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.

抽象的 该申请是露丝·柯希斯坦国家研究服务奖（NRSA）个人 Kendall J. Lough研究nectin-Afadin细胞粘附途径的作用 在调节哺乳动物的古质发生中发挥作用。拉夫先生已经证明，afadin的损失高度导致 小鼠模型中的渗透裂纹口感。该奖项将使Lough先生能够进一步发展细胞专业知识 和发育生物学，同时扩大了他对颅面形态发生和口服生物学的理解。 该提议将通过描述一种新颖的口感来扩展对哺乳动物古质发生的理解 在开发未表征的人裂唇和left裂（Cl/p）的附加模型的同时 综合征。 Lough先生将充分利用该奖项的机会，以进一步发展 对他的职业至关重要的技能，包括1）通过写作和口头演讲的科学交流，2） 统计和计算生物学，3）细胞和发育生物学的生理学，4）其他技术 与生物医学研究有关，以及5）重要的心理经验。拉夫先生聚集了一个强大的 环境和研究指导团队，以确保该提案的成功及其职业目标。这 奖项将为湖泊准备继续从事独立学术研究员的职业 研究口腔粘膜，胃肠道和相关病理学的遗传学和细胞生物学。 人类CL/P是美国最常见的先天缺陷，在全球范围内导致数千人死亡 每年，特别是在发展中国家。现代测序研究已经确定了许多 与人Cl/p相关的候选区域，其中很少有功能表征。一个这样的 疾病是clped1，它与纯合废话突变有关 Nectin-1编码基因PVRL1。 Nectin-1是跨膜细胞粘附的较大的花蜜家族的一部分 分子，与细胞质衔接蛋白afadin结合。当前缺乏这些基因的鼠标模型 要么是胚胎致死的，要么无法发育Cl/p。利用一种新型遗传技术灭活基因 通过慢病毒递送到早期小鼠胚胎的表达，Lough先生证明了Afadin的丧失 导致Cl/p，提供了将此途径与哺乳动物palategenation联系起来的第一个功能数据。这 技术允许Lough先生以前所未有的速度生成其他遗传小鼠模型 - 6周 与标准技术的6个月相反。该建议概述了实验，以进一步表征 通过体内和体外的组合，Afadin Null胚胎中Cl/P呈现的机理 评估细胞细胞粘附变化的技术。该应用程序还建议生成优雅 表达人类CL/P - 相关的PVRL1W185X突变的遗传模型。该建议将提供 具有新颖的遗传工具和模型的科学界，用于研究候选者 上皮特异性方式。