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.

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 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。此外，我们还观察到年龄相关性黄斑变性患者免疫细胞中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，但这仍有待证实。在葡萄膜炎的动物模型中，我们已经看到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观察到变化。此外，免疫标记物如IL-17及其受体的存在增加， 在AMD的眼睛被注意到，无论他们的机制可能是什么。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具有更大的产生Th 17细胞的倾向，并且与Th 1细胞相比，来自任一组患者的Th 17细胞在暴露于Dex后的反应不同。使用基因表达谱，注意到对糖皮质激素的反应受限。有趣的是，有一个大的基因组转移，以回应环孢素A。由于类固醇抗性是一个重要的临床问题，该信息将强烈地表明靶向Th 17细胞或它们所来源的效应记忆T辅助细胞群体的新治疗剂将是药物开发的候选者。令人感兴趣的是发现在人类中，糖皮质激素治疗影响单核细胞的亚型，诱导与T调节细胞的诱导相关的亚型。 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。发现结节病患者在CD 8+细胞上的IL-17 RC表达升高。此外，我们已经注意到，CD 8 + T细胞，现在被指定为Tc 17细胞，产生IL-17，并以CD 146表达为特征。我们还研究了眼部炎症性疾病患者的树突状细胞。我们注意到的是CD 1c +mDCs与活性的相关性，这似乎是由TNF-α-p38 MAPK调节的。我们正在追求的假设，这可能是一个生物标志物的活动之前，疾病成为临床上明显的。