权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Role of lacrimal gland myoepithelial cells in dry eye disease

泪腺肌上皮细胞在干眼病中的作用

基本信息

批准号：
10553199
负责人：
DRISS ZOUKHRI
金额：
$ 48.38万
依托单位：
TUFTS UNIVERSITY BOSTON
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-02-01 至 2025-01-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10553199
关键词：
Acinar Cell Acinus organ component Actins Age Animal Model Atrophic Autoimmune Candidate Disease Gene Catabolism Cell Separation Cell physiology Cells Chronic Contractile Proteins Contracts Coupling Data Denervation Disease Down-Regulation Dry Eye Syndromes Ductal Epithelial Cell Efferent Neurons G-Protein-Coupled Receptors GTP-Binding Proteins Gene Expression Genes Gland Goals Health Human Impairment In Vitro Inflammation Inflammatory Interleukin-1 Knock-out Knowledge Lacrimal gland structure Lead MAPK8 gene MMP2 gene Mammary gland Mediating Metalloproteases Microfilaments Molecular Mus Muscle Muscle Proteins Muscle denervation procedure Muscular Atrophy Myoepithelial cell N-terminal Neuropeptides Output Oxytocin Oxytocin Receptor Pathogenesis Pathway interactions Phosphotransferases Process Production Proteins Proteomics RNA Receptor Cell Receptor Signaling Reporting Role Second Messenger Systems Signal Transduction Signal Transduction Pathway Sjogren&apos s Syndrome Smooth Muscle Smooth Muscle Myocytes Sorting Stress System Techniques Tissues Translating Ubiquitin aqueous calponin cell type cytokine mammary milk production milk secretion multicatalytic endopeptidase complex muscular system neural neurotransmitter release new therapeutic target novel protein expression receptor receptor expression response sex transcriptome sequencing

项目摘要

Chronic inflammation of the lacrimal gland (LG), as occurs in Sjögren’s syndrome, is the leading cause of aqueous- deficient dry eye disease (DED). The mechanisms leading to insufficient LG secretion are still not completely understood. The LG is composed of acinar, myoepithelial, and ductal cells, with acini and myoepithelial cells (MEC) forming the secretory units. MECs express several muscle proteins, such as alpha smooth muscle actin (SMA) and calponin, and are therefore able to contract. MECs are best studied in the mammary gland where their contraction is shown to be crucial for milk production and contraction is mainly controlled by the neuropeptide oxytocin. Despite their potential critical role in LG secretion, very little is known about MEC contraction in this tissue, nor is the impact of chronic inflammation of the LG on these cells. One of goals of the current proposal is to fill this gap in knowledge. Our preliminary studies show that murine and human LG MECs express the oxytocin receptor (OXTR) and contract in response to oxytocin stimulation. Furthermore, we show that MECs in chronically inflamed LG are atrophied, with down-regulation of contractile proteins SMA and calponin and the OXTR and MECs from these glands do not contract in response to oxytocin stimulation. We previously reported that interleukin-1 (IL-1) inhibits neurotransmitter release from LG efferent nerves leading to DED. We also reported activation of the stress activated c-Jun N-terminal kinase (JNK) and metalloproteinases 2 (MMP2) in chronically inflamed LGs and that inhibition of either pathway restored LG secretion and tears output in animal models of DED. Numerous studies showed that denervation of muscle tissues leads to tissue atrophy and degradation of muscle contractile proteins via the ubiquitin/proteasome pathways. Based on these findings, we hypothesize that in chronic DED, proinflammatory cytokines inhibit neurotransmitter release from LG efferent nerves creating a denervated-like tissue and that they trigger degradation of the OXTR and MEC myofilament proteins. This degradation translates into a loss of contractibility of MECs thus further exacerbating the effect of the loss of neural input on the secretory units of the LG. We further hypothesize that the JNK, MMP2, and ubiquitin/proteasome pathways mediate the effects of proinflammatory cytokines on MEC functions. We will use both in vitro (sorted MEC cells) as well as animal models of DED to investigate how proinflammatory cytokines interfere with oxytocin-induced contraction of LG MECs. We will use unbiased RNA-seq and quantitative global proteomics techniques to identify novel pathways that are altered in MECs in chronically inflamed LGs. At the completion of these studies, we will have established a role for the MEC, an important and yet understudied cell in the LG, and the oxytocin signaling system in the pathogenesis of aqueous-deficient DED and identified potential novel therapeutic targets.

泪腺（LG）的慢性炎症，如干燥综合征，是水囊炎的主要原因。缺乏性干眼症（DED）。导致LG分泌不足的机制仍不完全清楚。 LG由腺泡、肌上皮和导管细胞组成，腺泡和肌上皮细胞（MEC）形成分泌单位。MEC表达几种肌肉蛋白，如α平滑肌肌动蛋白（SMA）和钙调蛋白，因此能够收缩。在乳腺中对MEC进行了最好的研究，其中它们的收缩被证明对产奶至关重要，并且收缩主要由神经肽催产素控制。尽管它们在LG分泌中具有潜在的关键作用，但对该组织中MEC收缩的了解甚少，LG的慢性炎症对这些细胞的影响也是如此。目前提案的目标之一是填补这一知识空白。我们的初步研究表明，小鼠和人类LG MEC表达催产素受体（OXTR）和收缩催产素刺激的反应。此外，我们表明，在慢性发炎的LG中的MEC是萎缩的，伴随着收缩蛋白SMA和钙调蛋白的下调，并且来自这些腺体的OXTR和MEC不响应于催产素刺激而收缩。我们以前报道，白细胞介素-1（IL-1）抑制神经递质释放从LG传出神经导致DED。我们还报道了应激激活的c-Jun N-末端激酶（JNK）和金属蛋白酶2（MMP 2）在慢性炎症的LG中的激活，以及在DED动物模型中抑制任一途径恢复LG分泌和泪液分泌。大量研究表明，肌肉组织的去神经支配导致组织萎缩和肌肉收缩蛋白的降解通过泛素/蛋白酶体途径。基于这些发现，我们假设在慢性DED中，促炎细胞因子抑制LG传出神经的神经递质释放，产生去神经支配样组织，并且它们触发OXTR和MEC肌丝蛋白的降解。这种降解转化为一个损失的MEC的收缩性，从而进一步加剧了神经输入的损失对LG的分泌单位的影响。我们进一步假设，JNK，MMP 2和泛素/蛋白酶体途径介导的促炎细胞因子对MEC功能的影响。我们将使用体外（分选的MEC细胞）以及DED的动物模型来研究促炎细胞因子如何干扰催产素诱导的LG MEC收缩。我们将使用无偏见的RNA-seq和定量的全球蛋白质组学技术来确定在慢性炎症的LG中MEC中改变的新途径。在这些研究完成后，我们将建立MEC的作用，一个重要的，但研究不足的细胞在LG，催产素信号系统在缺水性DED的发病机制，并确定潜在的新的治疗靶点。