NOVEL SCREEN FOR MINERALIZED CRANIOFACIAL AND TOOTH MUTANTS IN ZEBRAFISH

斑马鱼矿化颅面和牙齿突变体的新型筛查

• 批准号: 7191891
• 负责人: PAMELA C YELICK
• 金额: $ 39.65万
• 依托单位: TUFTS UNIVERSITY BOSTON
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-02-01 至 2010-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7191891
• 关键词: Animal Model; Award; Binding; Biological Assay; Biological Response Modifier Therapy; Cataloging; Catalogs; Chemicals; Cleft Lip; Complex; Complication; Craniosynostosis; DNA; Data; Defect; Development; Ethylnitrosourea; Exhibits; Family; Gene Mutation; Genes; Genetic; Genomics; Goals; Grant; Growth and Development function; Human; Human Development; In Vitro; Joint structure of suture of skull; Knowledge; Laboratories; Left; Life; Mammals; Maps; Methods; Modeling; Molecular; Molecular Genetics; Molecular Models; Molecular Target; Mus; Mutagenesis; Natural regeneration; Nature; Operative Surgical Procedures; Palate; Pathology; Pathway interactions; Penetrance; Personal Satisfaction; Proteins; Range; Research; Research Design; Research Personnel; Sample Size; Signal Transduction; Skeletal Development; Skeletal system; Statistically Significant; Supernumerary Tooth; Testing; Time; Tissue Sample; Tissues; Tooth Loss; Tooth structure; Urination; Work; Zebrafish; base; cartilage development; clinically relevant; craniofacial; craniofacial repair; design; experience; gene conservation; gene function; genetic analysis; human tissue; in vivo; innovation; mutant; novel; programs; receptor; repaired; suture fusion

DESCRIPTION (provided by applicant): The broad objective of this study is to identify molecular targets regulating zebrafish craniofacial skeletogenesis and primary and replacement tooth formation (RTF), that can be manipulated to repair human craniofacial skeletal and tooth defects in humans. Although significant research efforts have resulted in the identificaton of certain gene mutations responsible for human craniofacial skeletal defects, functional characterizations have been hampered by limited availability of human tissues and suitable animal models. Similarly, clinically relevant biological therapies to regenerate teeth do not currently exist, and research on tooth regeneration strategies is hampered by similar limitations. Targeted, molecular based therapies to effectively and permanently correct mineralized craniofacial and tooth defects would be a significant advancement over current surgical repair methods. The fact that zebrafish craniofacial skeletal development closely resembles that of mammals, combined with the fact that zebrafish continuously regenerate teeth throughout their lives, provides a unique opportunity to define and funcitionally characterize molecular signaling cascades directing mineralized craniofacial skeletal and replacement tooth development. The extensive conservation of gene identity, genomic organization and gene function between zebrafish and humans allows for studies in zebrafish to be directly related to human development. Our long term goal is to identify genetic pathways that can be manipulated to repair craniofacial skeletal defects and initiate RTF in humans. The objective of this study is to exploit the zebrafish to perform functional studies of molecular mechanisms regulating craniofacial mineralized skeletal and tooth formation. In this proposal we will first perform a forward genetic ENU mutagenesis screen, combined with a sensitive in vivo assay for mineralized tissue formation, to identify novel craniofacial skeletal and RTF mutants. Next, we will confirm the molecular identity, and perform functional characterization, of each identified mutant. We anticipate that the proposed studies will reveal molecular signaling cascades that may facilitate the implementation of clinically relevant gene based therapies to correct mineralized craniofacial skeletal and tooth defects in humans.

描述（由申请人提供）：本研究的主要目标是确定调节斑马鱼颅面骨骼发生以及乳牙和替换牙形成（RTF）的分子靶标，这些分子靶标可用于修复人类颅面骨骼和牙齿缺陷。尽管大量的研究工作已经确定了导致人类颅面骨骼缺陷的某些基因突变，但由于人体组织和合适的动物模型的可用性有限，功能表征受到了阻碍。同样，目前还不存在临床相关的牙齿再生生物疗法，并且牙齿再生策略的研究也受到类似限制的阻碍。有效且永久地纠正矿化颅面和牙齿缺陷的靶向分子疗法将是对当前手术修复方法的重大进步。斑马鱼颅面骨骼发育与哺乳动物非常相似，再加上斑马鱼在其一生中不断再生牙齿，这一事实为定义和功能表征指导矿化颅面骨骼和替代牙齿发育的分子信号级联提供了独特的机会。斑马鱼和人类之间基因特性、基因组组织和基因功能的广泛保守性使得斑马鱼研究能够与人类发育直接相关。我们的长期目标是确定可操纵修复颅面骨骼缺陷并启动人类 RTF 的遗传途径。本研究的目的是利用斑马鱼对调节颅面矿化骨骼和牙齿形成的分子机制进行功能研究。在本提案中，我们将首先进行正向遗传 ENU 诱变筛选，结合矿化组织形成的敏感体内测定，以鉴定新的颅面骨骼和 RTF 突变体。接下来，我们将确认每个已识别突变体的分子身份并进行功能表征。我们预计拟议的研究将揭示分子信号级联，这可能有助于临床相关基因疗法的实施，以纠正人类矿化颅面骨骼和牙齿缺陷。