Eda/Edar Regulation of Embryonic SMG Development

胚胎 SMG 发育的 Eda/Edar 调节

• 批准号: 6678568
• 负责人: TINA JASKOLL
• 金额: $ 39.29万
• 依托单位: UNIVERSITY OF SOUTHERN CALIFORNIA
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-07-01 至 2008-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6678568
• 关键词: RNase protection assay; acinar cell; apoptosis; biological signal transduction; cell proliferation; developmental genetics; ectoderm; functional /structural genomics; gene expression; genetic disorder; genetic regulation; genetically modified animals; laboratory mouse; mammalian embryology; membrane proteins; mucins; nuclear factor kappa beta; phenotype; proteomics; submandibular gland

DESCRIPTION (provided by applicant): Hypohidrotic (anhidrotic) ectodermal dysplasia (HED), the most common of the approximately 150 described ectodermal dysplasias, is a disorder characterized by abnormal development of hair, teeth, sweat glands and salivary glands. Mutations in the: EDA and EDAR (EDA receptor) genes are responsible for X-linked and autosomal HED, respectively; abnormal phenotypes similar to HED are seen in Tabby (Eda-Ta) and downless (Edar-dl) mutant mice. Given the observation of abnormal submandibular salivary glands (SMG) in patients with HED and, similarly, in Tabby (Ta) and downless mice, our immunolocalization:of Eda and Edar proteins on embryonic SMG epithelia, and that our finding enhanced or interrupted Eda/Edar signaling in vitro modulates SMG branching morphogenesis, we postulate that Ecla/Edar signal transduction is essential for SMG development. To delineate the role of Eda/Edar signaling during embryonic SMG development, WE PROPOSE to: (a) Determine the progressive, stage-dependent phenotypic outcome of abrogated Eda/Edar signaling in embryonic Ta mutant SMGs and compare it to that seen in wildtype (WT) and gene-restored Tabby-EDA-A1 transgenic mouse SMGs. SMG phenotypes will be defined by microanatomy, cell proliferation and apoptosis indices, level of NFkappaB activation, and mucin protein expression. (b) Delineate the functional genomics of abrogated Eda/Edar/NFkappaB signaling in embryonic Ta SMGs, as compared to WT and Tabby-EDA-A1 transgenic mouse SMGs, using transcriptomic and proteomic methodologies. (c) Identify the functional pathways directly or collaterally downstream of the Eda/Edar/NFkappaB signal by comparing changes in activated protein expression in embryonic Ta, WT, and Tabby-EDA-A1 SMGs. Finally, we will delineate the in vivo spatiotemporal distribution of activated proteins: modulated with interrupted Eda/Edar/NFkappaB signaling. The data generated by our proposed studies will provide the framework for future studies of SMG loss of function and will provide a rational basis for designing drugs that may be used to restore acinar form and function in damaged and aging SMGs.

描述（由申请人提供）：少汗性（无汗性）外胚层发育不良（HED）是大约150种外胚层发育不良中最常见的一种，是一种以头发、牙齿、汗腺和唾液腺发育异常为特征的疾病。EDA和EDAR （EDA受体）基因的突变分别导致x连锁HED和常染色体HED；在斑纹（Eda-Ta）和无down型（eda -dl）突变小鼠中可以看到与HED相似的异常表型。考虑到HED患者和Tabby （Ta）和down - less小鼠的下颌下唾液腺（SMG）异常，我们在胚胎SMG上皮上的Eda和Edar蛋白的免疫定位，以及我们在体外发现Eda/Edar信号增强或中断可调节SMG分支形态发生，我们假设Ecla/Edar信号转导对SMG发育至关重要。为了描述Eda/Edar信号在胚胎SMG发育过程中的作用，我们建议：(a)确定胚胎Ta突变SMG中Eda/Edar信号缺失的进展性、阶段依赖性表型结果，并将其与野生型（WT）和基因恢复tab比- Eda - a1转基因小鼠SMG进行比较。SMG表型将通过显微解剖、细胞增殖和凋亡指数、NFkappaB激活水平和粘蛋白表达来确定。(b)利用转录组学和蛋白质组学方法，与WT和Tabby-EDA-A1转基因小鼠SMGs相比，描述胚胎Ta SMGs中取消的Eda/Edar/NFkappaB信号的功能基因组学。(c)通过比较胚胎Ta、WT和Tabby-EDA-A1 SMGs中活化蛋白表达的变化，确定Eda/Edar/NFkappaB信号直接或间接下游的功能通路。最后，我们将描述激活蛋白的体内时空分布：被中断的Eda/Edar/NFkappaB信号调节。我们提出的研究产生的数据将为未来SMG功能丧失的研究提供框架，并将为设计可用于恢复受损和老化SMG的腺泡形态和功能的药物提供合理的基础。