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Abstract: SA-PO574

Cell Fraction Changes Show Strong Correlation with Kidney Disease Severity in Mice and in Patients

Session Information

Category: Glomerular Diseases

  • 1201 Glomerular Diseases: Fibrosis and Extracellular Matrix

Authors

  • Yang, Yawen, University of Pennsylvania, Philadelphia, Pennsylvania, United States
  • Park, Jihwan, University of Pennsylvania, Philadelphia, Pennsylvania, United States
  • Shrestha, Rojesh, University of Pennsylvania, Philadelphia, Pennsylvania, United States
  • Han, Seungyub, University of Pennsylvania, Philadelphia, Pennsylvania, United States
  • Sheng, Xin, University of Pennsylvania, Philadelphia, Pennsylvania, United States
  • Li, Mingyao, University of Pennsylvania, Philadelpiha, Pennsylvania, United States
  • Kim, Junhyong, University of Pennsylvania, Philadelphia, Pennsylvania, United States
  • Susztak, Katalin, University of Pennsylvania, Philadelphia, Pennsylvania, United States

Group or Team Name

  • SUSZTAK LAB
Background

A revolution in cellular measurement technology is under way. While prior studies have only been able to analyze averaged outputs from whole kidneys, now we can accurately monitor genome-wide gene expression, regulation, function, cellular history and cellular interactions in thousands of individual cells in a single experiment. We used single cell RNA analysis to unravel the kidney and immune cell diversity and cell plasticity in healthy and diseased kidneys.

Methods

Five to 8-week-old male FvB wild-type mice underwent ligation of the left ureter and were sacrificed on day 7. Single cell RNA sequencing was performed using the 10X Genomics Chromium System. Data was aligned using Cell Ranger Software and processed using the Seurat package.

Results

We analyzed the transcriptome of 59,609 individual cells from 6 control and 2 UUO kidneys. We identified 30 different major cell populations and multiple subpopulations. Specifically, we found that immune cell diversity was much greater in disease tissue compared to control animals and we identified 16 different immune cell types, including neutrophils, basophils, macrophages, B-cells, plasma cells, CD4 T cells, Th17 cells, T regulatory cells, NK cells, CD8 cells, CD8 effector cells, dendritic cells and plasmacytoid dendritic cells. We found that the fraction of immune cells were also significantly increased in UUO kidneys by single cell analysis. In silico cell deconvolution analysis of bulk RNA sequencing further confirmed loss of epithelial cells and increased in immune cell numbers in UUO mice. Cell-cell interaction analysis indicated that cytokines secreted by injured epithelial cells play key role in immune cell recruitment into disease samples, for example the expression of CD34 responsible for myeloid cell recruitment, Cxc10 and Cxc16 for lymphocyte recruitment. In silico cell deconvolution analysis of 91 human kidney biopsy samples indicated significant cell proportion changes in patient samples. We found that cell proportion changes correlated with the degree of kidney disease such as GFR or fibrosis.

Conclusion

Comparing to health kidneys, UUO kidneys are characterized by increase cell diversity and cell type specific changes. Cell fraction changes show strong correlation with the degree of fibrosis in mice and in patients.

Funding

  • NIDDK Support