The Role of GSK-3beta in Palate Development and Fusion

GSK-3beta 在味觉发育和融合中的作用

• 批准号: 7674315
• 负责人: MICHAEL T LONGAKER
• 金额: $ 8.85万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-08-12 至 2009-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7674315
• 关键词: Address; Affect; Alleles; Animal Model; Apoptosis; Biological Markers; Biology; Case Study; Cell Death; Child; Cleaved cell; Cleft Palate; Clinical; Closure; Complex; Congenital Abnormality; Data; Defect; Dental Occlusion; Dentition; Development; Developmental Process; Dose; Drug Kinetics; Drug-sensitive; Ear; Embryonic Development; Environmental Risk Factor; Etiology; Evaluation; Event; Face; Family; Future; Genes; Genetic; Growth; Hospital Costs; Hour; Human; Human Development; In Vitro; Incidence; Investigation; Knowledge; Learning; Liquid substance; Live Birth; Location; Maxilla; Methods; Molecular; Mus; Mutant Strains Mice; Numbers; Obstruction; Operative Surgical Procedures; Organ Culture Techniques; Palate; Pathogenesis; Pathway interactions; Patients; Pattern; Pharmaceutical Preparations; Pharmacodynamics; Phenotype; Phosphotransferases; Positioning Attribute; Process; Proteins; Protocols documentation; Public Health; Regulation; Role; Secondary Palate; Signal Pathway; Signal Transduction; Signal Transduction Pathway; Skeleton; Source; Speech; Staging; Structure; System; Techniques; Technology; Time; Treatment Protocols; base; cell growth; craniofacial; cranium; critical developmental period; drug sensitivity; epithelial to mesenchymal transition; experience; feeding; genetic analysis; glycogen synthase kinase 3 beta; in utero; in vivo; mutant; novel; orofacial; palatal shelves; palatogenesis; prevent; psychosocial; research study; response; socioeconomics; tool; transdifferentiation

DESCRIPTION (provided by applicant): Craniofacial birth defects represent devastating psychosocial complications as well as a significant socioeconomic burden. Among them, cleft palate posed an annual "national bill" exceeding $100 million in 2004 in direct hospital costs alone and is highly prevalent as the incidence is roughly 0.1% (one in 1000 children), making it the second most common birth defect. Previous studies indicate that the pathogenesis of cleft palate is multifactorial and likely has both genetic and environmental factors. Much of our knowledge of craniofacial clefting arises from case studies of patients and although limited, selected animal models. A number of genes have been identified to be associated with the cleft palate phenotype, but the etiology of the majority of cases remains elusive. Affected children require multiple operations to address not only palate closure, but also associated problems with speech, feeding, dentition, and other craniofacial growth deficiencies. The function of GSK-32 has previously been shown to be intimately related to and necessary for normal craniofacial development. The central hypothesis of this application is that signal transduction pathways dependent on GSK-32 control the development of the secondary palate. This proposal will use genetic analysis and novel protein regulation techniques to study the roles of GSK-32 in palate formation. Two Specific Aims are proposed to explore the roles of GSK-32 in palatogenesis and fusion. The first aim will address the mechanistic requirements of GSK-32 signaling in palatogenesis while the second aim brings a new technique to control levels of GSK-32 protein to the study of palate development. In the first aim, the mechanism(s) underlying the cleft of the secondary palate in GSK-32 mutant mice will be explored. Palatal shelves in this mutant have previously been shown to develop seemingly appropriately during embryogenesis, but fail to fuse. This deficiency may be the result of defective palatal shelf growth, cell apoptosis and/or transdifferentiation, and the first part of this aim will look in vivo to investigate markers of these processes during palatogenesis in mutant compared to the wild-type palates. In the second part, in vitro organ culture will be used to determine if the mutant palateal shelves are capable of fusion when placed in apposition in organ culture. In the second aim, the study of palatogenesis using a drug-dependent GSK-32 allele during distinct stages will be undertaken. Because GSK-32 is required at different times in different regions, complete early loss of its activity may preclude study of its action at a downstream time and location for example, growth and fusion events in the palate after defective maxillogenesis. This system is novel and requires further investigation and adaptation to in vitro protocols. Taken together, these two aims will pinpoint the spatial and temporal requirements of GSK-32 signaling and greatly advance our knowledge of the mechanistic causes of human orofacial clefting. Furthermore, the new protein regulation techniques that we develop will serve as a template for future developmental studies. PUBLIC HEALTH RELEVANCE: Children and families affected by cleft palate must not only endure multiple, physiologically challenging surgeries to address palate closure, but also associated problems with speech, feeding, dentition, and other facial growth deficiencies. In addition, cleft palate brings with it devastating psychosocial implications for many children as well as significant socioeconomic burden exceeding $100 million in direct hospital costs alone. These facts are significant and while certain genes and environmental factors have been associated with the development of cleft palate, the cause in the majority of cases remains unknown and further study is extremely important.

