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Abstract: SA-PO1021

The Expression Profile of Complements in Human Renal Mesangial Cells, Endothelial Cells, Podocytes, and Proximal Tubular Epithelial Cells

Session Information

Category: Glomerular Diseases

  • 1403 Podocyte Biology

Authors

  • Gao, Yueming, Department of Nephrology, Peking University Third Hospital, Beijiing, China
  • Ma, Jin, Department of Nephrology, Peking University Third Hospital, Beijiing, China
  • Deng, Zhenling, Department of Nephrology, Peking University Third Hospital, Beijiing, China
  • Wang, Yue, Department of Nephrology, Peking University Third Hospital, Beijiing, China
Background

More and more evidence indicated that the kidney was a notable source of complements. We aimed to explore the expression profile of complements in human renal mesangial cell (HRMC), glomerular endothelial cell (HRGEC), podocyte (HPC), and proximal tubular epithelial cell (PTEC) under physiological conditions.

Methods

Single-cell RNA sequencing (scRNA-seq) by a 10xGenomics Chromium system was used to identify the transcription of 30 complement genes in HRMCs, HRGECs, HPCs, and PTECs. RT-PCR was used to verify the expression of complement genes in cultured primary HRMC and HRGEC, and in HPC and PTEC cell lines. Immunofluorescence was used to confirm the protein expression of several key complements.

Results

By scRNA-seq, the transcription of nearly all complement genes was found in HRMCs, HRGECs, HPCs, and PTECs, and the expression of complement regulatory proteins was highest. RT-PCR confirmed the transcription of C1S, C1R, C4, CFD, MBL2, MASP1, MASP2, C3, C5, CFI, DAF, CD46, CD59, protein S, and C3AR1 in cultured cells. Immunofluorescence verified the expression of C3, DAF, CD46, C4BPB, and CD59.

Conclusion

Under physiological conditions, HRMCs, HRGECs, HPCs, and PTECs express multiple complements involved in three activation pathways.

Figure 1. The expression profile of complement genes identified by scRNA-seq and RT-PCR

Figure 2. Complement protein expression in cultured human renal cells

Funding

  • Government Support – Non-U.S.