Role of Caveolin-1 in the Maintenance of Blood-retinal Barrier Integrity

项目总结 Caveolin-1(Caveolin-1)，传统上被认为是小窝膜结构域的标志性蛋白，发挥着 在血-视网膜屏障完整性和视网膜炎症中起关键作用。Cav1中的基因变异与 与原发性开角型青光眼和Cav1蛋白在葡萄膜视网膜和 糖尿病视网膜病变模型。我们发现Cav-1和小窝在血-视网膜中起着重要的作用 屏障(BRB)和炎症诱导的细胞因子释放。有趣的是，Cav1仅从 神经视网膜隔区(Müler胶质细胞和神经元)抑制细胞因子释放和免疫细胞内流 在炎症性和神经退行性侮辱之后。当Cav1在几个视网膜炎症中上调时 情况下，我们假设局部操作Cav1功能提供了一种可行的治疗方法 视网膜炎症应激。鉴于目前以类固醇为基础的治疗视网膜炎症性疾病的方法不是 完全有效，并充满潜在的严重副作用，我们假设Cav1代表一个 抑制视网膜炎症的新治疗靶点。因此，理解这一功能是至关重要的。 蛋白质在视网膜应激适应反应的背景下。我们已经做出了令人振奋的观察， 神经视网膜Müler神经胶质细胞中的Cav1不存在于传统的、形态上可识别的 小窝。这是因为分化的Müler神经胶质细胞通常不表达显著水平的Cavin1， 通过与Cav1的相互作用，是形成小窝所必需的。到目前为止，在所有研究的组织中，Cavin1的丢失导致 Cav1蛋白稳定性的一致性丧失。然而，在Müler glia中，Cav1稳定表达(在 在没有Cavin1的情况下)。这提供了一个独特的机会来研究非腔隙的作用。 Cav1与人类疾病有关的器官中的Cav1我们假设这个非空洞的 定位对于Cav1的S促进细胞因子、趋化因子和生长因子的分泌是至关重要的。在……里面 在这项提议中，我们将通过诱导Müler神经胶质细胞表达，在培养和体内测试这一新概念。 洞穴1将洞穴1隔离在洞穴内。我们还将利用异常稳定的表达方式 Cav1，以检查非Caveolae Cav1结构域的组成(在此称为Cav1 支架“)，并鉴定Cav1在没有Cavin1的情况下稳定表达的机制。这些目标有 清除

Project summary Caveolin-1 (Cav1), traditionally viewed as the signature protein of caveolae membrane domains, plays crucial roles in blood-retinal barrier integrity and retinal inflammation. Gene variants in Cav1 are associated with risk of primary open angle glaucoma and Cav1 protein expression is upregulated in uveitic retinas and in models of diabetic retinopathy. We have found that Cav-1 and caveolae play important roles in blood-retinal barrier (BRB) and inflammation-induced cytokine release. Intriguingly, ablation of Cav1 only from the neuroretinal compartment (Müller glia and neurons) suppresses cytokine release and immune cell influx following inflammatory and neurodegenerative insult. As Cav1 is upregulated in several retinal inflammatory conditions, we hypothesize that local manipulation of Cav1 function presents a viable therapy to suppress retinal inflammatory stress. Given that current steroid-based therapies for retinal inflammatory disease are not completely effective and fraught with potentially severe side effects, we hypothesize that Cav1 represents a novel therapeutic target to suppress retinal inflammation. Thus, it is crucial to understand the function of this protein in the context of the retinal stress adaptation response. We have made the exciting observation that Cav1 in Müller glial cells of the neural retina does not reside within traditional, morphologically-identifiable caveolae. This is because differentiated Müller glia do not normally express significant levels of Cavin1 which, via interaction with Cav1, is necessary to form caveolae. In all tissues studied to date, loss of Cavin1 results in coincident loss of Cav1 protein stability. However, in Müller glia, Cav1 is stably expressed (outside of caveolae) in the absence of Cavin1. This provides a unique opportunity to examine the role of non-caveolar Cav1 in an organ in which Cav1 is associated with human disease. We hypothesize that this non-caveolar localization is crucial to Cav1’s ability to promote secretion of cytokines, chemokines, and growth factors. In this proposal, we will test this novel concept in Müller glia in culture and in vivo by inducing the expression of Cavin1 to sequester Cav1 within caveolae. We will also take advantage of the unusually stable expression of Cav1 outside of caveolae to examine the composition of the non-caveolae Cav1 domain (called herein “Cav1 scaffolds”) and to identify the mechanism by which Cav1 is stably expressed without Cavin1. These goals have clear