The role of protein O-GlcNAcylation in regulating keratinocyte function in skin fibrosis

蛋白O-GlcNAc酰化在调节皮肤纤维化角质形成细胞功能中的作用

基本信息

批准号：
10725270
负责人：
YAN WANG
金额：
$ 40.14万
依托单位：
CLEVELAND CLINIC LERNER COM-CWRU
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-09-23 至 2025-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10725270
关键词：
Adipose tissue Affect Alzheimer&apos s Disease Atrophic condition of skin Biological Assay Bleomycin Blindness Chemicals Child Chronic Disease Collagen Contracture Data Defect Deposition Development Diabetes Mellitus Disease End Point Assay Epidermis Excision Extracellular Matrix Fascia Fibroblasts Fibrosis Formalin Future Gene Silencing Growth Growth Factor Hair Harvest Human In Vitro Incidence Induction of Apoptosis Inflammatory Investigation Joints Knowledge Lead Localized scleroderma Malignant Neoplasms Mass Spectrum Analysis Methods Modeling Morphea Mus Muscle Myofibroblast Names O-GlcNAc transferase Paraffin Embedding Pathogenesis Pathology Pathway interactions Patients Persons Phase Production Profibrotic signal Protein Inhibition Proteins Proteome Proteomics Role Serine Severities Site Skin Skin Abnormalities Skin Tissue Subcutaneous Tissue Systemic Scleroderma Testing Threonine Tissues Uracil Nucleotides Uridine Diphosphate N-Acetylglucosamine bone bone loss cytokine effective therapy genetic regulatory protein in vivo inhibitor insight keratinocyte laser capture microdissection mouse model novel novel therapeutic intervention paracrine peptide O-linked N-acetylglucosamine-beta-N-acetylglucosaminidase pharmacologic profibrotic cytokine response skin fibrosis skin organogenesis subcutaneous transcriptome sequencing transcriptomics treatment strategy

项目摘要

Project Summary Localized scleroderma (LS), also named morphea, is a sclerotic disorder that affects not only skin, but also adjacent tissues including subcutaneous adipose tissue, fascia, muscle, and bone. LS can cause devastating permanent defects including hair loss, atrophy of skin, subcutaneous tissue and bone loss, joint contractures, vision loss, and growth restriction of affected body sites in children. The disease mechanism of LS is poorly understood and current treatments are unsatisfactory. Better understanding of the disease and more effective treatment strategies are clearly needed. LS and systemic sclerosis (SSc) share the same histopathologic features, and have two phases: an early inflammatory stage and a later fibrotic stage. Upregulated proinflammatory and profibrotic signals in the skin lead to hyperactivation of fibroblasts, which is the cellular hallmark for both LS and SSc and results in excessive collagen production and increased deposition of extracellular matrix (ECM). Epidermal keratinocytes, by secreting various cytokines, have a crucial role in regulating fibroblast function via paracrine effects. Currently, a significant gap in knowledge is how the paracrine effects between keratinocytes and fibroblasts are initiated and regulated. Protein O-GlcNAcylation is the addition of the GlcNAc moiety from nucleotide uridine diphosphate-N-acetyl-glucosamine (UDP-GlcNAc) onto serine or threonine residues of cytosolic proteins. This is catalyzed by O-GlcNAc transferase (OGT), and GlcNAc is removed by O-GlcNAcase (OGA). Abnormal O-GlcNAcylation of proteins is found in various chronic diseases including diabetes, cancers, and Alzheimer’s disease. However, the role of protein O-GlcNAcylation in the pathogenesis of LS has not been studied. We propose to test the hypothesis that abnormal levels of intracellular protein O-GlcNAcylation in skin lead to altered production of proinflammatory and profibrotic cytokines by keratinocytes, which in turn promotes fibroblast activation via paracrine effects. Inhibition of protein O-GlcNAcylation may provide a novel therapeutic strategy to alleviate the abnormal skin fibrosis pathology in LS and SSc. Aim 1: Determine the role of protein O-GlcNAcylation in regulating the production of proinflammatory and profibrotic cytokines by keratinocytes. Aim 2: Determine the paracrine regulatory effects of keratinocytes with altered protein O-GlcNAcylation on fibroblast function and the development of fibrosis in vivo. We expect to show that: 1) Manipulation of protein O-GlcNAcylation in keratinocytes will affect their production of key proinflammatory and profibrotic cytokines; 2) changes in keratinocyte function will affect fibroblast activation and profibrotic functions; and 3) Treatment with chemical inhibitors to OGT or OGA will affect the development of skin fibrosis induced by Bleomycin. Impact: Positive results from our proposed study will provide strong justification for further investigation of this idea and provide novel insights into the mechanisms by which keratinocytes regulate activation of fibroblasts in dysregulated fibrosis pathologies such as LS from a new glycobiological perspective (protein O-GlcNAcylation) that has been significantly overlooked.

