Vascular Remodeling and the Role of PECAM-1 in Sjogren Syndrome

血管重塑和 PECAM-1 在干燥综合征中的作用

• 批准号: 8716273
• 负责人: Andrew Douglas McCall
• 金额: $ 2.81万
• 依托单位: STATE UNIVERSITY OF NEW YORK AT BUFFALO
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-07-01 至 2017-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8716273
• 关键词: Antibodies; Area; Autoimmune Diseases; Binding; Blocking Antibodies; Blood; Blood Vessels; Blood flow; CD31 Antigens; Cell Adhesion Molecules; Cells; Chronic; Co-Immunoprecipitations; Deglutition; Dental caries; Development; Disease; Endothelial Cells; Food; Gland; Goals; Health; Human; Immigration; In Vitro; Infiltration; Inflammation; Inflammatory; Intercellular Fluid; Knowledge; Laboratories; Lead; Lymphocyte; Lymphocytic Infiltrate; Major salivary gland structure; Mass Spectrum Analysis; Mediating; Mediator of activation protein; Microscopy; Minor salivary gland structure; Modeling; Molecular; Mus; Nitric Oxide; Oral health; Organ; Parotid Gland; Pathogenesis; Pathology; Patients; Pattern; Periodontal Diseases; Phenotype; Play; Post-Translational Protein Processing; Process; Protein Isoforms; Quality of life; Research; Reverse Transcriptase Polymerase Chain Reaction; Role; Saliva; Salivary; Salivary Glands; Signal Pathway; Signal Transduction; Sjogren' s Syndrome; Tissues; Tyrosine Phosphorylation Site; Variant; Vascular Endothelial Growth Factors; Vascular remodeling; Western Blotting; Xerostomia; angiogenesis; cadherin 5; in vivo; injured; insight; monolayer; mouse model; novel; response; saliva secretion; tandem mass spectrometry; trafficking

DESCRIPTION (provided by applicant): Saliva plays a critical role in maintaining proper oral health. Hyposalivation can severely reduce the quality of life for an individual.1,2 A common cause of hyposalivation is Sjogren's syndrome (SS), an autoimmune disorder characterized by lymphocytic infiltration into the salivary glands.3 Although many laboratories around the world are investigating SS,4-6 little is known about the role of vascular remodeling (angiogenesis) in the pathogenesis of this disease. Previous studies indicate that mediators of angiogenesis (i.e., vascular endothelial growth factors and nitric oxide) are upregulated in the salivary glands and saliva of SS patients.7,8 These findings suggest that angiogenesis is occurring during the progression of this disorder. Another important contributor to angiogenesis is the cell adhesion protein, platelet endothelial cell adhesion molecule-1 (PECAM-1),9,10 whose expression has not been studied in SS. PECAM-1 can also impact the trafficking of lymphocytes to the salivary gland,11,12 potentially leading to a more severe SS phenotype. The goal of this project is understand how PECAM-1 influences angiogenesis and lymphocyte trafficking in the progression of SS. To conduct this research, we will use human minor salivary glands with SS and a mouse model of SS. Together, the following studies will further our understanding of SS pathogenesis, and may eventually lead to the development of novel therapies for this disorder: Aim 1: To investigate signs of angiogenesis in salivary glands with and without SS. Blood vessel coverage area and expression levels of endothelial markers will be analyzed by fluorescent microscopy, RT-PCR and Western blot. Aim 2: To determine PECAM-1 isoforms, post-translational modifications and signaling pathways in salivary glands with and without SS. PECAM-1 isoforms, post-translational modifications, and signaling partners will be analyzed by RT-PCR, tandem mass spectrometry, and co-immunoprecipitation. Aim 3: To analyze the role of PECAM-1 during lymphocyte transmigration in mouse major salivary glands with SS. PECAM-1 will be functionally blocked in vitro and in vivo using blocking antibodies and transmigration will be quantified.

描述（由申请人提供）：唾液在维持适当的口腔健康方面起着关键作用。唾液分泌减少可严重降低个体的生活质量。1，2唾液分泌减少的常见原因是干燥综合征（SS），这是一种以淋巴细胞浸润到唾液腺为特征的自身免疫性疾病。3尽管世界各地的许多实验室都在研究SS，4-6但对血管重塑（血管生成）在该疾病发病机制中的作用知之甚少。以前的研究表明，血管生成的介质（即，血管内皮生长因子和一氧化氮）在SS患者的唾液腺和唾液中上调。7，8这些发现表明在这种疾病的进展过程中发生了血管生成。血管生成的另一个重要因素是细胞粘附蛋白，血小板内皮细胞粘附分子-1（PECAM-1），9，10，其在SS中的表达尚未研究。PECAM-1还可以影响淋巴细胞向唾液腺的运输，11，12可能导致更严重的SS表型。本项目的目标是了解PECAM-1如何影响SS进展中的血管生成和淋巴细胞运输。为了进行这项研究，我们将使用患有SS的人类小唾液腺和SS的小鼠模型。总之，以下研究将进一步我们的SS发病机制的理解，并可能最终导致这种疾病的新疗法的发展：目的1：调查的迹象，唾液腺血管生成与SS和没有。将通过荧光显微镜、RT-PCR和Western blot分析血管覆盖面积和内皮标志物的表达水平。目的2：研究唾液腺中PECAM-1的亚型、翻译后修饰和信号通路。将通过RT-PCR、串联质谱和免疫共沉淀分析PECAM-1亚型、翻译后修饰和信号传导伴侣。目的3：探讨PECAM-1在SS小鼠大涎腺淋巴细胞迁移中的作用。PECAM-1将在体外和体内使用阻断抗体进行功能性阻断， 要量化。