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Abstract: TH-PO808

Spatial Transcriptomic Analysis Reveals Altered Gene Expression in Glomerular Parietal Epithelial Cells Following Tubular Injury

Session Information

Category: Pathology and Lab Medicine

  • 1800 Pathology and Lab Medicine

Authors

  • Yang, Haichun, Vanderbilt University Medical Center, Nashville, Tennessee, United States
  • Zhong, Jianyong, Vanderbilt University Medical Center, Nashville, Tennessee, United States
  • Zhao, Shilin, Vanderbilt University Medical Center, Nashville, Tennessee, United States
  • Kruse, Angela R.S., Vanderbilt University, Nashville, Tennessee, United States
  • Malek, Morad C., Vanderbilt University, Nashville, Tennessee, United States
  • Spraggins, Jeffrey M., Vanderbilt University, Nashville, Tennessee, United States
  • Fogo, Agnes B., Vanderbilt University Medical Center, Nashville, Tennessee, United States
Background

In our previous study, we observed that acute tubular injury leads to the activation and transdifferentiation of parietal epithelial cells (PECs), resulting in a decrease in the ratio of columnar to flat PECs. In this study, our objective was to investigate the spatial transcriptomics of PECs following tubular injury.

Methods

We conducted experiments using wild-type (WT) mice and transgenic mice expressing the diphtheria toxin (DT) receptor in proximal tubular epithelial cells (PTECs). The mice were injected with DT at week 0 and week 1 and sacrificed after 6 weeks. Paraffin-embedded tissue sections were analyzed using the NanoString GeoMx DSP platform.

Results

Using fluorescent antibodies, we successfully isolated glomerular tuft and PECs and performed GeoMx spatial transcriptomics analysis. Principal component analysis (PCA) clearly distinguished tuft and PECs in both DT-treated and normal mice. In DT-treated PECs, we observed increased expression of 153 genes, including osteopontin (OPN), which showed enrichment in pathways related to actin cytoskeleton regulation, apelin signaling, the citric acid cycle, and respiratory electron transport. Immunostaining revealed that osteopontin was undetectable in PECs of WT mice, whereas its expression was significantly increased at week 3 and week 6 after tubular injury in DT-treated mice.

Conclusion

Our findings indicate that proximal tubular injury affects glomerular PECs, leading to their activation and altered gene expression. This study suggests potential pathways involved in the communication between tubular and glomerular compartments.

Funding

  • NIDDK Support