scRNA-Seq reveals anti-fibrotic mechanisms of TRPC6 inhibition [Spatial Transcriptomics]. scRNA-Seq reveals anti-fibrotic mechanisms of TRPC6 inhibition [Spatial Transcriptomics]

Inhibition of transient receptor potential canonical 6 channels (TRPC6) alleviates tubular injury and renal fibrosis induced by unilateral ureteral obstruction (UUO).However, the exact renoprotective mechanisms are unknown. In this study, we utilized single-cell RNA sequencing (scRNA-Seq) to define the cellular and transcriptional landscape associated with renoprotection through in vivo TRPC6 inhibition, specifically using SH045 (larixyl N-methylcarbamate), in the UUO model. Overall design: Six C57BL/6 mice underwent unilateral ureteral obstruction (UUO) surgery. Seven days post-surgery, the kidneys from Vehicle-treated (n=3) and SH045-treated (n=3) mice were harvested. The contralateral kidneys from Vehicle-treated mice (n=3) served as the control group (Ctrl group). A total of nine kidney samples were used for single-cell RNA sequencing (scRNA-seq). Additionally, two of these samples (one from the SH045 group and one from the Vehicle group) were simultaneously analyzed using spatial transcriptomics.

抑制瞬时受体电位阳离子通道蛋白6（TRPC6）可减轻单侧输尿管梗阻（UUO）诱导的肾小管损伤与肾纤维化，但其确切的肾脏保护机制尚未阐明。本研究以单侧输尿管梗阻模型为研究对象，采用SH045（落叶松脂素N-甲基氨基甲酸酯）实现体内TRPC6抑制，通过单细胞RNA测序（scRNA-Seq）解析其介导肾脏保护的细胞及转录组全貌。整体实验设计如下：选取6只C57BL/6小鼠实施单侧输尿管梗阻手术。术后第7天，分别采集溶剂处理组（n=3）与SH045处理组（n=3）小鼠的肾脏组织；另取溶剂处理组小鼠的对侧肾脏（n=3）作为对照组（Ctrl组）。本研究共纳入9份肾脏样本用于单细胞RNA测序（scRNA-seq）。此外，还选取其中2份样本（分别来自SH045处理组与溶剂处理组）同步开展空间转录组学分析。