Adenosine receptor 2A in subretinal fibrosis

腺苷受体2A在视网膜下纤维化中的作用

• 批准号: 10614638
• 负责人: Ruth B Caldwell
• 金额: $ 43.78万
• 依托单位: AUGUSTA UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-05-01 至 2027-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10614638
• 关键词: Actins; Adenosine; Anatomy; Angioblast; Blood Vessels; Bone Marrow; Cells; Choroidal Neovascularization; Cicatrix; Data; Development; Endothelial Cells; Endothelium; Epithelial Cells; Exudative age-related macular degeneration; Eye diseases; Fibrosis; Genetic; Hypoxia Inducible Factor; Hypoxia-Inducible Factor Pathway; In Vitro; Infiltration; Inflammatory; Lasers; Lesion; Macrophage; Mediating; Mesenchymal; Molecular; Mus; Myelogenous; Myeloid Cells; Myofibroblast; Neuroglia; Oxygen; Pathologic; Patients; Prevention; Production; Profibrotic signal; Purinergic P1 Receptors; Receptor Signaling; Retina; Retinal Diseases; Role; Signal Transduction; Smooth Muscle; Snails; Structure of retinal pigment epithelium; Testing; Therapeutic; Transforming Growth Factor Beta 2; Vascular Diseases; Vascular Endothelial Growth Factors; Visual impairment; cell type; design; in vitro Model; in vivo; inhibitor; mouse model; novel strategies; novel therapeutic intervention; overexpression; pharmacologic; retinal angiogenesis; tool; transcription factor; vascular inflammation

PROJECT SUMMARY Subretinal fibrosis, an end-stage fibrous scar of neovascular age-related macular degeneration (nAMD), compromises highly organized anatomical layers and tightly coordinated cellular interactions, inevitably leading to irreversible visual impairment. The current treatment for subretinal fibrosis is limited and therefore, new therapeutic strategies for the inhibition of subretinal fibrosis are imperative. Multiple cell types, including endothelial cells (ECs), retinal pigment epithelium (RPE) cells, macrophages and glial cells, contribute to subretinal fibrosis by either differentiating into mesenchymal-like cells and further differentiating into α-smooth muscle actin-positive myofibroblasts and/or producing profibrotic and proinflammatory factors. However, the underlying mechanisms for these cellular and molecular activities remain poorly defined. Adenosine receptor 2A (Adora2a) has been implicated in various vascular diseases and inflammation. Our preliminary data here show that (i) the level of Adora2a expression was increased in subretinal lesions of laser-induced CNV in mice; (ii) the size of subretinal fibrosis was markedly decreased in lesions of laser–induced CNV in Adora2a-deficient mice; (iii) endothelial-to-mesenchymal transition (EndMT) occurred to choroidal ECs (CECs) and EndMT participated in the formation of subretinal fibrosis in laser-induced mouse CNV; (iv) Tgfb2-induced EndMT was decreased for Adora2a-deficient CECs; (v) macrophage-to-myofibroblast transition (MMT) in laser-induced subretinal fibrotic lesions was markedly reduced in Adora2a-deficient mice; (vi) Adora2a-deficient bone marrow derived macrophages (BMDMs) had a compromised production of profibrotic factors after stimulation with Tgfb2; and (vii) the levels of hypoxia-inducible factor (Hif) 1a or 2a dynamically correlated with those of Adora2a in the above pathological alterations. Thus, we hypothesize that Adora2a- mediated Hif signaling in CECs and infiltrated macrophages enhance fibrotic effects leading to increased formation of fibrotic lesions in CNV. To test our hypothesis, we have generated mice with inducible global Adora2a deficiency in Vldlr-/- mice, endothelial lineage tracing mice, inducible endothelial Adora2a deficiency in C57BL/6j mice, and myeloid Adora2a deficiency in C57BL/6j mice. We established an ex vivo approach to culture mouse CECs and in vitro approaches to generate BMDMs. We will investigate the effect of Adora2a inactivation in CECs and myeloid cells in subretinal fibrosis using specific genetic and pharmacological tools and assess subretinal fibrosis using an integrated approach of in vivo, ex vivo, and in vitro models. Our study will define the role of Adora2a in the development of subretinal fibrosis and provide the basis for using ADORA2A inhibition as a novel approach in the prevention and treatment of blinding retinal disease.

项目概要 视网膜下纤维化，新生血管性年龄相关性黄斑变性（nAMD）的终末期纤维疤痕， 损害高度组织的解剖层和紧密协调的细胞相互作用，不可避免地导致 目前对视网膜下纤维化的治疗是有限的，因此是新的。 抑制视网膜下纤维化的治疗策略势在必行。 多种细胞类型，包括内皮细胞 (EC)、视网膜色素上皮 (RPE) 细胞、巨噬细胞 和神经胶质细胞，通过分化为间充质样细胞并进一步分化而导致视网膜下纤维化 分化为α-平滑肌肌动蛋白阳性肌成纤维细胞和/或产生促纤维化和 然而，这些细胞和分子活动的潜在机制仍然存在。 腺苷受体 2A (Adora2a) 与多种血管疾病有关。 我们的初步数据表明 (i) Adora2a 表达水平在视网膜下增加。 小鼠激光诱导的 CNV 损伤；(ii) 视网膜下纤维化损伤的大小显着减小。 Adora2a 缺陷小鼠中激光诱导的 CNV；(iii) 发生内皮间质转化 (EndMT)； 脉络膜EC（CEC）和EndMT参与激光诱导小鼠视网膜下纤维化的形成 CNV；(iv) Adora2a 缺陷的 CEC 中 Tgfb2 诱导的 EndMT 降低；(v) 巨噬细胞至肌成纤维细胞 Adora2a 缺陷小鼠中激光诱导的视网膜下纤维化病变的转变 (MMT) 显着减少 (vi) Adora2a 缺陷的骨髓源性巨噬细胞 (BMDM) 的促纤维化产生受损 Tgfb2 刺激后的因子；以及 (vii) 动态缺氧诱导因子 (Hif) 1a 或 2a 的水平 与上述病理改变中的 Adora2a 相关，因此，我们认为 Adora2a-。 CEC 和浸润巨噬细胞中介导的 Hif 信号增强纤维化效应，导致纤维化效应增加 为了检验我们的假设，我们培育了具有诱导性整体的小鼠。 Vldlr-/- 小鼠中的 Adora2a 缺陷，内皮谱系追踪小鼠，诱导型内皮 Adora2a 缺陷 C57BL/6j 小鼠和 C57BL/6j 小鼠的骨髓 Adora2a 缺陷 我们建立了离体培养方法。 小鼠 CEC 和体外生成 BMDM 的方法我们将研究 Adora2a 失活的影响。 使用特定的遗传和药理学工具对视网膜下纤维化中的 CEC 和骨髓细胞进行研究并评估 我们的研究将使用体内、离体和体外模型的综合方法来定义视网膜下纤维化。 Adora2a 在视网膜下纤维化发展中的作用，并为使用 ADORA2A 抑制作用作为基础 一种预防和治疗致盲性视网膜疾病的新方法。