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（RPE）和Müller胶质（mg）已牵涉到 通过进行增生的玻璃体炎（PVR）的重要作用在开发和最终结果中 与肌纤维细胞的跨分化。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，是有希望的治疗靶标。以前，已经表明CB1和 CB2抑制纤维化的激活。我们的初步结果表明，肌纤维细胞的跨分化 RPE和MG细胞均可通过N-烯酰胺多巴胺（Olda）抑制，这是一种内源性反向激动剂 GPR6，一种非CB1/CB2大麻素受体。此外，CB1选择性逆激动剂/拮抗剂SR141716A 抑制MG细胞的肌纤维细胞跨分化。根据文献和我们的初步数据，我们 假设CB1和GPR6反激动剂/拮抗剂和CB2激动剂抑制肌纤维细胞的变化 通过分别通过CB1，GPR6，CB2受体工作并修改下游信号通路至关重要 用于肌纤维细胞跨分化。为了检验我们的假设，CB1，CB2和GPR6要么被激活 选择性激动剂或被反向激动剂或shRNA敲低抑制，以检查其在肌纤维细胞上的作用 通过间充质和肌纤维细胞标记蛋白表达和基质评估的跨分化 收缩，肌纤维细胞的关键功能。此外，CB1，CB2和GPR6配体对信号的影响 通过评估钥匙的激活状态，将检查由纤维化细胞因子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，CB1的表达 CB2和GPR6受体将在PVR患者的人类残留疤痕组织中进行研究。主要目标 该项目是为了测试CB1，CB2和GPR6配体作为PVR的预防性处理的潜力。 此外，该项目将确定内源性大麻素系统和 应为PVR的特定有效治疗剂的开发做出贡献。