Cell-cell adhesion in regulation of mammalian palatogenesis

哺乳动物腭发育调节中的细胞间粘附

• 批准号: 9328402
• 负责人: Kendall Lough
• 金额: $ 3.27万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-03-01 至 2019-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9328402
• 关键词: Actin-Binding Protein; Adaptor Signaling Protein; Adherens Junction; Adhesions; Affect; Americas; Apical; Apoptosis; Award; Binding; Binding Sites; Biological Assay; Biological Models; Biomedical Research; Candidate Disease Gene; Cell Adhesion; Cell Adhesion Molecules; Cell Line; Cell Proliferation; Cell-Cell Adhesion; Cells; Cellular biology; Cessation of life; Cleft Lip; Cleft Palate; Communication; Communities; Complex; Computational Biology; Congenital Abnormality; Data; Defect; Developing Countries; Developmental Biology; Disease; Dominant-Negative Mutation; E-Cadherin; Embryo; Ensure; Environment; Epidermis; Epithelial; Epithelium; Extracellular Domain; FURIN gene; Family; Fellowship; Gastrointestinal tract structure; Gene Expression; Gene Silencing; Genes; Genetic; Genetic Models; Genetic Techniques; Genetic study; Genomics; Goals; Human; In Vitro; Individual National Research Service Award; Knock-out; Link; Live Birth; Maintenance; Mammalian Genetics; Maps; Mediating; Mentors; Microscopy; Mitotic spindle; Modeling; Modernization; Morbidity - disease rate; Morphogenesis; Mus; Mutation; Nature; Nonsense Mutation; Online Mendelian Inheritance In Man; Oral; Oral cavity; Oral mucous membrane structure; PVRL1; Palate; Pathology; Pathway interactions; Phenotype; Physiology; Play; Population; Prevalence; Proteins; Recruitment Activity; Regulation; Research; Research Personnel; Role; South American; Study models; Syndrome; System; Techniques; Technology; Tight Junctions; TimeLine; Tissues; Ultrasonography; Vinculin; Writing; afadin; alpha catenin; career; cell cortex; craniofacial; experience; experimental study; gene discovery; genetic linkage analysis; genome wide association study; in vivo; interest; keratinocyte; knock-down; mouse model; mutant; nectin; novel; oral biology; oral cavity epithelium; orofacial cleft; palatal fusion; palatogenesis; pre-doctoral; prevent; skills; statistics; success; tool

ABSTRACT This application is for a Ruth L. Kirschstein National Research Service Award (NRSA) Individual Predoctoral Fellowship (F31) for Kendall J. Lough to study the role that the nectin-afadin cell adhesion pathway plays in regulating mammalian palatogenesis. Mr. Lough has demonstrated that loss of afadin results in highly penetrant cleft palate in mouse models. This award will allow Mr. Lough to further develop an expertise in cell and developmental biology while expanding his understanding in craniofacial morphogenesis and oral biology. This proposal will expand the understanding of mammalian palatogenesis by describing a novel cleft palate model while developing an additional model of an uncharacterized human cleft lip and cleft palate (CL/P) syndrome. Mr. Lough will take full advantage of the opportunities presented with this award to further develop skills essential to his career including 1) scientific communication through writing and oral presentation, 2) statistics and computational biology, 3) physiology in cell and developmental biology, 4) additional techniques relevant to biomedical research, and 5) significant mentoring experience. Mr. Lough has assembled a strong environment and research mentoring team to ensure the success of this proposal and his career goals. This award will prepare Mr. Lough to continue in his pursuit of a career as an independent academic researcher studying the genetics and cell biology of the oral mucosa, gastrointestinal tract and associated pathologies. Human CL/P is the most common birth defect in America and results in thousands of deaths globally each year, particularly in developing nations. While modern sequencing studies have identified numerous candidate loci associated with human CL/P, very few of these have been functionally characterized. One such disorder is CLPED1, which is associated with homozygous nonsense mutations in the extracellular domain of the nectin-1 encoding gene, PVRL1. Nectin-1 is part of the larger nectin family of transmembrane cell adhesion molecules, which bind to the cytoplasmic adapter protein afadin. Current mouse models lacking these genes are either embryonic lethal or fail to develop CL/P. Utilizing a novel genetic technique to inactivate gene expression via lentiviral delivery to early mouse embryos, Mr. Lough has demonstrated that loss of afadin results in CL/P, providing the first functional data linking this pathway to mammalian palatogenesis. This technique allows Mr. Lough to generate additional genetic mouse models at an unprecedented rate – 6 weeks as opposed to 6 months for standard techniques. This proposal outlines experiments to further characterize the mechanism of CL/P presentation in afadin null embryos through a combination of in vivo and in vitro techniques to evaluate changes in cell-cell adhesion. This application also proposes to generate an elegant genetic model expressing the human CL/P – associated Pvrl1W185X mutation in mice. This proposal will provide the scientific community with novel genetic tools and models for studying orofacial clefting candidates in an epithelial-specific manner.

摘要 这份申请是露丝·L Kirschstein国家研究服务奖（NRSA）个人 Kendall J. Lough博士前奖学金（F31），研究nectin-afadin细胞粘附途径的作用 在调节哺乳动物腭发育中起作用。洛夫先生已经证明，失去阿法丁的结果是高度 小鼠模型中的渗透性腭裂。该奖项将使Lough先生能够进一步发展细胞方面的专业知识。 和发育生物学，同时扩大他在颅面形态发生和口腔生物学的理解。 这一提议将通过描述一种新的腭裂来扩展对哺乳动物腭裂发生的理解 模型，同时开发了一个额外的模型，一个未知的人类唇腭裂（CL/P） 综合征Lough先生将充分利用这一奖项所提供的机会， 对他的职业生涯至关重要的技能，包括1）通过写作和口头陈述进行科学交流，2） 统计学和计算生物学，3）细胞生理学和发育生物学，4）其他技术 相关的生物医学研究，和5）重要的指导经验。拉夫先生召集了一个强大的 环境和研究指导团队，以确保这一建议的成功和他的职业目标。这 该奖项将准备洛夫先生继续在他的追求作为一个独立的学术研究人员的职业生涯 研究口腔粘膜、胃肠道和相关病理的遗传学和细胞生物学。 人类CL/P是美国最常见的出生缺陷，导致全球数千人死亡 尤其是在发展中国家。虽然现代测序研究已经确定了许多 与人类CL/P相关的候选基因座，其中很少有功能特征。一个这样 疾病是CLPED 1，其与细胞外结构域中的纯合无义突变相关， nectin-1编码基因PVRL 1。Nectin-1是跨膜细胞粘附的较大Nectin家族的一部分 分子，其与细胞质衔接蛋白afadin结合。目前缺乏这些基因的小鼠模型 利用一种新的遗传技术， 通过慢病毒传递到早期小鼠胚胎表达，Lough先生已经证明， 结果CL/P，提供了第一个功能数据连接这一途径哺乳动物腭发育。这 这项技术使Lough先生能够以前所未有的速度产生更多的遗传小鼠模型-- 6周 与标准技术的6个月相比。该提案概述了进一步表征 通过体内和体外结合，在afadin无效胚胎中CL/P呈递的机制 技术来评估细胞-细胞粘附的变化。该应用程序还建议生成一个优雅的 在小鼠中表达人CL/P相关Pvr 11 W185 X突变的遗传模型。该提案将提供 科学界用新的遗传工具和模型研究口面裂候选人， 上皮特异性方式。