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: 9155561
• 负责人: Robert Nussenblatt
• 金额: $ 25.86万
• 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/9155561
• 关键词: Adrenal Cortex Hormones; Affect; Age related macular degeneration; Animal Model; Apoptosis; Autoimmune Process; Autoimmune Responses; Biological Markers; Blindness; CD4 Positive T Lymphocytes; CD8-Positive T-Lymphocytes; CD8B1 gene; Cells; Characteristics; Clinical; Cyclosporine; DNA; Dendritic Cells; Disease; Epigenetic Process; Exposure to; Eye; Gene Expression; Gene Expression Profiling; Gene Expression Regulation; Genome; Glucocorticoids; Helper-Inducer T-Lymphocyte; Human; Immune; Immunologic Markers; In Vitro; Inflammatory; Inflammatory Response; Interleukin-17; Length; Leukocytes; MAP Kinase Gene; MAPK14 gene; MCAM gene; Memory; Molecular Profiling; Pathway interactions; Patients; Pharmaceutical Preparations; Phenotype; Population; RNA; Refractory; Refractory Disease; Regulatory T-Lymphocyte; Reporting; Sarcoidosis; Serum; Signal Pathway; Steroid Resistance; Steroids; Subgroup; T-Lymphocyte; Techniques; Telomere Shortening; Th1 Cells; Therapeutic Intervention; Tumor Necrosis Factor-alpha; Uveitis; active control; drug development; human TNF protein; interest; interleukin-22; monocyte; novel; novel therapeutics; peripheral blood; receptor; response; 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. In an animal model for uveitis, we have seen that epigenetic altering medication affects positively the expression of active disease, and changes were noted in vitro on human cells. In addition 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, and Th17 cells from either group of patients respond differently following exposure to Dex as compared with Th1 cells. Using gene expression profiling a restricted response to glucocorticoids was noticed. Of interest that there was a large genome shift in response to cyclosporine A. Since steroid resistance is an important clinical problem, this information would suggest strongly that new therapeutic which target either Th17 cells or the effector memory T helper cell population from which they are derived would be candidates for drug development. Of interest is the finding that in humans, glucocorticoid therapy affects the subtype of monocytes, inducing the subtype that is associated with the induction of T regulatory 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. Patients with sarcoidosis were noted to have an elevated IL-17RC expression on CD8+ cells. In addition we have noted that CD8+ T cells, which are now designated as Tc17 cells, produce IL-17 and are characterized by CD146 expression. We have also studies dendritic cells in patients with ocular inflammatory disease. What was noted was the association of CD1c+mDCs with activity, and this appears to be regulated by TNF-alpha-p38 MAPK. We are pursuing the hypothesis that this could be a biomarker for activity before the disease becomes clinically apparent.

眼部炎症性疾病，包括葡萄膜炎和 AMD，会导致严重的视力丧失。使用 通过各种免疫技术，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。我们已经看到了葡萄膜炎的不同分子特征。特别令人感兴趣的是 IL-22 的鉴定。最近发现 IL-22 的表达与 AMD 和葡萄膜炎患者血清中升高的 Th17 细胞有关。我们已经证明，IL-22 可能通过降低磷酸化-Bad 水平导致培养的原代 RPE 细胞凋亡。此外，我们发现年龄相关性黄斑变性患者的免疫细胞中 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，但这仍有待验证。在葡萄膜炎动物模型中，我们发现表观遗传改变药物对活动性疾病的表达产生积极影响，并且在体外人类细胞上也注意到了变化。此外，IL-17 及其受体等免疫标记物的存在也有所增加 AMD 认为，无论其机制是什么。此外，类固醇难治性葡萄膜炎患者的外周血中存在类固醇难治性 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 能力的 CD4+ 细胞的中央记忆池。因此，我们比较了 Th1 和 Th17 细胞对皮质类固醇的转录组反应，以确定类固醇难治性疾病治疗干预的新生物标志物和靶标。类固醇难治性患者比敏感患者更倾向于产生 Th17 细胞，并且与 Th1 细胞相比，两组患者在接触 Dex 后的反应不同。使用基因表达谱分析发现对糖皮质激素的反应有限。有趣的是，对环孢菌素 A 的反应存在较大的基因组转变。由于类固醇耐药性是一个重要的临床问题，这一信息强烈表明，新的治疗方法可以靶向 Th17 细胞或它们所衍生的效应记忆 T 辅助细胞群将成为药物开发的候选者。有趣的是，在人类中，糖皮质激素治疗会影响单核细胞亚型，诱导与 T 调节细胞诱导相关的亚型。 使用其他技术，我们发现葡萄膜炎患者的一个亚组的端粒长度明显缩短。此外，我们注意到结节病患者中循环的 IL-17 与活动性疾病相关。结节病患者的 CD8+ 细胞上 IL-17RC 表达升高。此外，我们还注意到 CD8+ T 细胞（现在被称为 Tc17 细胞）产生 IL-17，并以 CD146 表达为特征。我们还研究了眼部炎症疾病患者的树突状细胞。值得注意的是 CD1c+mDC 与活性的关联，这似乎受到 TNF-α-p38 MAPK 的调节。我们正在追求这样的假设：这可能是疾病在临床上变得明显之前的活动生物标志物。