Use Of Microarrays and Epigenetics In Gene Expression Of Uveitis & AMD Patients

微阵列和表观遗传学在葡萄膜炎基因表达中的应用

• 批准号: 8938308
• 负责人: Robert Nussenblatt
• 金额: $ 23.03万
• 依托单位: NATIONAL EYE INSTITUTE
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8938308
• 关键词: Adrenal Cortex Hormones; Age related macular degeneration; Apoptosis; Autoimmune Process; Autoimmune Responses; Biological Markers; Blindness; CD4 Positive T Lymphocytes; Cells; Characteristics; DNA; Disease; Epigenetic Process; Exposure to; Eye; Gene Expression; Gene Expression Regulation; Immune; Immunologic Markers; Inflammatory; Inflammatory Response; Interleukin-17; Length; Leukocytes; Machine Learning; Maryland; Memory; Molecular Profiling; Pathway interactions; Patients; Pharmaceutical Preparations; Phenotype; Procedures; RNA; Refractory; Refractory Disease; Reporting; Sarcoidosis; Scheme; Serum; Signal Pathway; Steroids; Subgroup; Techniques; Telomere Shortening; Th1 Cells; Therapeutic Intervention; Universities; Uveitis; active control; base; disease phenotype; interest; interleukin-22; novel; peripheral blood; receptor; response; statistics; transcriptomics

Ocular inflammatory diseases, including uveitis and AMD, cause significant visual loss. Using a variety of immune techniques we have evaluated various characteristics of immune cells in these diseases, including signaling pathways, e.g. inflammatory and autoimmune pathway. We have seen varying molecular signatures for uveitis. Of particular interest is the identification of IL-22. The expression of IL-22 has been recently associated with Th17 cells which is elevated in the serum of AMD and uveitis patients. We have shown that IL-22 resulted in apoptosis in cultured primary RPE cells, possibly by decreasing the phosphorylated-Bad level. In addition, we saw increased IL-17 activity in the immune cells of patients with age related macular degeneration. We have reported epigenetic alterations the immune cells of AMD patients but this remains to be verified. However, an increase in the presence of immune markers such as IL-17 and its receptor in the eyes of AMD were noted, whatever their mechanism may be. In addition, patients with steroid refractory uveitis have a characteristic subpopulation of steroid refractory CD4+ T cells in their peripheral blood. Previously studies have demonstrated that this steroid refractory phenotype is restricted to the central memory pool of CD4+ cells which have the capacity to generate IL-17. We therefore compared transcriptomic responses of Th1 and Th17 cells to corticosteroids in order to identify novel biomarkers and targets for therapeutic intervention in steroid refractory disease. Steroid refractory patients have a greater propensity than sensitive patients to generate Th17 cells, but Th17 cells from either group of patients have a similarly restricted change in gene expression following exposure to Dex compared with Th1 cells. Using additional techniques we have identified that a subgroup of uveitis patients have markedly shortened telomere length. In addition we have noted circulating IL-17 in sarcoidosis patients which is associated with active disease. We have collaborated with the University of Maryland to look at new ways to identify disease phenotypes. One particularly interesting approach has been that of supercell statistics, a single cell based averaging procedure followed by a machine learning scheme.

包括葡萄膜炎和AMD在内的眼部炎症性疾病导致显著的视力丧失。使用各种免疫技术，我们已经评估了这些疾病中免疫细胞的各种特征，包括信号传导途径，例如炎症和自身免疫途径。 我们已经看到葡萄膜炎的不同分子特征。 特别感兴趣的是IL-22的鉴定。IL-22的表达最近与AMD和葡萄膜炎患者血清中升高的Th 17细胞相关。我们已经表明，IL-22导致培养的原代RPE细胞凋亡，可能是通过降低磷酸化Bad水平。 另外我们 在年龄相关性黄斑变性患者的免疫细胞中观察到IL-17活性增加。 我们已经报道了AMD患者免疫细胞的表观遗传改变，但这仍有待证实。然而，注意到AMD患者眼中免疫标记物如IL-17及其受体的存在增加，无论其机制如何。 此外，类固醇难治性葡萄膜炎患者在其外周血中具有类固醇难治性CD 4 + T细胞的特征性亚群。先前的研究已经证明，这种类固醇难治性表型仅限于具有产生IL-17能力的CD 4+细胞的中央记忆库。因此，我们比较了Th 1和Th 17细胞对皮质类固醇的转录组反应，以确定新的生物标志物和类固醇难治性疾病的治疗干预靶点。类固醇难治性患者比敏感性患者更倾向于产生Th 17细胞，但与Th 1细胞相比，来自任一组患者的Th 17细胞在暴露于Dex后的基因表达变化相似地受到限制。 使用额外的技术，我们已经确定，一个亚组的葡萄膜炎患者有显着缩短端粒长度。此外，我们注意到结节病患者的循环IL-17与活动性疾病相关。我们与马里兰州大学合作，寻找识别疾病表型的新方法。一个特别有趣的方法是超级细胞统计，一个基于单个细胞的平均过程，然后是机器学习方案。