Renal Endothelial Single-Cell Transcriptomics Reveals Spatiotemporal Regulation and Divergent Roles of Differential Gene Transcription and Alternative Splicing in Murine Diabetic Nephropathy.
Alex-Xianghua ZhouMarie JeanssonLiqun HeLeif WiggePernilla ToneliusRamesh TatiLinda CederbladLars MuhlMartin UhrbomJianping LiuAnna Björnson GranqvistLilach Orly LermanChrister BetsholtzPernille B L HansenPublished in: International journal of molecular sciences (2024)
Endothelial cell (EC) injury is a crucial contributor to the progression of diabetic kidney disease (DKD), but the specific EC populations and mechanisms involved remain elusive. Kidney ECs ( n = 5464) were collected at three timepoints from diabetic BTBR ob/ob mice and non-diabetic littermates. Their heterogeneity, transcriptional changes, and alternative splicing during DKD progression were mapped using SmartSeq2 single-cell RNA sequencing (scRNAseq) and elucidated through pathway, network, and gene ontology enrichment analyses. We identified 13 distinct transcriptional EC phenotypes corresponding to different kidney vessel subtypes, confirmed through in situ hybridization and immunofluorescence. EC subtypes along nephrons displayed extensive zonation related to their functions. Differential gene expression analyses in peritubular and glomerular ECs in DKD underlined the regulation of DKD-relevant pathways including EIF2 signaling, oxidative phosphorylation, and IGF1 signaling. Importantly, this revealed the differential alteration of these pathways between the two EC subtypes and changes during disease progression. Furthermore, glomerular and peritubular ECs also displayed aberrant and dynamic alterations in alternative splicing (AS), which is strongly associated with DNA repair. Strikingly, genes displaying differential transcription or alternative splicing participate in divergent biological processes. Our study reveals the spatiotemporal regulation of gene transcription and AS linked to DKD progression, providing insight into pathomechanisms and clues to novel therapeutic targets for DKD treatment.
Keyphrases
- single cell
- diabetic nephropathy
- rna seq
- gene expression
- transcription factor
- genome wide identification
- dna repair
- genome wide
- type diabetes
- endothelial cells
- high throughput
- copy number
- dna methylation
- wound healing
- dna damage
- high glucose
- genome wide analysis
- dna damage response
- skeletal muscle
- adipose tissue
- heat shock
- metabolic syndrome
- signaling pathway
- insulin resistance
- cell proliferation
- oxidative stress
- genetic diversity
- smoking cessation
- network analysis