项目概要局限性硬皮病 (LS)，也称为硬斑病，是一种硬化性疾病，不仅影响皮肤，还影响皮肤邻近组织包括皮下脂肪组织、筋膜、肌肉和骨骼，可引起毁灭性的 LS。永久性缺陷，包括脱发、皮肤萎缩、皮下组织和骨质流失、关节挛缩、儿童视力丧失和受影响身体部位的生长受限 LS 的发病机制尚不明确。对这种疾病的了解和目前的治疗方法并不令人满意。 LS 和系统性硬化症 (SSc) 显然需要相同的组织病理学特征。具有两个阶段：早期炎症阶段和晚期纤维化阶段。皮肤中的促炎和促纤维化信号导致成纤维细胞过度活化，这是细胞的 LS 和 SSc 的标志，导致胶原蛋白过度生成和胶原蛋白沉积增加细胞外基质（ECM）通过分泌各种细胞因子，在表皮角质形成细胞中发挥着至关重要的作用。通过旁分泌作用调节成纤维细胞功能目前，人们对如何通过旁分泌作用调节成纤维细胞功能存在重大认识空白。角质形成细胞和成纤维细胞之间的旁分泌作用被启动和调节。添加来自核苷酸尿苷二磷酸-N-乙酰基-葡萄糖胺 (UDP-GlcNAc) 的 GlcNAc 部分到胞浆蛋白的丝氨酸或苏氨酸残基上，这是由 O-GlcNAc 转移酶 (OGT) 催化的，并且 GlcNAc 被 O-GlcNAcase (OGA) 去除，蛋白质的异常 O-GlcNAc 酰化存在于各种慢性疾病中。然而，蛋白质 O-GlcNAc 酰化的作用。我们建议检验 LS 发病机制中异常水平的假设。皮肤中细胞内蛋白 O-GlcNAc 酰化导致促炎和促纤维化物质的产生改变角质细胞产生细胞因子，进而通过旁分泌作用的抑制促进成纤维细胞活化。蛋白O-GlcNAc酰化可能为缓解异常皮肤纤维化提供新的治疗策略 LS 和 SSc 的病理学目标 1：确定蛋白质 O-GlcNAc 酰化在调节产生的作用中的作用。目的 2：确定角质形成细胞的旁分泌调节作用。蛋白 O-GlcNAc 酰化改变的角质形成细胞对成纤维细胞功能和纤维化发展的影响我们期望证明：1) 角质形成细胞中蛋白质 O-GlcNAc 酰化的操作将影响其关键促炎和促纤维化细胞因子的产生；2）角质形成细胞功能的变化会影响成纤维细胞活化和促纤维化功能；3) 使用 OGT 或 OGA 化学抑制剂治疗将影响博来霉素诱导的皮肤纤维化的发展。影响：我们提出的研究的积极结果。将为进一步研究这一想法提供强有力的理由，并提供新的见解角质形成细胞在失调的纤维化病理中调节成纤维细胞活化的机制，例如从新的糖生物学角度（蛋白质 O-GlcNAcylation）来看，LS 已被严重忽视。