Adenosine receptor 2A in subretinal fibrosis

腺苷受体2A在视网膜下纤维化中的作用

基本信息

批准号：
10417359
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
项目类别：
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/10417359
关键词：
Anatomy Angioblast Blood Vessels Bone Marrow Cells Choroidal Neovascularization Cicatrix Data Development Endothelial Cells Endothelium Exudative age-related macular degeneration Eye diseases Fibrosis Genetic Hypoxia Inducible Factor Hypoxia-Inducible Factor Pathway In Vitro Lasers Lesion Mediating Mesenchymal Molecular Mus Myelogenous Myeloid Cells Myofibroblast Neuroglia Oxygen Pathologic Patients Pharmacology Prevention Production Profibrotic signal Purinergic P1 Receptors Receptor Signaling Retina Retinal Diseases Role Signal Transduction Smooth Muscle Actin Staining Method Structure of retinal pigment epithelium Testing Therapeutic Vascular Diseases Vascular Endothelial Growth Factors Visual impairment cell type design in vitro Model in vivo inhibitor macrophage mouse model novel strategies novel therapeutic intervention overexpression retinal angiogenesis tool transcription factor vascular inflammation

项目摘要

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.

项目摘要视网膜下纤维化，与新血管相关的黄斑变性（NAMD）的终末期纤维疤痕，妥协高度组织的解剖层和紧密协调的细胞相互作用，不可避免地领导不可逆的视觉障碍。当前对视视纤维化的治疗受到限制，因此，抑制视网膜下纤维化的治疗策略必须进行。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，通过区分间充质样细胞并进一步促进肾上腺素纤维化区分α平滑肌肌动蛋白阳性肌纤维细胞和/或产生纤维化和/或促炎性因素。但是，这些细胞和分子活性的基本机制仍然存在定义不佳。腺苷受体2a（adora2a）已在各种血管疾病中隐含炎。我们的初步数据表明（i）视网膜下adora2a表达的水平增加小鼠激光诱导的CNV病变；（ii）视网膜下纤维化的大小明显降低激光诱导的adora2a缺陷小鼠中的CNV；（iii）内皮到间质转变（ENDMT）发生到脉络膜EC（CEC）和EndMT参与激光诱导的小鼠的视网膜下纤维化的形成 CNV; （iv）针对Adora2a缺乏CECS提高了TGFB2诱导的EndMT；（v）巨噬细胞到肌纤维细胞在Adora2a缺乏小鼠中，激光诱导的视网膜下纤维化病变中的过渡（MMT）显着降低。（vi） Adora2a缺陷骨髓衍生的巨噬细胞（BMDMS）的生产受损用TGFB2刺激后的因素；（vii）动态的缺氧诱导因子（HIF）1a或2a的水平与上述病理改变中的Adora2a相关。那我们假设Adora2a-- CEC中介导的HIF信号传导和浸润的巨噬细胞增强了纤维化作用，导致增加 CNV中纤维化病变的形成。为了检验我们的假设，我们已经产生了具有诱导全局的小鼠 adora2a vldlr-/ - 小鼠缺乏，内皮谱系追踪小鼠，可诱导的内皮adora2a缺乏 C57BL/6J小鼠和C57BL/6J小鼠中的粒细胞Adora2a缺乏。我们建立了一种体内文化方法小鼠CEC和体外方法生成BMDM。我们将研究Adora2a失活的效果在CEC和髓样细胞中，使用特定的遗传和药物工具以及评估在视网膜下纤维化中视网膜下纤维化使用体内，体内和体外模型的整合方法。我们的研究将定义 Adora2a在视网膜下纤维化发展中的作用，为使用adora2a抑制作用提供了基础一种用于预防和治疗盲目残留疾病的新方法。