Endocannabinoid system as a therapeutic target for PVR

内源性大麻素系统作为 PVR 的治疗靶点

• 批准号: 10747004
• 负责人: ZHAO-HUI SONG
• 金额: $ 44.53万
• 依托单位: UNIVERSITY OF LOUISVILLE
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-30 至 2025-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10747004
• 关键词: Agonist; Blindness; CNR1 gene; CNR2 gene; Cannabinoids; Cannabis; Categories; Cells; Chemicals; Cicatrix; Complication; Data; Development; Disease; Dopamine; Endocannabinoids; Extracellular Matrix; Eye Injuries; Failure; Family suidae; Fibrosis; Future; G-Protein-Coupled Receptors; GPR6 gene; Goals; Health; Human; Immunohistochemistry; Inflammation; Investigation; Legal Status; Ligands; Literature; Mediating; Medical Marijuana; Medicine; Mesenchymal; Modeling; Modification; Molecular; Muller' s cell; Myofibroblast; Nuclear; Operative Surgical Procedures; Outcome; Pathology; Patients; Pharmaceutical Preparations; Play; Prevention; Preventive treatment; Proliferative Vitreoretinopathy; Reporting; Research; Retina; Retinal Detachment; Role; SR 141716A; Signal Pathway; Signaling Molecule; Structure of retinal pigment epithelium; Surface; Testing; Therapeutic Agents; Therapeutic Effect; Tissues; Traction; Visual; Visual Acuity; antagonist; antiproliferative drugs; cannabinoid receptor; cell type; design; endogenous cannabinoid system; improved; in vivo; interest; knock-down; mouse model; phytocannabinoid; prevent; profibrotic cytokine; protein biomarkers; protein expression; receptor; response; small hairpin RNA; synthetic cannabinoid; therapeutic target; therapeutically effective; transcription factor; transdifferentiation

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.

抽象的 两种眼细胞类型：视网膜色素上皮细胞 (RPE) 和米勒神经胶质细胞 (MG) 与此有关 增殖性玻璃体视网膜病变（PVR）的发展和最终结果中的重要作用 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，并且是有希望的治疗靶点 此前，已经证明 CB1 和 CB1 的阻断。 我们的初步结果表明，CB2 的激活可抑制纤维化。 RPE 和 MG 细胞均可被 N-油酰多巴胺 (OLDA) 抑制，OLDA 是一种内源性反向激动剂 GPR6，一种非 CB1/CB2 大麻素受体，另外，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。 坚持认为 CB1 和 GPR6 反向激动剂/拮抗剂和 CB2 激动剂抑制肌成纤维细胞变化 分别通过 CB1、GPR6、CB2 受体起作用并修改至关重要的下游信号通路 为了检验我们的假设，CB1、CB2 和 GPR6 将被激活。 选择性激动剂或通过反向激动剂或 shRNA 敲低抑制，以检查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记蛋白表达和基质评估转分化 收缩是肌成纤维细胞的一个关键功能。此外，CB1、CB2 和 GPR6 配体对信号传导的影响。 将通过评估关键因子的激活状态来检查促纤维化细胞因子 TGF2 激活的途径 信号分子以及纤维化转录因子的核定位最后是 CB1 的表达。 CB2 和 GPR6 受体将在 PVR 患者的人类视网膜疤痕组织中进行研究。 该项目旨在测试 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 的特异性且有效的治疗药物。