Time Tissues Treatment Efficacy Vertebrates Welding Zebrafish base constriction cranium design implantation improved in vivo insight malformation mouse model mutant new therapeutic target novel null mutation procollagen C-endopeptidase public health relevance research study skeletal small molecule suture fusion treatment strategy

项目摘要

DESCRIPTION (provided by applicant): The correct patterning and timing of the fusion of cranial sutures is essential for normal skull development. Defects in suture formation lead to malformations and deformations, which cause constriction of the cranium and can impair cognition. Our knowledge of the mechanistic bases that distinguish normal from abnormal suture development is incomplete. The zebrafish is a powerful genetic and experimental model for exploring fundamental processes in vertebrate skeletal development. We have performed forward genetic screens in zebrafish to identify adult fish with defects in suture formation that are relevant to defects occurring in human patients. To efficiently map and clone the responsible genes, we developed new massively parallel sequencing and analytic strategies. This enabled us to identify bone morphogenetic protein 1a (bmp1a) and integrin ¿10 (itg¿10) as important participants during suture formation. Identifying genes involved in the post- translational modification of extracellular matrix (ECM) proteins and in the signaling that occurs between the ECM and the cell suggests changes in ECM and ECM-regulated cell signaling may comprise a shared downstream pathway for many genetic and environmental causes of abnormal suture development/closure. The goal of this proposal is to investigate the role of ECM post-translational modification and, specifically, the role of Bmp1 protease during normal and pathological suture formation. We will achieve these goals by completing 3 Aims. In Aim 1 we will identify new genetic regulators of suture formation by using mutants deficient in bmp1a to screen for dominant modifiers of Bmp1a function. Our preliminary data indicate this is a highly productive strategy. In Aim 2 we will characterize the effects of impaired Bmp1 activity on suture ECM proteins in zebrafish and determine whether altered modification of ECM proteins also occurs in synostoses from human patients. We have access to surgically excised affected and adjacent normal suture tissue from patients with known syndromic and unknown causes of synostosis. In Aim 3 we will capitalize on the zebrafish model's ability to be effectively utilizedin small molecule screens. We will specifically use bmp1a mutant fish to identify small molecules that alter Bmp1 activity in vivo. Molecules that affect the earliest phenotype observed in bmp1a mutant fish (altered finfolds) can then be tested for therapeutic efficacy in the late onset-phenotype (synostosis) using localized chemical- impregnated microbead implantation assays. The strength of the zebrafish model is that it enables the efficient identification of genes and pathways important for suture formation using forward genetic methods. Genes found to affect sutures in the zebrafish can then be studied in mammalian models of synostosis. Zebrafish are also useful for high-throughput in vivo screens for small molecule compounds that can inhibit or potentiate suture fusion. Compounds that alter deleterious phenotypes in zebrafish with Bmp1a mutations may have therapeutic value in patients with synostosis or with BMP1-associated phenotypes.

描述（由应用程序提供）：颅缝合融合的正确图案和时机对于正常的头骨发育至关重要。缝合形成的缺陷导致畸形和变形，这会导致颅骨收缩并可能损害认知。我们对将正常与异常缝合线发育区分开的机械碱基的了解是不完整的。斑马鱼是一个强大的遗传和实验模型，用于探索脊椎动物骨骼发育中的基本过程。我们已经在斑马鱼中进行了前遗传筛选，以鉴定成缝合形成缺陷的成年鱼，这些鱼与人类患者发生的缺陷有关。为了有效地映射和克隆负责任的基因，我们开发了新的大规模平行测序和分析策略。这使我们能够将骨形态发生蛋白1a（BMP1A）和整联蛋白€10（ITG？10）鉴定为成功形成过程中的重要参与者。识别涉及细胞外基质（ECM）蛋白的翻译后修饰的基因以及ECM和该细胞之间发生的信号传导表明，ECM和ECM调节的细胞信号的变化可能构成了许多遗传和环境原因的共享下游途径，这是许多遗传和环境因素成功开发/封闭/封闭的封闭/封闭。该提案的目的是研究ECM翻译后修饰的作用，特别是BMP1蛋白酶在正常和病理成功形成中的作用。我们将通过完成3个目标来实现这些目标。在AIM 1中，我们将通过使用BMP1A中的突变体确定成功形成的新遗传调节剂来筛选BMP1A功能的主要修饰符。我们的初步数据表明这是一种高产策略。在AIM 2中，我们将表征BMP1活性受损对斑马鱼中成功ECM蛋白的影响，并确定ECM蛋白的修饰改变是否也发生在人类患者的平稳性中。我们可以从已知综合症和未知原因的患者中获得手术影响受影响和邻近的正常成功组织。在AIM 3中，我们将利用斑马鱼模型有效利用小分子筛选的能力。我们将专门使用BMP1A突变鱼类来鉴定在体内改变BMP1活性的小分子。然后，可以使用局部化学浸渍的微粒植入分析测试影响BMP1A突变鱼类中最早表型的分子（更改鳍片）。斑马鱼模型的强度是，它可以有效地识别使用正向遗传学方法对缝合形成重要的基因和途径。然后，发现影响斑马鱼中缝合线的基因可以在哺乳动物模型的阴影模型中研究。斑马鱼对于可以抑制或潜在的缝合融合的小分子化合物的体内筛选也很有用。改变斑马鱼中有害表型具有BMP1A突变的化合物可能在冲突患者或与BMP1相关表型的患者中具有治疗价值。