增生性玻璃体视网膜病变(PVR)是手术修复的孔源性视网膜脱离(RRD)失败的最常见原因。化学诱导和细胞注射PVR模型不能完全模拟RRD后环境中PVR的临床特征。对研究RRD-PVR特异性机制和治疗的转化模型存在未满足的需求。
■在成年荷兰束带兔中诱导RRD。在诱导后6小时和2、7、14和35天固定或处理后节段用于RNA测序。胶质纤维酸性蛋白的组织化学染色和免疫标记,α平滑肌肌动蛋白,进行血管内皮生长因子受体2、CD68和RPE65kDa蛋白,并对标记强度进行评分。进行单细胞RNA测序。
■急性组织病理学改变包括玻璃体内和视网膜内出血,白细胞性玻璃体炎,脉络膜视网膜炎,和视网膜稀疏。慢性病变表现为视网膜萎缩,胶质增生,纤维化视网膜下膜,视网膜上纤维血管增生.慢性病变的纤维细胞膜和纤维血管膜中存在原纤维胶原。在慢性病变中检测到神经胶质和脉管系统的中度至强标记。在第14天,通过单细胞测序分析的大多数细胞被鉴定为穆勒胶质细胞和小胶质细胞,与免疫标记一致。慢性病变中几种纤维状胶原基因的表达上调。
■该兔子模型的组织学和转录特征模拟了人类RRD-PVR的重要特征,包括向慢性视网膜内和视网膜周围纤维化的过渡。这种具有PVR特征的RRD动物模型将有助于进一步研究靶向治疗干预措施。
UNASSIGNED: Proliferative vitreoretinopathy (PVR) is the most common cause of failure of surgically repaired rhegmatogenous retinal detachment (RRD). Chemically induced and cell injection PVR models do not fully simulate the clinical characteristics of PVR in the post-RRD context. There is an unmet need for translational models in which to study mechanisms and treatments specific to RRD-PVR.
UNASSIGNED: RRD was induced in adult Dutch Belted rabbits. Posterior segments were fixed or processed for RNA sequencing at 6 hours and 2, 7, 14, and 35 days after induction. Histochemical staining and immunolabeling for glial fibrillary acidic protein, alpha smooth muscle actin, vascular endothelial growth factor receptor 2, CD68, and RPE 65 kDa protein were performed, and labeling intensity was scored. Single cell RNA sequencing was performed.
UNASSIGNED: Acute histopathological changes included intravitreal and intraretinal hemorrhage, leukocytic vitritis, chorioretinitis, and retinal rarefaction. Chronic lesions showed retinal atrophy, gliosis, fibrotic subretinal membranes, and epiretinal fibrovascular proliferation. Fibrillar collagen was present in the fibrocellular and fibrovascular membranes in chronic lesions. Moderate to strong labeling of glia and vasculature was detected in chronic lesions. At day 14, most cells profiled by single cell sequencing were identified as Mϋller glia and microglia, consistent with immunolabeling. Expression of several fibrillar collagen genes was upregulated in chronic lesions.
UNASSIGNED: Histological and transcriptional features of this rabbit model simulate important features of human RRD-PVR, including the transition to chronic intraretinal and periretinal fibrosis. This animal model of RRD with features of PVR will enable further research on targeted treatment interventions.