■增生性玻璃体视网膜病变(PVR)是视网膜复位手术失败的最常见原因,导致这种异常伤口愈合过程的分子变化目前尚不清楚。我们的最终目标是通过使用单细胞转录组学解剖细胞异质性来研究PVR的发病机理。
■在这里,我们旨在比较兔模型中视网膜PVR样品的单细胞RNA测序(scRNA-seq)和单核RNA测序(snRNA-seq)。
■在以对侧眼作为对照的兔眼中单侧诱导PVR损伤。
■在荷兰束带兔中单侧诱发增生性玻璃体视网膜病变。在PVR诱导后的不同时间点,将视网膜解离成细胞或细胞核悬浮液,并处理scRNA-seq或snRNA-seq。
■PVR诱导后视网膜的单细胞和核转录组特征。
■在疾病诱导后4小时和14天对视网膜进行单细胞RNA测序和snRNA-seq。尽管在scRNA-seq样本中独特分子标识符和基因的捕获率更高,单个细胞类型的整体基因表达谱在scRNA-seq和snRNA-seq之间高度相关.两种测序方式之间的主要差异是细胞类型捕获率,然而,神经胶质细胞类型在scRNA-seq中过度表达,通过snRNA-seq富集内部视网膜神经元。此外,纤维化Müller胶质细胞在snRNA-seq样本中过度表达,而反应性Müller胶质细胞在scRNA-seq样品中过度代表。两种方法之间的轨迹分析相似,允许对scRNA-seq和snRNA-seq数据集进行组合分析。
■这些发现突出了scRNA-seq和snRNA-seq分析的局限性,并暗示两种技术一起使用可以比单独使用更准确地识别PVR中异常纤维发生的关键转录网络。
■专有或商业披露可以在参考文献之后找到。
UNASSIGNED: Proliferative vitreoretinopathy (PVR) is the most common cause of failure of retinal reattachment surgery, and the molecular changes leading to this aberrant wound healing process are currently unknown. Our ultimate goal is to study PVR pathogenesis by employing single-cell transcriptomics to dissect cellular heterogeneity.
UNASSIGNED: Here we aimed to compare single-cell RNA sequencing (scRNA-seq) and single-nucleus RNA-sequencing (snRNA-seq) of retinal PVR samples in the rabbit model.
UNASSIGNED: Unilateral induction of PVR lesions in rabbit eyes with contralateral eyes serving as controls.
UNASSIGNED: Proliferative vitreoretinopathy was induced unilaterally in Dutch Belted rabbits. At different timepoints after PVR induction, retinas were dissociated into either cells or nuclei suspension and processed for scRNA-seq or snRNA-seq.
UNASSIGNED: Single cell and nuclei transcriptomic profiles of retinas after PVR induction.
UNASSIGNED: Single-cell RNA sequencing and snRNA-seq were conducted on retinas at 4 hours and 14 days after disease induction. Although the capture rate of unique molecular identifiers and genes were greater in scRNA-seq samples, overall gene expression profiles of individual cell types were highly correlated between scRNA-seq and snRNA-seq. A major disparity between the 2 sequencing modalities was the cell type capture rate, however, with glial cell types overrepresented in scRNA-seq, and inner retinal neurons were enriched by snRNA-seq. Furthermore, fibrotic Müller glia were overrepresented in snRNA-seq samples, whereas reactive Müller glia were overrepresented in scRNA-seq samples. Trajectory analyses were similar between the 2 methods, allowing for the combined analysis of the scRNA-seq and snRNA-seq data sets.
UNASSIGNED: These findings highlight limitations of both scRNA-seq and snRNA-seq analysis and imply that use of both techniques together can more accurately identify transcriptional networks critical for aberrant fibrogenesis in PVR than using either in isolation.
UNASSIGNED: Proprietary or commercial disclosure may be found after the references.