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会导致明显的视觉丧失。使用多种免疫技术，我们评估了这些疾病中免疫细胞的各种特征，包括信号通路，例如炎症和自身免疫途径。 我们已经看到葡萄膜炎的分子特征有所不同。 特别有趣的是IL-22的识别。 IL-22的表达最近与Th17细胞有关，Th17细胞在AMD和葡萄膜炎患者的血清中升高。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的-bad水平。 此外，我们看到与年龄相关的黄斑变性患者的免疫细胞中IL-17活性增加。 我们报告了AMD患者的免疫细胞的表观遗传改变，但这尚待验证。但是，注意到了AMD眼中IL-17及其受体等免疫标记物的存在，无论其机制如何。 此外，类固醇难治性葡萄膜炎的患者具有外周血液中类固醇难治性CD4+ T细胞的特征性亚群。先前的研究表明，这种类固醇难治性表型仅限于具有产生IL-17的CD4+细胞的中央记忆库。因此，我们比较了Th1和Th17细胞对皮质类固醇的转录组反应，以鉴定新型的生物标志物和靶标在类固醇难治性疾病中的治疗干预措施。类固醇难治性患者的倾向比敏感患者更大，但与TH1细胞相比，暴露于DEX后，来自两组患者的Th17细胞的基因表达变化类似限制。 使用其他技术，我们已经确定葡萄膜炎患者的亚组显着缩短了端粒长度。此外，我们注意到与活性疾病有关的结节病患者循环IL-17。我们已经与马里兰大学合作，研究了识别疾病表型的新方法。一种特别有趣的方法是SuperCell Statistics，这是一种基于单元的平均过程，然后是机器学习方案。