Effect of SS-associated cytokines on salivary gland dysfunction

SS相关细胞因子对唾液腺功能障碍的影响

基本信息

批准号：
7788432
负责人：
Olga Juliana Baker
金额：
$ 22.97万
依托单位：
STATE UNIVERSITY OF NEW YORK AT BUFFALO
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-06-04 至 2012-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7788432
关键词：
Acinus organ component Affect Agonist Animal Model Anions Autoimmune Diseases C10 Calcium Carbachol Cell physiology Cells Cholinergic Receptors Chronic Complex Cytosol Data Down-Regulation Dryness Epithelial Epithelial Cells Excision Figs - dietary Functional disorder Generations Gland In Vitro Individual Inflammation Inflammatory Interferons Interleukin-12 Interleukin-18 Interleukin-6 Intestines Ions Lacrimal gland structure Major salivary gland structure Mediating Messenger RNA Minor salivary gland structure Morphology Mus Muscarinics Oral cavity Parotid Gland Patients Permeability Phosphorylation Plasma Production Protein Biosynthesis Proteins Public Health Publishing Rattus Recovery Resistance Role Saliva Salivary Salivary Gland Diseases Salivary Glands Signal Pathway Signal Transduction Sjogren&apos s Syndrome Staging Structure Tight Junctions Tumor Necrosis Factor Activation Tumor Necrosis Factor-alpha Tumor Necrosis Factors Wild Type Mouse airway epithelium apical membrane base claudin-1 protein cytokine in vivo in vivo Model monolayer mouse model nucleotide receptor occludin overexpression protein distribution protein expression public health relevance receptor-mediated signaling saliva secretion

项目摘要

DESCRIPTION (provided by applicant): Sjogren's syndrome (SS) is an autoimmune disorder characterized by inflammation and dysfunction of salivary glands, resulting in impaired secretory function. Levels of the pro-inflammatory cytokines tumor necrosis factor-1 (TNF1), interferon-3 (IFN3), IL-12, interleukin-6 (IL-6), interleukin-18 (IL-18) and interleukin-12 (IL-12) are elevated in salivary glands of patients with SS, although little is known about the effects of these cytokines on salivary epithelial cell tight junction (TJ) integrity which is necessary to establish transepithelial ion gradients that drive saliva secretion. We have demonstrated that chronic exposure of polarized rat parotid gland (Par-C10) epithelial cell monolayers to TNF1 and/or IFN3 decreases transepithelial resistance (TER) and transepithelial anion secretion induced by carbachol, a muscarinic cholinergic receptor agonist, or UTP, a P2Y2 nucleotide receptor agonist. In contrast, TNF1 and/or IFN3 had no effect on agonist-induced increases in the intracellular calcium concentration [Ca2+]i in Par-C10 cells indicating that individual cell signaling is unaffected by cytokines. Our studies show that among the TJs, claudin-1 is selectively downregulated by TNF1 and/or IFN3. In cells treated with TNF1, claudin-1 downregulation returns to normal levels upon incubation of cells for 24 h in cytokine- free medium. Under these conditions, recovery of claudin-1 expression corresponds with increases in TER and agonist-induced short circuit current (Isc). We have obtained new preliminary data demonstrating that silencing of claudin-1 expression decreases TER in Par-C10 monolayers. Furthermore, the cellular distribution of the TJ proteins occludin and ZO-1 was altered by TNF1 and/or IFN3 treatment of Par- C10 three-dimensional (3D) acinar spheres. Based on these preliminary findings, we will examine the overall hypothesis that cytokine- induced changes in the expression and/or distribution of TJ proteins affects TJ integrity in Par-C10 cell monolayers, consistent with a loss in saliva secretion associated with cytokine generation in the intact parotid gland. In addition, we propose to identify the cellular mechanisms underlying cytokine-induced disruption of TJ integrity using in vitro and in vivo models of SS. Studies in Specific Aim 1 will characterize the mechanisms underlying cytokine-induced decreases in claudin-1 expression. Studies in Specific Aim 2 will identify the TNF1- and IFN3-mediated changes in TJ protein phosphorylation and removal from the TJ complex in Par-C10 cells. Studies in Specific Aim 3 will characterize TJ mRNA and protein expression, TJ morphology, and TJ cellular distribution in parotid glands of the C57BL/6.NOD-Aec1Aec2 mouse model of SS and mice overexpressing TNF1, as compared to wild type mice. These studies will provide a greater understanding the mechanisms whereby saliva secretion is decreased during the pro-inflammatory stages of salivary gland diseases, a significant initial step in defining the poorly-studied signaling pathways that regulate TJ structural integrity in parotid acini. PUBLIC HEALTH RELEVANCE: Sjogren's syndrome (SS) is an autoimmune disease characterized by salivary gland dysfunction leading to severe dryness of the oral cavity. Pro-inflammatory cytokines are up-regulated in plasma and in salivary glands from patients with SS, however, little is known of their role in salivary gland dysfunction. We propose to determine the role of pro-inflammatory cytokines in salivary gland tight junction integrity causing reduction of saliva secretion.

