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.

摘要 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）已被牵连发挥作用， 在增生性玻璃体视网膜病变（PVR）的发展和最终结果中起重要作用， 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CB 1、CB 2和非CB 1/CB 2大麻素受体，在健康和 疾病，是有前途的治疗目标。以前，已经表明阻断CB 1和 CB 2的活化抑制纤维化。我们的初步研究结果表明，肌成纤维细胞转分化， RPE和MG细胞都可以被N-油酰多巴胺（OLDA）抑制，OLDA是一种内源性反向激动剂， GPR 6，一种非CB 1/CB 2大麻素受体。此外，CB 1选择性反向激动剂/拮抗剂SR 141716 A 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CB 1和GPR 6反向激动剂/拮抗剂和CB 2激动剂抑制成肌纤维细胞变化 分别通过CB 1、GPR 6、CB 2受体发挥作用，并修饰下游信号通路， 用于肌成纤维细胞转分化。为了验证我们的假设，CB 1，CB 2和GPR 6将被激活， 选择性激动剂或被反向激动剂或shRNA敲低抑制，以检查它们对肌成纤维细胞的作用 转分化，通过间充质和肌成纤维细胞标志物蛋白表达和基质 收缩，肌成纤维细胞的关键功能。此外，CB 1、CB 2和GPR 6配体对信号传导的影响也被研究。 将通过评估促纤维化细胞因子TGF β 2激活的关键信号通路的激活状态来检查促纤维化细胞因子TGF β 2激活的通路。 信号分子，以及纤维化转录因子的核定位。最后，研究了CB 1， 将在PVR患者的人视网膜瘢痕组织中研究CB 2和GPR 6受体。的主要目标 本项目旨在测试CB 1、CB 2和GPR 6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， 这将有助于开发特异性和有效的PVR治疗药物。