Kidney Week

Abstract: FR-OR31

The Single-Cell Landscape of Kidney Immune Cells Reveals Transcriptional Heterogeneity in Late Diabetic Kidney Disease

Session Information

• Diabetic Kidney Disease: From Single Cell to Outcomes
  November 04, 2022 | Location: W240, Orange County Convention Center‚ West Building
  Abstract Time: 04:30 PM - 04:39 PM

Category: Diabetic Kidney Disease

• 601 Diabetic Kidney Disease: Basic

Authors

• Fu, Jia, Icahn School of Medicine at Mount Sinai, New York, New York, United States

Jia Fu, MD, MS

• Sun, Zeguo, Icahn School of Medicine at Mount Sinai, New York, New York, United States

Zeguo Sun,

• Zhang, Weijia, Icahn School of Medicine at Mount Sinai, New York, New York, United States

Weijia Zhang,

• Lee, Kyung, Icahn School of Medicine at Mount Sinai, New York, New York, United States

Kyung Lee,

• He, John Cijiang, Icahn School of Medicine at Mount Sinai, New York, New York, United States

John Cijiang He,

Background

Mounting evidence supports the involvement of renal inflammation is the key driver in diabetic kidney disease (DKD) pathogenesis and it contributes significantly to the progression of DKD. Many studies have demonstrated that macrophages are a major inflammatory cell type infiltrating the kidney in both patient and experimental animal models of DKD. Our preliminary data revealed relatively mild alteration in molecular and phenotype shift in macrophage subpopulation in early DKD, but the dynamic shift and transcriptional signature of these subpopulations in later stages of DKD remain largely unknown.

Methods

Fluorescence-activated cell sorting was used to isolate CD45+ cells from control and OVE26 diabetic mice at 28 weeks. To uncover the gene expression changes in specific immune cell subsets in late DKD, we employed single-cell RNA sequencing using the 10x Genomics platform. We generated single cell transcriptome profiles and performed comprehensive bioinformatics analysis and gene expression validation. CellRanger, R and Seurat, Macspectrum were used to make gene-cell matrices and for downstream analyses.

Results

Gene ontology enrichment analysis showed pronounced activation of immune pathways shared across MNPs in late DKD, whereas oxidative phosphorylation and cellular respiration were enriched in early DKD. Compared to early DKD, there’s a significant accumulation of infiltrating macrophage (IM) in late DKD, particularly anti-inflammatory Ly6c^low Ace^hi IMs and a novel population M2-like macrophage, which highly expressed alternatively activated macrophage markers (Retnla, Fcna,Cd163). After injury, go terms show Ly6c^low IM played more roles in tissue repair, regulation of cell matrix adhesion, and cellular response to laminar fluid shear stress upon injury. Interestingly, M2-like macrophages were well distinguished by expressing genes enriched in TGF-β signaling, a key pathway that leads to renal fibrosis.

Conclusion

We revealed recruitment of alternatively activated macrophage and reparative macrophage phenotype Ly6c^low Ace^hi IMs in late DKD, suggesting the dynamic plasticity of macrophage functions in kidney during disease progression. This highlights the value of potential therapeutic target of immunotherapy as a means of preventing the progression of DKD are discussed.

Funding

• NIDDK Support