描述（申请人提供）：干燥综合征（SS）是一种自身免疫性疾病，其特征是唾液腺炎症和功能障碍，导致分泌功能受损。促炎细胞因子肿瘤坏死因子 1 (TNF1)、干扰素 3 (IFN3)、IL-12、白细胞介素 6 (IL-6)、白细胞介素 18 (IL-18) 和白细胞介素 12 (IL) 的水平-12) 在 SS 患者的唾液腺中升高，尽管人们对这些细胞因子对唾液上皮细胞紧密连接 (TJ) 完整性的影响知之甚少，而建立唾液上皮细胞紧密连接 (TJ) 完整性是建立必要的条件驱动唾液分泌的跨上皮离子梯度。我们已经证明，极化的大鼠腮腺（Par-C10）单层上皮细胞长期暴露于 TNF1 和/或 IFN3 会降低跨上皮阻力（TER）和由卡巴胆碱（一种毒蕈碱胆碱能受体激动剂）或 UTP（一种 P2Y2）诱导的跨上皮阴离子分泌。核苷酸受体激动剂。相反，TNF1和/或IFN3对Par-C10细胞中激动剂诱导的细胞内钙浓度[Ca2+]i的增加没有影响，这表明个体细胞信号传导不受细胞因子的影响。我们的研究表明，在 TJ 中，claudin-1 被 TNF1 和/或 IFN3 选择性下调。在用 TNF1 处理的细胞中，在无细胞因子的培养基中孵育细胞 24 小时后，claudin-1 下调恢复至正常水平。在这些条件下，claudin-1 表达的恢复与 TER 和激动剂诱导的短路电流 (Isc) 的增加相对应。我们获得了新的初步数据，表明沉默 claudin-1 表达会降低 Par-C10 单层中的 TER。此外，TJ蛋白occludin和ZO-1的细胞分布因Par-C10三维(3D)腺泡球的TNF1和/或IFN3处理而改变。基于这些初步发现，我们将检验总体假设，即细胞因子诱导的 TJ 蛋白表达和/或分布变化会影响 Par-C10 细胞单层中的 TJ 完整性，这与唾液分泌减少与细胞因子生成相关。完整的腮腺。此外，我们建议使用 SS 的体外和体内模型来确定细胞因子诱导的 TJ 完整性破坏的细胞机制。具体目标 1 中的研究将描述细胞因子诱导的claudin-1 表达减少的机制。具体目标 2 中的研究将确定 Par-C10 细胞中 TJ 蛋白磷酸化和 TJ 复合物去除中 TNF1 和 IFN3 介导的变化。具体目标 3 中的研究将表征与野生型小鼠相比，SS 的 C57BL/6.NOD-Aec1Aec2 小鼠模型和过度表达 TNF1 的小鼠腮腺中的 TJ mRNA 和蛋白质表达、TJ 形态和 TJ 细胞分布。这些研究将有助于更好地了解唾液腺疾病促炎阶段唾液分泌减少的机制，这是确定调节腮腺腺泡 TJ 结构完整性的信号通路的重要第一步，这一信号通路研究较少。公共卫生相关性：干燥综合征 (SS) 是一种自身免疫性疾病，其特征是唾液腺功能障碍，导致口腔严重干燥。 SS 患者血浆和唾液腺中促炎细胞因子的表达上调，但人们对其在唾液腺功能障碍中的作用知之甚少。我们建议确定促炎细胞因子在唾液腺紧密连接完整性中的作用，从而导致唾液分泌减少。