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Kidney Week

Abstract: FR-OR118

Regional Transcriptomic Profiling of the Human Kidney Uncovers Major Signature Shifts in the Interstitium During Diabetes

Session Information

Category: Diabetic Kidney Disease

  • 601 Diabetic Kidney Disease: Basic

Authors

  • Barwinska, Daria, Indiana University, Indianapolis, Indiana, United States
  • Cheng, Yinghua, Indiana University School of Medicine, Indianapolis, Indiana, United States
  • Melo ferreira, Ricardo, Indiana University, Indianapolis, Indiana, United States
  • Ferkowicz, Michael J., Indiana University, Indianapolis, Indiana, United States
  • Winfree, Seth, Indiana University School of Medicine, Indianapolis, Indiana, United States
  • Lake, Blue, UCSD, San Diego, California, United States
  • Menon, Rajasree, University of Michigan, Ann Arbor, Michigan, United States
  • Otto, Edgar A., University of Michigan, Ann Arbor, Michigan, United States
  • Collins, Kimberly S., Indiana University School of Medicine, Indianapolis, Indiana, United States
  • Dunn, Ken, Indiana University, Indianapolis, Indiana, United States
  • Kelly, Katherine J., Indiana University, Indianapolis, Indiana, United States
  • Sutton, Timothy A., Indiana University School of Medicine, Indianapolis, Indiana, United States
  • Rovin, Brad H., Ohio State University Wexner Medical Center, Columbus, Ohio, United States
  • Parikh, Samir V., Ohio State University Medical Center, Columbus, Ohio, United States
  • Dagher, Pierre C., Indiana University, Indianapolis, Indiana, United States
  • El-Achkar, Tarek M., Indiana University, Indianapolis, Indiana, United States
  • Eadon, Michael T., Indiana University Division of Nephrology, Indianapolis, Indiana, United States
Background

The expression signature of the renal interstitium is less well characterized than that of the glomerulus and tubules. Here, we examine gene expression of the human renal interstitium in reference nephrectomies and diabetic kidney biopsy specimens using laser micro-dissection (LMD).

Methods

We used LMD to collect cortical kidney interstitium from reference nephrectomies (N=9) and diabetic renal biopies (N=6). Rapid stain with DAPI, OG-Phalloidin, and Tamm-Horsfall protein antibody identified relevant renal structures. LMD excluded tubules, glomeruli and large vessels. Transcriptomic data was obtained using RNAseq on Illumina platform, and analyzed with R studio. The gene signature was compared to existing platforms such as snRNAseq and scRNAseq datasets.

Results

Our laser micro-dissected interstitial regions did not show significant expression of known tubular or glomerular markers (eg. NPHS1, LRP2, UMOD, AQP2). In contrast, we identified a set of markers specific to the interstitium in reference nephrectomy samples. While some of them were novel (eg. FABP2, ADGRD1), others were commonly expressed across all three platforms (LTBP1, ELN, SYNM, ADCY5, COL4A1, C1R). The expression of these genes was localized to expected cell types such as stromal, vascular and immune cells. The renal interstitium from diabetic biopsies revealed differential expression in many pathways including extracellular matrix organization (p=0.0037) and chemokine signaling (p=0.0065).

Conclusion

We successfully isolated the interstitium of human kidney samples using LMD. Gene expression from our samples correlated well with vascular, immune and stromal cell clusters from scRNAseq and snRNAseq data. Therefore, our LMD approach allows rich deconvolution of transcriptomic signatures from single cell datasets and facilitates backmapping of unidentified clusters to the interstitium. Dramatic changes in the gene expression in diabetic kidney interstitium suggest that this compartment may be an important player in the pathophysiology of diabetic nephropathy.

Funding

  • NIDDK Support