Kidney Week

Abstract: FR-PO601

Cell State Transition Associated with Cyst Development in Autosomal Dominant Polycystic Kidney Disease

Session Information

• Cystic Kidney Diseases: Mechanisms and Models
  October 25, 2024 | Location: Exhibit Hall, Convention Center
  Abstract Time: 10:00 AM - 12:00 PM

Category: Genetic Diseases of the Kidneys

• 1201 Genetic Diseases of the Kidneys: Cystic

Authors

• Jung, Hyun Jun, The Johns Hopkins University School of Medicine, Baltimore, Maryland, United States

Hyun Jun Jung, PhD

• Outeda, Patricia, University of Maryland School of Medicine, Baltimore, Maryland, United States

Patricia Outeda, PhD

• Woodward, Owen M., University of Maryland School of Medicine, Baltimore, Maryland, United States

Owen M. Woodward, PhD

• Watnick, Terry J., University of Maryland School of Medicine, Baltimore, Maryland, United States

Terry J. Watnick, MD

• Welling, Paul A., The Johns Hopkins University School of Medicine, Baltimore, Maryland, United States

Paul A. Welling, MD

Background

Autosomal Dominant Polycystic Kidney Disease (ADPKD), caused by genetic mutations in the Polycystin genes (Pkd1 and Pkd2), is the most common form of polycystic kidney disease, characterized by the development of cysts in the kidneys. Mechanisms underlying the initiation of cystogenesis are poorly understood, but developmental stage plays an important role. In mouse models, cyst formation is greatly accelerated when Pkd1/2 genes are knocked out before postnatal day 13 (P13), leading severe cyst development within weeks, compared to months when Pkd1/2 knockout is induced after P13. Here, we investigated the molecular signature of cell state transitions around P13 underlying the dynamic changes in the initiation of cyst development.

Methods

To identify transcriptomic signatures and their changes associated with cell state transitions around P13, single nucleus RNA sequencing (snRNA-Seq) was performed on whole kidneys obtained from mice with an intact Pkd2 gene (Pkd2^fl/fl; Pax8-rtTA; TetO-Cre-) at P13 and P15. Nuclei (n=2 per group, 54,324 nuclei) isolated from whole kidneys were analyzed using 10x Chromium and NovaSeq6000 (50,000 read pairs per nucleus). Data were processed and analyzed using CellRanger and Seurat.

Results

snRNA-Seq identified kidney cell populations according to known kidney cell types. At both P13 and P15, a cell proliferation signature was observed in subpopulations of proximal tubule, loop of Henle, and distal tubule cells. Differential gene expression analysis (P13 vs P15) identified 142 genes commonly changed in proliferative tubular cells and revealed that these genes were enriched in calcium signaling. Further analysis of proliferative tubular cell subclusters identified 6 common genes (A330015K06Rik, Dapk2, Exoc3l2, Gli2, Greb1, and Slc14a2) and revealed that the cell proliferation signature was less prominent at P15 compared to P13. Additionally, 48 genes encoding secretory proteins exhibited cell type-selective changes within proliferative cell subpopulations.

Conclusion

We identified a transcriptomic signature of a proliferative cell state and their changes between P13 and P15. This study offers a valuable resource for understanding cell state transitions in proliferative epithelial cells, potentially implicated in the development of cystogenesis in ADPKD.

Funding

• NIDDK Support