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Abstract: SA-PO612

Spatial Transcriptomics Provides Unique Insights Into the Pathophysiology of Experimental Pyelonephritis

Session Information

  • Pediatric Nephrology - II
    November 05, 2022 | Location: Exhibit Hall, Orange County Convention Center‚ West Building
    Abstract Time: 10:00 AM - 12:00 PM

Category: Pediatric Nephrology

  • 1800 Pediatric Nephrology

Authors

  • Ballash, Gregory, Kidney and Urinary Tract Center, Abigail Wexner Research Institute, Nationwide Children's Hospital, Columbus, Ohio, United States
  • Cotzomi Ortega, Israel, Kidney and Urinary Tract Center, Abigail Wexner Research Institute, Nationwide Children's Hospital, Columbus, Ohio, United States
  • Jackson, Ashley R., Kidney and Urinary Tract Center, Abigail Wexner Research Institute, Nationwide Children's Hospital, Columbus, Ohio, United States
  • Becknell, Brian, Kidney and Urinary Tract Center, Abigail Wexner Research Institute, Nationwide Children's Hospital, Columbus, Ohio, United States
  • Ruiz-Rosado, Juan de Dios, Kidney and Urinary Tract Center, Abigail Wexner Research Institute, Nationwide Children's Hospital, Columbus, Ohio, United States

Group or Team Name

  • Kidney and Urinary Tract Center
Background

Acute pyelonephritis (APN) is considered one of the most significant bacterial infections among young children and is associated with acute kidney injury, renal scarring, and end-stage kidney disease. The molecular mechanisms leading to alterations of tissue homeostasis and long-term sequelae during APN are poorly understood. In this study, we utilized a novel technique, spatially resolved transcriptomics, to define the molecular basis of APN pathology.

Methods

Immunocompetent C3H/HeOuJ female mice underwent transurethral inoculation with uropathogenic Escherichia coli (UPEC). We performed spatial transcriptomics (Visium, 10x Genomics) in kidney sections from infected mice at 0, 7 and 28 days post-infection (dpi), to provide a spatiotemporal context of differential gene expression during UPEC-induced pyelonephritis. Transcriptomic data was spatially integrated using the 10X Space Ranger pipeline. Space Ranger output was investigated for gene expression within tissues via Loupe Browser and across tissue using the Seurat package in R.

Results

APN kidneys showed unique clustering that localized to geographic regions of the cortex and papilla corresponding to renal abscessation. The acute phase of infection witnessed the emergence of four unique clusters associated with immune response initiation, LPS tolerance, and promoting apoptosis. As infection progressed to a chronic phase, two unique clusters localized to early renal scars that prominently featured genes involved in the adaptive immune response, fibroblast migration, and extracellular matrix formation. By integrating single cell RNA-seq analysis, we identified an enrichment of neutrophils in areas of abscessation, surrounded by macrophages at 7 dpi, and an enrichment of stromal cells in regions of mononuclear inflammation at 28 dpi. Predicted pathways of inflammation thought to instigate renal fibrosis circumscribed renal lesions at both acute and chronic phases of infection.

Conclusion

Spatial transcriptomics is a powerful tool to superimpose transcriptional changes onto the landscape of renal pathology. This experimental approach will serve to integrate transcriptional dynamics into a pathophysiological map of the renal response to bacterial infections.

Funding

  • NIDDK Support