Eda/Edar Regulation of Embryonic SMG Development

描述(申请人提供)：少汗性(无汗性)外胚层发育不良(HED)是约150种已描述的外胚层发育不良中最常见的，是一种以毛发、牙齿、汗腺和唾液腺异常发育为特征的疾病。EDA和EDAR(EDA受体)基因的突变分别导致X连锁和常染色体Hed；类似于Hed的异常表型在猫(eda-ta)和无绒(edar-dl)突变小鼠中被发现。鉴于在HED患者以及类似的猫(Ta)和Downless小鼠中观察到异常的颌下腺(SMG)，我们的免疫组化定位：EDA和EDAR蛋白在胚胎SMG上皮细胞上的定位，以及我们的发现在体外增强或干扰EDA/EDAR信号调节颌下腺异常分支的形态发生，我们假设ECLA/EDAR信号转导在SMG的发育中至关重要。为了阐明EDA/EDAR信号在胚胎SMG发育中的作用，我们建议：(A)确定胚胎Ta突变SMG中取消的EDA/EDAR信号的进行性、阶段依赖性的表型结果，并将其与野生型(WT)和基因恢复的猫-EDA-A1转基因小鼠SMG中的结果进行比较。SMG的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，描述胚胎Ta SMGs中缺失的EDA/EDAR/NFkappaB信号的功能基因组学，并与WT和猫转基因小鼠SMGs进行比较。(C)通过比较胚胎Ta、WT和猫-EDA-A1 SMG中激活蛋白表达的变化，确定EDA/EDAR/NFkappaB信号下游直接或侧面的功能通路。最后，我们将描绘激活蛋白在体内的时空分布：受中断的EDA/EDAR/NFkappaB信号的调节。我们提出的研究数据将为未来SMG功能丧失的研究提供框架，并将为设计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。

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.