CHD1, Chromatin Dynamics, and Salivary Gland Differentiation

CHD1、染色质动力学和唾液腺分化

• 批准号: 7867975
• 负责人: Joseph Adam Hall
• 金额: $ 3.52万
• 依托单位: MARSHALL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-06-01 至 2011-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7867975
• 关键词: Address; Affect; Aging; Alternative Splicing; Autoimmune Diseases; Autoimmune Process; Biological Assay; Biological Models; Cell physiology; Chromatin; DNA-Protein Interaction; Data; Deglutition; Dental caries; Development; Disease; Environmental Risk Factor; Epidemiology; Epigenetic Process; Exhibits; Frequencies; Gene Expression; Gene Expression Regulation; Genes; Gland; Glycoproteins; Goals; Immunoblotting; Immunoprecipitation; Individual; Investigation; Knowledge; Laboratories; Lacrimal gland structure; Light; Local Anti-Infective Agents; Mediating; Messenger RNA; Methods; Modeling; Molecular; Mucins; Mucous body substance; Mus; NFS/N Mouse; Neonatal; Organ; Phenotype; Phonation; Polymerase Chain Reaction; Production; Property; Proteins; Proteolysis; RNA Splicing; Regulator Genes; Role; Saliva; Salivary; Salivary Glands; Salts; Sjogren' s Syndrome; Sublingual Gland; Techniques; Transcript; Translations; Ulcer; Variant; Water; Xerostomia; chromatin remodeling; design; in vivo; new therapeutic target; novel; promoter; protein protein interaction; research study; saliva secretion; salivary mucins

DESCRIPTION (provided by applicant): Salivary gland differentiation and functionality relies on the production and secretion of saliva. Mucous, produced mainly by the sublingual glands (SLGs), confers viscous and antiseptic properties on saliva. Mucin glycoproteins are the major constituent of mucous, essential for the formation of mucosal barriers. Hyposecretion of mucins by the SLGs results in xerostomia, characterized by difficulty in phonation and deglutition, mucosal ulcerations, and an increase in the frequency of dental caries. One of the major contributors to xerostomia is an autoimmune disorder known as Sjogren's Syndrome which affects multiple body organs, most notably the salivary and lacrimal glands. NFS-sId mice, a model for Sjogren's Syndrome, display delayed SLG differentiation noted by neonatal mucin-deficiencies, specifically the major mucin product of mouse SLGs, Muc19. Interestingly, our laboratory has confirmed a function for the chromatin- remodeling protein CHD1 in regulating Muc19 expression. This project is designed to further investigate the molecular mechanisms that govern mucin production by the SLGs through the scope of CHD1 and its interactors, relating these findings to the development of novel therapeutic targets for re-activation of mucin production. To accomplish this, the following specific aims will be addressed: Aim 1: Compare levels of CHD1 and Mud9 expression, CHD1 protein levels, and the physical presence of CHD1 on the Muc19 promoter and transcribed regions, in NFS-sId and NFS-N (control) mice. Aim 2: Investigate the possibilities that the method of Muc19 gene regulation by CHD1 is occurring at the transcriptional elongation and/or alternative-splicing level. NFS-sId and control NFS-N mice will be utilized for all experiments. Gene expression and immunoblotting assays, combined with in vivo immunoprecipitation experiments, will shed light on the influence of quantity, localization, and partnership of CHD1 and its interactors on the expression, and subsequent production, of Muc19, while additional in vivo DNA-protein interaction studies, combined with an investigation of Muc19 transcript variants will allow inferences on the specific mechanism of Muc19 gene regulation in SLGs. The gene-regulatory mechanisms that control mucin production in the SLG remain poorly understood. Knowledge gained through the completion of this project will enhance the understanding of the epidemiology related to the hyposecretion of mucins, such as that which occurs in those afflicted with Sjogren's Syndrome. Also, novel therapeutic targets aimed at re-activation of SLG functionality could be identified.

描述（由申请人提供）：唾液腺分化和功能依赖于唾液的生产和分泌。粘液主要由舌下腺（SLG）产生，赋予唾液上的粘性和防腐特性。粘蛋白糖蛋白是粘液的主要成分，对于形成粘膜屏障必不可少。 SLGS对粘蛋白的分泌会导致静态症，其特征是发声和脱水，粘膜溃疡和牙齿龋齿频率的增加。造口症的主要因素之一是一种自身免疫性疾病，称为Sjogren综合征，它影响了多个身体器官，最著名的是唾液和泪腺。 NFS-SID小鼠是Sjogren综合征的模型，显示了新生儿粘蛋白缺陷指出的延迟SLG分化，特别是小鼠SLG的主要粘蛋白产物MUC19。有趣的是，我们的实验室已经证实了染色质重塑蛋白CHD1在调节MUC19表达中的功能。该项目旨在进一步研究SLG通过CHD1及其相互作用者粘蛋白产生的分子机制，将这些发现与新型治疗靶标的开发有关，以重新激活粘蛋白。为此，将解决以下特定目标：目标1：比较chd1和Mud9表达的水平，CHD1蛋白水平以及在MUC19启动子上的CHD1的物理存在，并在NFS-SID和NFS-N（对照）小鼠中比较了CHD1蛋白水平和转录区域。 AIM 2：研究CHD1调节MUC19基因调节方法的可能性是在转录伸长和/或替代性拼写水平上发生的。 NFS-SID和对照NFS-N小鼠将用于所有实验。基因表达和免疫印迹测定，结合体内免疫沉淀实验，将揭示CHD1的数量，定位和伙伴关系及其相互作用者对MUC19的表达，随后的产生的影响，而在VIVO DNA相互作用中的影响，允许MUC19的特定研究MUC19的转移，而MUC19的额外作用将与MUC19的特定性结合在一起。 SLG中的基因调节。控制SLG中粘蛋白产生的基因调节机制仍然鲜为人知。通过完成该项目获得的知识将增强对与粘蛋白降低相关的流行病学的理解，例如在患有Sjogren综合征的患者中发生的粘蛋白。同样，可以确定旨在重新激活SLG功能的新型治疗靶标。