Kidney Week

Abstract: TH-PO416

Single-Cell Transcriptomics Indicate That CD163+ Kidney Macrophages Are Pro-Inflammatory and May Orchestrate Further Immune Cell Recruitment to Kidney Lesions in ANCA-Associated Glomerulonephritis

Session Information

• Glomerular Diseases: Inflammation and Fibrosis
  November 03, 2022 | Location: Exhibit Hall, Orange County Convention Center‚ West Building
  Abstract Time: 10:00 AM - 12:00 PM

Category: Glomerular Diseases

• 1302 Glomerular Diseases: Immunology and Inflammation

Authors

• Vegting, Yosta, Amsterdam UMC location University of Amsterdam, Department of Internal Medicine, Section of Nephrology, Amsterdam, Netherlands

Yosta Vegting,

• Jongejan, Aldo, Amsterdam UMC location University of Amsterdam, Department of Epidemiology & Data Science (EDS), Bioinformatics Laboratory, Amsterdam, Netherlands

Aldo Jongejan,

• Remmerswaal, Ester B.M., Amsterdam UMC location University of Amsterdam, Department of Internal Medicine, Section of Nephrology, Amsterdam, Netherlands

Ester B.M. Remmerswaal,

• Moerland, Perry, Amsterdam UMC location University of Amsterdam, Department of Epidemiology & Data Science (EDS), Bioinformatics Laboratory, Amsterdam, Netherlands

Perry Moerland,

• Kers, Jesper, Amsterdam UMC location University of Amsterdam, Department of Pathology, Amsterdam, Netherlands

Jesper Kers,

• Vogt, Liffert, Amsterdam UMC location University of Amsterdam, Department of Internal Medicine, Section of Nephrology, Amsterdam, Netherlands

Liffert Vogt,

• Bemelman, Frederike J., Amsterdam UMC location University of Amsterdam, Department of Internal Medicine, Section of Nephrology, Amsterdam, Netherlands

Frederike J. Bemelman,

• Hilhorst, Marc, Amsterdam UMC location University of Amsterdam, Department of Internal Medicine, Section of Nephrology, Amsterdam, Netherlands

Marc Hilhorst,

Background

Anti-Neutrophil Cytoplasmic Antibodies (ANCA)-associated glomerulonephritis (AGN) affects up to 90% of patients with ANCA-vasculitis and may result in end-stage kidney disease. In AGN kidney biopsies, CD163⁺ macrophages are abundantly present and linked to disease activity. However, their exact role in the maintenance or resolution of local inflammation remains unclear.

Methods

CD163⁺ kidney macrophages were studied using single-cell RNA sequencing of fresh kidney biopsies of 3 active ANCA patients and one patient receiving a nephrectomy for renal cell carcinoma. Cells were sorted for living (CD45⁺) immune cells by flowcytometry and whole transcriptome analysis was performed using 10X Genomics’ technology. To aid in the multiomic characterization, 2 samples were incubated with an oligo-antibody cocktail and T and B Cell Receptor Repertoires were sequenced.

Results

Over 29,000 cells were sequenced: 2,300 (8%) were identified as CD68⁺ macrophages. In patient and control samples, 498/19,689 (2.5%) and 320/9,605 (3.3%) cells expressed CD163, respectively. Comparison of gene-expression between CD163⁺ macrophages of the two groups showed significant upregulation of pro-inflammatory cytokines and chemokines (Log2FoldChange ANCA vs control) (IL1B 1.62, CXCL2 2.25, CXCL3 3.26, CXCL8 2.19, CCL3 1.81, CCL4 2.04), and the complement system (C1Qa 1.79, C1Qb 1.94, C1Qc 1.37, C5aR1 1.43). Expression of CCL17 was downregulated (CCL17 -6.57).

Conclusion

CD163⁺ kidney macrophages are pro-infammatory and may orchestrate further immune cell recruitment to active kidney lesions in AGN.

Funding

• Private Foundation Support