Kidney Week

Abstract: PO0665

Identification of Cell-Specific Transcriptomic Changes and Cross-Talk in Diabetic Mice with Podocyte-Specific Induction of KLF6 Using Single Nuclei RNA Sequencing

Session Information

• Diabetic Kidney Disease: Basic - I
  November 04, 2021 | Location: On-Demand, Virtual Only
  Abstract Time: 10:00 AM - 12:00 PM

Category: Diabetic Kidney Disease

• 601 Diabetic Kidney Disease: Basic

Authors

• Owusu Frimpong, Bismark, Stony Brook University Renaissance School of Medicine, Stony Brook, New York, United States

Bismark Owusu Frimpong

• Gujarati, Nehaben A., Stony Brook University Renaissance School of Medicine, Stony Brook, New York, United States

Nehaben A. Gujarati,

• Mallipattu, Sandeep K., Stony Brook University Renaissance School of Medicine, Stony Brook, New York, United States

Sandeep K. Mallipattu,

Background

Krüppel-Like Factor 6 (KLF6), a zinc finger transcription factor, is a key regulator of cytochrome c oxidase assembly in the podocytes. We previously reported that podocyte-specific loss of KLF6 exacerbates diabetic kidney disease (DKD). However, the role of podocyte-specific induction of human KLF6 in DKD remains unexplored.

Methods

A combination of unilateral nephrectomy and streptozotocin (UNx-STZ) was utilized to induce DKD in mice. Using the “Tet-on” system, mice with podocyte-specific induction of human KLF6 (hKLF6^PODTA) were generated by crossing NPHS2-rtTA mice with tre-hKLF6 mice and fed with doxycycline-containing diet. UNx-STZ-NPHS2-rtTA and SHAM UNx-Veh mice served as diabetic and non-diabetic controls, respectively. Single nucleus (sn)RNA-seq libraries were prepared from kidney cortex using the 10X Chromium System. Raw sequencing data was aligned to mouse pre-mRNA reference genome using Cell Ranger. Quality control, dimensionality reduction and clustering were performed using the R-package, Seurat.

Results

23 clusters were generated using unsupervised clustering analysis. Enrichment and pathway analyses showed a downregulation of injury-related pathways such as inflammatory and interleukin signaling in the UNx-STZ-treated hKLF6^PODTA group across the podocyte, endothelial cell, mesangial cell, and proximal tubular clusters, compared to the UNx-STZ-treated NPHS2-rtTA group. Conversely, metabolic pathways such as oxidative phosphorylation and fatty acid metabolism were upregulated. A cross-reference of the differentially expressed genes (DEGs) in the podocyte cluster of the UNx-STZ-treated hKLF6^PODTA group with a KLF6 ChIP-seq data set revealed the presence of putative KLF6 binding sites in the regulatory regions of several DEGs. A unique proximal tubule (PT) cluster with distinctive gene expression signature was identified in the hKLF6^PODTA group, suggesting an intercellular communication between podocytes and the PT cells in the hKLF6^PODTA group that mediates the progression of DKD.

Conclusion

SnRNA-seq demonstrates potential mechanisms by which podocyte-specific induction of KLF6 attenuates the progression of DKD.

Funding

• NIDDK Support