Transcriptional profiling of Wilms tumor via single nuclei RNA sequencing

Wilms tumor, or nephroblastoma, is the most common pediatric kidney cancer, and is known to arise from renal precursor cells in the fetal kidney. Given that single nuclei RNA-sequencing generates gene expression profiles of diverse cell types within a heterogeneous tissue, here we sought to utilize this technology to profile blastema, stroma, and epithelial components of favorable histology Wilms tumor samples. Overall design: Frozen, unfixed samples were biobanked at the time of surgical resection/nephrectomy. Nuclei isolation was performed for 10x genomics single nuclei RNA-seq, targeting 5000 nuclei with 50K reads/nuclei from each sample.

肾母细胞瘤（Wilms tumor，nephroblastoma）是儿童最常见的肾脏恶性肿瘤，其起源为胎儿肾脏内的肾前体细胞。鉴于单细胞核RNA测序（single nuclei RNA-sequencing）可获取异质性组织内多种细胞类型的基因表达谱，本研究拟利用该技术对组织学类型良好的肾母细胞瘤样本的胚基（blastema）、间质（stroma）及上皮（epithelial）组分进行转录组谱分析。
实验设计概述：本研究所用样本为手术切除（肾切除术）时留存的冷冻未固定组织，已完成生物样本库入库。针对所有样本，我们均开展细胞核分离操作，用于10x Genomics（10x基因组学平台）单细胞核RNA测序，每个样本的靶向捕获目标为5000个细胞核，测序数据量设置为每个细胞核约5万条读段。