Endocannabinoid system as a therapeutic target for PVR

摘要 两种眼细胞类型，视网膜色素上皮(RPE)和Müler胶质细胞(MG)被认为与其有关 在增殖性玻璃体视网膜病变(PVR)的发展和最终转归中的重要作用 转分化为肌成纤维细胞。内源性大麻素系统，包括内源性大麻素配体和 它们的受体，包括CB1，CB2和非CB1/CB2大麻素受体，在健康和 疾病，是很有希望的治疗靶点。此前，已经表明，封锁CB1和 激活CB2抑制肝纤维化。我们的初步结果表明，肌成纤维细胞的转分化 N-油酰多巴胺(OLDA)是一种内源性反向激动剂，可抑制RPE和MG细胞 GPR6，一个非CB1/CB2的大麻素受体。此外，CB1选择性反向激动剂/拮抗剂SR141716A 抑制MG细胞的肌成纤维细胞转分化。根据文献和我们的初步数据，我们 假设CB1和GPR6反向激动剂/拮抗剂和CB2激动剂抑制肌成纤维细胞改变 通过分别通过CB1、GPR6、CB2受体发挥作用，改变下游信号通路至关重要 用于肌成纤维细胞的转分化。为了验证我们的假设，CB1、CB2和GPR6将被激活 选择性激动剂或被反向激动剂或shRNA敲除抑制以检测它们对肌成纤维细胞的作用 通过间充质和肌成纤维细胞标志物蛋白表达和基质评估转分化 收缩，肌成纤维细胞的关键功能。此外，CB1、CB2和GPR6配体在信号转导中的作用 由促纤维化细胞因子转化生长因子-2激活的通路将通过评估Key的激活状态来检测 信号分子，以及纤维化转录因子的核定位。最后，CB1的表达， CB2和GPR6受体将在PVR患者的人视网膜瘢痕组织中进行研究。的主要目标是 本项目旨在测试CB1、CB2和GPR6配体作为PVR预防性治疗的潜力。 此外，该项目将确定内源性大麻素系统靶向的纤维化信号通路和 应有助于开发针对PVR的特异和有效的治疗药物。

ABSTRACT Two ocular cell types, retinal pigment epithelium (RPE) and Müller glia (MG) have been implicated to play important roles in the development and final outcome of proliferative vitreoretinopathy (PVR) by undergoing Trans differentiation to myofibroblasts. The endocannabinoid system, including endocannabinoid ligands and their receptors, including CB1, CB2, and non-CB1/CB2 cannabinoid receptors, play essential roles in health and disease, and are promising therapeutic targets. Previously, it has been shown that blockade of CB1 and activation of CB2 inhibit fibrosis. Our preliminary results demonstrate that myofibroblast trans differentiation of both RPE and MG cells can be inhibited by N-oleoyl dopamine (OLDA), an endogenous inverse agonist for GPR6, a non-CB1/CB2 cannabinoid receptor. In addition, CB1 selective inverse agonist/antagonist SR141716A inhibited myofibroblast trans differentiation of MG cells. Based on the literature and our preliminary data, we hypothesize that CB1 and GPR6 inverse agonists/antagonists and CB2 agonists inhibit myofibroblastic changes by working via CB1, GPR6, CB2 receptors respectively and modifying downstream signaling pathways crucial for myofibroblast trans differentiation. To test our hypothesis, CB1, CB2 and GPR6 will either be activated by selective agonists or inhibited by inverse agonists or shRNA knockdown to examine their role on myofibroblast trans differentiation, assessed by mesenchymal and myofibroblast marker protein expression and matrix contraction, a key function of myofibroblasts. In addition, effects of CB1, CB2 and GPR6 ligands on signaling pathways activated by profibrotic cytokine TGF2 will be examined by assessing activation status of key signaling molecules, as well as nuclear localization of fibrotic transcription factors. Finally, the expression of CB1, CB2 and GPR6 receptors will be investigated in human retinal scar tissues from PVR patients. The main goal of this project is to test the potential of CB1, CB2 and GPR6 ligands as preventative treatments of PVR. Furthermore, this project will identify fibrotic signaling pathways targeted by the endocannabinoid system and should contribute to the development of specific and effective therapeutic agents for PVR.