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

眼部炎症性疾病，包括葡萄膜炎和黄斑变性，会导致严重的视力丧失。利用各种免疫技术，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。近年来，IL-22的表达与Th17细胞有关，而Th17细胞在AMD和葡萄膜炎患者的血清中升高。我们已经证明，IL-22导致原代培养的RPE细胞凋亡，可能是通过降低磷酸化的Bad水平。此外，我们还观察到老年性黄斑变性患者免疫细胞中IL-17活性升高。 我们已经报道了AMD患者免疫细胞的表观遗传学改变，但这一点仍有待证实。然而，无论其机制是什么，AMD患者眼睛中免疫标志物如IL-17及其受体的存在都有所增加。此外，类固醇难治性葡萄膜炎患者外周血中有一种特有的类固醇难治性CD4+T细胞亚群。以前的研究已经证明，这种类固醇耐药表型仅限于具有产生IL-17能力的中央记忆池的CD4+细胞。因此，我们比较了Th1和Th17细胞对糖皮质激素的转录反应，以确定新的生物标志物和激素难治性疾病治疗干预的靶点。激素难治性患者比敏感患者更容易产生Th17细胞，但与Th1细胞相比，接触地塞米松后，两组患者的Th17细胞在基因表达方面的变化都受到类似的限制。 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，我们还注意到结节病患者循环中的IL-17与活动性疾病有关。我们与马里兰大学合作，寻找识别疾病表型的新方法。一种特别有趣的方法是超级细胞统计学，这是一种基于单个细胞的平均过程，然后是机器学习方案。

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.