ASN's Mission

To create a world without kidney diseases, the ASN Alliance for Kidney Health elevates care by educating and informing, driving breakthroughs and innovation, and advocating for policies that create transformative changes in kidney medicine throughout the world.

learn more

Contact ASN

1401 H St, NW, Ste 900, Washington, DC 20005

email@asn-online.org

202-640-4660

The Latest on X

Kidney Week

Abstract: SA-PO847

Oxford Kidney Pathology Atlas: Single Cellular Profiling of Renal Biopsy Tissue

Session Information

Category: Glomerular Diseases

  • 1401 Glomerular Diseases: From Inflammation to Fibrosis

Authors

  • Kepple, Jessica, University of Oxford, Oxford, United Kingdom
  • Davis, Simon, University of Oxford, Oxford, United Kingdom
  • Fischer, Roman, University of Oxford, Oxford, United Kingdom
  • Bull, Katherine R., University of Oxford, Oxford, United Kingdom

Group or Team Name

  • Oxford Renal Pathology Research Group-Bull.
Background

Glomerular damage is central to many kidney diseases, thus a better understanding of glomerular cell pathology is essential for developing more targeted therapeutics. Transcriptomic and proteomic profiling of human renal tissue may address this challenge. Many studies utilize nephrectomy sections, with limited samples, technical glomerular dissociation bias, and non-standardized methodology restricting the research application of biopsies. As part of the Oxford Kidney Pathology Atlas study, we developed experimental protocols for processing standard human renal biopsy samples for single-nuclei RNA sequencing (snRNA-Seq) and glomerular proteomics and generated pilot data from healthy donors and an early primary idiopathic nephrotic syndrome (iNS) case.

Methods

Biopsy cores from diseased patients and pre-transplantation controls were processed for clinical histology, snRNA-Seq and targeted glomerular proteomics. SnRNA-Seq libraries were analyzed using the R package Seurat. Glomerular regions from formalin-fixed paraffin-embedded (FFPE) renal biopsy sections were isolated via targeted laser capture microdissection coupled with mass spectrometry (MS). MS results were analyzed using DI-ANN and Perseus software to assess relative protein abundance.

Results

We conducted a targeted proteomics titration experiment with varying numbers of isolated glomeruli from healthy donors to establish the minimum material required for proteomics. We isolated glomerular proteins from FFPE-embedded renal section, quantifying over 4.5k unique proteins from a single glomerulus and over 5k proteins from 6-10 human glomeruli. By snRNA-Seq, libraries were generated per single biopsy core, with glomerular and tubular populations represented. Despite normal histology, iNS patient-derived nuclei enriched in select proximal tubule and glomerular cell clusters, with altered gene signatures including increased cellular transport function and extracellular matrix accumulation, and reduced podocyte function.

Conclusion

We demonstrate that glomerular proteomic markers can be captured from 6 glomeruli from a human FFPE biopsy section. Key renal cell types are detectable by snRNA-Seq from standard clinical biopsy core, sufficient to detect differential cell composition and gene expression in disease tissue. This platform will be applied to identify novel therapeutic targets in kidney disease.

Funding

  • Commercial Support – Novo Nordisk