Age-Related Meibomian Gland Dysfunction

年龄相关的睑板腺功能障碍

• 批准号: 8710230
• 负责人: James V Jester
• 金额: $ 37.85万
• 依托单位: UNIVERSITY OF CALIFORNIA-IRVINE
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-30 至 2015-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8710230
• 关键词: Acinar Cell; Acinus organ component; Adipocytes; Aging; Agonist; Apoptosis; Atrophic; Binding; Cell Culture System; Cell Line; Cell Nucleus; Cell Proliferation; Cells; Cytoplasm; Development; Down-Regulation; Duct (organ) structure; Eye diseases; Eyelid Diseases; Eyelid structure; Gene Expression; Gene Expression Profile; Genes; Gland; Goals; Human; Image; Knowledge; Ligands; Lipids; Measures; Microscopy; Molecular; Mus; Nuclear; Nuclear Receptors; Operative Surgical Procedures; Optics; PPAR gamma; Phosphorylation; Population; Post-Translational Protein Processing; Prevalence; Raman Spectrum Analysis; Recruitment Activity; Regulation; Role; Serine; Simian virus 40; Testing; Tissues; Western Blotting; abstracting; age effect; age related; aging population; base; eye dryness; human NCOR1 protein; in vivo; lipid biosynthesis; meibomian gland; meibomian gland dysfunction; novel; novel therapeutics; promoter; receptor; response; tomography; tool

Project Summary/Abstract The long-term goal of this project is to understand the mechanism of age-related meibomian gland dysfunction (MGD) and evaporative Dry Eye. Recently we have shown that mouse and human meibomian glands undergo specific age-related changes including decreased acinar cell proliferation, acinar atrophy, and altered peroxisome proliferator-activated receptor gamma (PPARg) localization from cytoplasmic-vesicluar/nuclear in young to nuclear in old mice and humans. Since PPARg is a lipid sensitive, nuclear receptor implicated in regulating adipocyte and sebocyte differentiation and lipogenesis, our findings suggest that PPARg may be involved in modulating meibomian gland differentiation during aging. Based on these findings we propose that aging of the meibomian gland may result in down-regulation of PPARg leading to decreased meibocyte differentiation and lipid synthesis, gland atrophy, and a hyposecretory MGD. Currently, there is a MAJOR GAP in knowledge regarding the role of PPARg in meibomian gland function. To test this hypothesis we have develop novel imaging and cell culture systems to assess gland volume, lipid synthesis and their regulation by PPARg. Using non-linear optical (NLO) microscopy and array tomography we have volumetrically reconstructed the mouse meibomian gland and measured total, cellular and lipid volumes in young and old glands. Preliminary studies suggest that atrophy of aging meibomian glands involves a marked loss in the lipid volume suggesting decreased meibocyte differentiation. Additionally, we have used coherent anti-stokes raman spectroscopy (CARS) to identify the regional lipid profiles within the meibomian gland and have tentatively shown that there is an age-related change in the maturation of meibomian gland lipids moving from the acini into the duct. Furthermore, we have developed an SV40 immortalized mouse meibocyte cell line that synthesizes lipids and expresses PPARg. Using these novel tools we propose the following Specific Aims. (1). Establish the age-related changes in PPARg localization and associated gene expression patterns by quantifying the subcellular localization, post-translational modification and downstream response gene expression patterns in the mouse and human meibomian gland. (2) Determine the effects of aging on the meibomian gland by quantifying the volume and lipid synthesis using NLO microscopy and array tomography to volumetrically reconstruct the meibomian gland and CARS to assess regional changes in lipid components present in the acini, ductule and duct of the mouse and human meibomian gland. (3) Assess the effects of natural and synthetic PPARg ligands on lipid synthesis by quantifying the subcellular localization, post- translational modification and downstream response gene expression patterns in cultured mouse meibocytes. (4) Measure the effect of PPARg ligands on meibocyte differentiation in vivo by quantifying the changes in PPARg expression, meibocyte proliferation, gland volume and lipid synthesis in young and old mouse meibomian glands.

项目摘要/摘要 这个项目的长期目标是了解与年龄相关的眉毛腺功能障碍的机制。 (MGD)和蒸发性干眼。最近，我们发现老鼠和人类的眉毛腺都经历了 与年龄相关的特殊变化，包括腺泡细胞增殖减少、腺泡萎缩和改变 过氧化物酶体增殖物激活受体γ(PPARg)在细胞质-囊泡/核中的定位 在老年小鼠和人类中，从年轻到有核。由于PPARg是一种脂类敏感物质，核受体参与了 通过调节脂肪细胞和皮脂细胞的分化和脂肪生成，我们的发现提示PPARg可能是 在衰老过程中参与调节眉板腺的分化。基于这些发现，我们建议 眉板腺的衰老可能导致PPARg的下调，从而导致巨噬细胞的减少 分化和脂肪合成，腺体萎缩，以及记忆减退。目前，有一个 在有关PPARg在眉板腺功能中的作用的知识方面存在重大差距。为了测试这一点 假设我们已经开发了新的成像和细胞培养系统来评估腺体体积、脂质合成 以及PPARg对它们的监管。使用非线性光学(NLO)显微镜和阵列断层扫描 体积重建小鼠眉间腺并测量总体积、细胞体积和脂肪体积 年轻和年老的腺体。初步研究表明，老化的眉板腺的萎缩与显著的 脂肪体积的丧失表明小细胞分化减少。此外，我们还使用了连贯的 反斯托克斯拉曼光谱学(CARS)识别眉板腺内局部脂质分布和 已初步表明，眉板腺脂类的成熟与年龄有关。 从腺泡进入导管。此外，我们还建立了SV40永生化小鼠巨噬细胞系。 合成脂质并表达PPARg。使用这些新工具，我们提出了以下具体目标。 (1)。通过以下方法建立PPARg定位和相关基因表达模式的年龄相关变化 亚细胞定位、翻译后修饰和下游反应基因的定量 在小鼠和人类眉毛腺中的表达模式。(2)确定老化对老年人的影响 用NLO显微镜和阵列断层扫描定量测定眉板腺的体积和脂肪合成 对额肌腺和CARS进行体积重建，以评估局部脂质成分的变化 存在于鼠和人的汗腺的腺泡、导管和导管中。(三)评估工作成效 天然和合成的PPARg配体通过量化亚细胞定位对脂质合成的影响，后 培养的小鼠巨噬细胞的翻译修饰和下游反应基因表达模式。 (4)定量检测PPARg配体在体内对巨噬细胞分化的影响 幼年和老年小鼠PPARG表达、巨噬细胞增殖、腺体体积和脂质合成 眉板腺。