描述（由申请人提供）：颅面的先天缺陷代表了毁灭性的社会心理并发症以及重大的社会经济负担。其中，Cleft Palete在2004年的直接医院费用上构成了一年一度的“国家账单”，高于1亿美元，并且非常普遍，因为发病率约为0.1％（其中1000名儿童中有1个），这使其成为第二大最常见的先天性缺陷。先前的研究表明，left裂的发病机理是多因素的，并且可能具有遗传和环境因素。我们对颅面裂裂的大部分知识来自对患者的病例研究，尽管有限，但选定的动物模型。已经确定了许多基因与left裂的表型相关，但是大多数病例的病因仍然难以捉摸。受影响的儿童需要多次操作，不仅可以解决口感关闭，还需要解决言语，喂养，牙齿和其他颅面生长不足的问题。 GSK-32的功能先前已被证明与正常的颅面发育密切相关，并且必不可少。该应用的中心假设是信号转导途径依赖于GSK-32控制次级口感的发展。该提案将使用遗传分析和新型蛋白质调节技术来研究GSK-32在pa层形成中的作用。提出了两个具体的目的，以探索GSK-32在古质发生和融合中的作用。第一个目的将解决gsk-32信号传导在古质发生中的机械要求，而第二个目标则带来了一种新技术，以控制GSK-32蛋白的水平，以研究味蕾的发展。在第一个目标中，将探索GSK-32突变小鼠中次生口感裂口的基础机制。以前，该突变体中的pal架架子在胚胎发生过程中似乎适当地发育，但无法融合。这种缺乏可能是pal质架架的生长，细胞凋亡和/或转分化的结果，与野生型味相比，该目标的第一部分将在体内观察到突变体的pala核发生过程中这些过程的标记。在第二部分中，将使用体外器官培养物来确定突变的palateal架子是否能够在器官培养中置于体内时能够融合。在第二个目标中，将在不同阶段使用药物依赖性GSK-32等位基因进行古质发生。由于在不同区域的不同时间需要GSK-32，因此其活性的完全早期丧失可能排除在下游时间和位置对其作用的研究，例如，上颌骨发生后味觉的生长和融合事件。该系统是新颖的，需要进一步研究和适应体外方案。综上所述，这两个目的将指出GSK-32信号传导的空间和时间要求，并大大提高了我们对人类口面裂的机械原因的了解。此外，我们开发的新蛋白质调节技术将作为未来发展研究的模板。 公共卫生相关性：受left口感影响的儿童和家庭不仅必须忍受多次，生理上具有挑战性的手术，以解决闭塞的问题，而且还必须与言语，喂养，牙齿和其他面部生长不足有关。此外，Cleft Palete为许多儿童带来了毁灭性的社会心理影响，仅需直接医院费用就超过了1亿美元的社会经济负担。这些事实是重要的，尽管某些基因和环境因素与left裂的发展有关，但大多数病例的原因仍然未知，进一步的研究非常重要。