Yoshida Teruhiko, Latt Khun Zaw, Rosenberg Avi Z, Shrivastav Shashi, Heymann Jurgen, Halushka Marc K, Winkler Cheryl A, Kopp Jeffrey B
Kidney Disease Section, Kidney Diseases Branch, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, Maryland, USA.
Department of Pathology, Johns Hopkins Medical Institutions, Baltimore, Maryland, USA.
Kidney Int Rep. 2022 Oct 17;8(1):164-178. doi: 10.1016/j.ekir.2022.10.011. eCollection 2023 Jan.
The mechanisms in podocytes that mediate the pathologic effects of the high-risk (HR) variants remain incompletely understood, although various molecular and cellular mechanisms have been proposed. We previously established conditionally immortalized human urine-derived podocyte-like epithelial cell (HUPEC) lines to investigate HR variant-induced podocytopathy.
We conducted comprehensive transcriptomic analysis, including mRNA, microRNA (miRNA), and transfer RNA fragments (tRFs), to characterize the transcriptional profiles in undifferentiated and differentiated HUPEC with HR (G1/G2, 2 cell lines) and low-risk (LR) (G0/G0, 2 cell lines) genotypes. We reanalyzed single-cell RNA-seq data from urinary podocytes from focal segmental glomerulosclerosis (FSGS) subjects to characterize the effect of genotypes on podocyte transcriptomes.
Differential expression analysis showed that the ribosomal pathway was one of the most enriched pathways, suggesting that altered function of the translation initiation machinery may contribute to variant-induced podocyte injury. Expression of genes related to the elongation initiation factor 2 pathway was also enriched in the HR urinary podocytes from single-cell RNA-seq, supporting a prior report on the role of this pathway in -associated cell injury. Expression of microRNA and tRFs were analyzed, and the profile of small RNAs differed by both differentiation status and genotype.
We have profiled the transcriptomic landscape of human podocytes, including mRNA, miRNA, and tRF, to characterize the effects of differentiation and of different genotypes. The candidate pathways, miRNAs, and tRFs described here expand understanding of -associated podocytopathies.
尽管已经提出了各种分子和细胞机制，但足细胞中介导高风险（HR）变异体病理效应的机制仍未完全明了。我们之前建立了条件永生化的人尿源足细胞样上皮细胞（HUPEC）系，以研究HR变异体诱导的足细胞病变。
我们进行了全面的转录组分析，包括mRNA、微小RNA（miRNA）和转移RNA片段（tRF），以表征未分化和分化的具有HR（G1/G2，2个细胞系）和低风险（LR）（G0/G0，2个细胞系）基因型的HUPEC中的转录谱。我们重新分析了局灶节段性肾小球硬化（FSGS）患者尿足细胞的单细胞RNA测序数据，以表征基因型对足细胞转录组的影响。
差异表达分析表明核糖体途径是最富集的途径之一，这表明翻译起始机制功能的改变可能导致变异体诱导的足细胞损伤。单细胞RNA测序中，与延伸起始因子2途径相关的基因表达在HR尿足细胞中也有富集，支持了先前关于该途径在相关细胞损伤中作用的报道。我们分析了miRNA和tRF的表达，小RNA的谱因分化状态和基因型而异。
我们描绘了人足细胞的转录组图谱，包括mRNA、miRNA和tRF，以表征分化和不同基因型的影响。这里描述的候选途径、miRNA和tRF扩展了对相关足细胞病变的理解。
