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

• 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
• 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' 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

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.

项目摘要 局限性硬皮病（LS），也称为硬斑病，是一种不仅影响皮肤， 邻近组织，包括皮下脂肪组织、筋膜、肌肉和骨。LS可以造成毁灭性的 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， 视力下降和儿童受影响身体部位的生长受限。LS的发病机制尚不清楚 目前的治疗方法并不令人满意。更好地了解疾病，更有效地 治疗策略显然是必要的。LS和系统性硬化症（SSc）具有相同的组织病理学 特征，并具有两个阶段：早期炎症阶段和后期纤维化阶段。上调 皮肤中的促炎和促纤维化信号导致成纤维细胞的过度活化，成纤维细胞是皮肤中的细胞增殖因子。 这是LS和SSc的标志，并导致过度的胶原蛋白产生和增加的胶原蛋白沉积。 细胞外基质（ECM）。表皮角质形成细胞通过分泌各种细胞因子，在细胞增殖中起关键作用。 通过旁分泌作用调节成纤维细胞功能。目前，知识的一个重大差距是， 启动并调节角质形成细胞和成纤维细胞之间的旁分泌作用。蛋白质O-GlcNAc化是 添加来自核苷酸尿苷二磷酸-N-乙酰基-葡糖胺（UDP-GlcNAc）的GlcNAc部分 连接到胞质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， GlcNAc被O-GlcNAc酶（OGA）去除。蛋白质的异常O-GlcNAc化在各种慢性炎症中被发现。 包括糖尿病、癌症和阿尔茨海默病在内的疾病。然而，蛋白质O-GlcNAc化的作用 在LS发病机制中的作用尚未研究。我们建议测试的假设，异常水平的 皮肤中细胞内蛋白O-GlcNAc酰化导致促炎和促纤维化产生改变 角质形成细胞分泌细胞因子，这反过来又通过旁分泌效应促进成纤维细胞活化。抑制 蛋白O-GlcNAc化可能为减轻异常皮肤纤维化提供一种新的治疗策略 LS和SSc中的病理学。目的1：确定蛋白质O-GlcNAc化在调节蛋白质合成中的作用。 促炎和促纤维化细胞因子。目的2：确定旁分泌调节效应 蛋白质O-GlcNAc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 vivo.我们期望表明：1）角质形成细胞中蛋白质O-GlcNAc化的操纵将影响其细胞增殖。 产生关键的促炎和促纤维化细胞因子; 2）角质形成细胞功能的变化将影响 成纤维细胞活化和促纤维化功能;和3）用OGT或OGA的化学抑制剂治疗将 影响博莱霉素诱导的皮肤纤维化的发展。影响：我们拟议研究的积极结果 将提供强有力的理由，进一步调查这一想法，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， 作为LS从一个新的糖生物学的角度（蛋白质O-GlcNAc酰化），已被显着忽视。