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 此申请适用于 Ruth 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 在调节哺乳动物的腭发育中发挥作用。 Lough 先生已经证明，afadin 的丢失会导致高度 小鼠模型中的穿透性腭裂。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，同时扩大他对颅面形态发生和口腔生物学的理解。 该提案将通过描述一种新颖的腭裂来扩大对哺乳动物腭裂的理解 模型，同时开发未表征的人类唇裂和腭裂 (CL/P) 的附加模型 综合症。 Lough先生将充分利用该奖项提供的机会进一步发展 对他的职业生涯至关重要的技能，包括 1) 通过写作和口头演示进行科学交流，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 与生物医学研究相关，以及 5) 丰富的指导经验。洛夫先生集结了强大的 环境和研究指导团队确保了这个提案的成功和他的职业目标。这 该奖项将为 Lough 先生继续追求独立学术研究员的职业生涯做好准备 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 是美国最常见的出生缺陷，导致全球数千人死亡 每年，特别是在发展中国家。虽然现代测序研究已经发现了许多 与人类 CL/P 相关的候选基因座，其中很少有功能特征。这样的一位 疾病是 CLPED1，它与细胞外域的纯合无义突变相关。 nectin-1 编码基因 PVRL1。 Nectin-1 是跨膜细胞粘附的 nectin 家族的一部分 分子，与细胞质接头蛋白 afadin 结合。目前的小鼠模型缺乏这些基因 要么胚胎致死，要么无法发育 CL/P。利用新型遗传技术灭活基因 通过慢病毒传递至早期小鼠胚胎进行表达，Lough 先生证明了 afadin 的丢失 CL/P 的结果，提供了将该途径与哺乳动物腭发育联系起来的第一个功能数据。这 技术使 Lough 先生能够以前所未有的速度（6 周）生成额外的基因小鼠模型 而标准技术则需要 6 个月。该提案概述了进一步表征的实验 通过体内和体外结合在 afadin 无效胚胎中 CL/P 呈递机制 评估细胞间粘附变化的技术。该应用程序还建议生成一个优雅的 在小鼠中表达人类 CL/P 相关 Pvrl1W185X 突变的遗传模型。该提案将提供 科学界拥有新颖的遗传工具和模型来研究口颌面裂候选者 上皮特异性方式。