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Activation of necroptosis pathway in podocyte contributes to the pathogenesis of focal segmental glomerular sclerosis

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Abstract

Background

Focal segmental glomerulosclerosis (FSGS) is characterized by podocyte damage and severe proteinuria. The exact mechanism of podocyte damage and loss remains unclear. Necroptosis, a lytic form of programmed cell death mediated by RIP3 and MLKL, has emerged as an important cell death pattern in multiple tissues and cell types. Necroptosis in FSGS has not been investigated.

Methods

Public GEO data regarding podocyte treated with vehicle or adriamycin (ADR) was identified and analyzed. Cultured human podocytes were used to explore the activation of necroptosis upon ADR stimulation. The expression of necroptosis pathway molecules, p-RIP3 and p-MLKL, was examined in the glomeruli and defoliated urinary podocytes of patients with FSGS. The effect of necroptosis inhibition was assessed in ADR-induced glomerulopathy mice using GSK872.

Results

Publicly available RNA-sequencing data analysis showed that both necroptosis and NLRP3 inflammasome pathway were up-regulated in ADR-injured podocyte. Immunofluorescent staining showed increased expression of p-RIP3 and p-MLKL, the active forms of RIP3 and MLKL, in podocytes of FSGS patients and ADR-induced glomerulopathy mice but not in the normal control. GSK872, an RIP3 kinase inhibitor, significantly inhibited the expression of p-RIP3, p-MLKL and activation of NLRP3 in cultured podocytes treated with ADR. GSK872 treatment of mice with ADR-induced nephropathy resulted in the reduced expression of p-RIP3, p-MLKL, NLRP3 and caspase-1 p20. GSK872 also significantly inhibited the expression of p-MLKL in the podocytes of ADR-induced nephropathy, resulting in the attenuation of proteinuria and renal histological lesions.

Conclusion

Necroptosis pathway might be a valuable target for the treatment of FSGS.

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Funding

This study was supported by the Dongguan Science and Technology of Social Development Program (201950715001183).

Author information

Author notes

  1. Haoqiang Hu and Mengyuan Li have contributed equally to this work.

Authors and Affiliations

  1. Department of Nephrology, Affiliated Dongguan People’s Hospital, Southern Medical University (Dongguan People’s Hospital), Dongguan, 523000, China

    Haoqiang Hu

  2. Department of Rheumatology, The First Affiliated Hospital, Sun Yat-Sen University, 58 Zhongshan Second Road, Guangzhou, 510080, China

    Haoqiang Hu, Mengyuan Li, Binfeng Chen, Chaohuan Guo & Niansheng Yang

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  1. Haoqiang Hu
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  2. Mengyuan Li
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  5. Niansheng Yang
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Contributions

All authors were involved in drafting and revising the article. NY, HH, ML: conceived and designed the study, HH and ML: collected clinical samples, HH, ML, CG, and BC: conducted experiments, NY, ML, CG and BC: analyzed and interpreted data, ML and NY: wrote the manuscript.

Corresponding author

Correspondence to Niansheng Yang.

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Conflict of interest

The authors have declared that no conflict of interest exists.

Ethical approval

This study was approved by the Institutional Research Ethics Committee of Affiliated Dongguan People’s Hospital, Southern Medical University, China (IRB approval number KYKT2021-034). This study was performed in compliance with the Helsinki Declaration. Volunteer patients provided written consent. All procedures involving experimental animals were approved by the Ethical Committee of Animal Research of the Sun Yat-sen University (IRB approval number SYSU-IACUC-2020-000151).

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Hu, H., Li, M., Chen, B. et al. Activation of necroptosis pathway in podocyte contributes to the pathogenesis of focal segmental glomerular sclerosis. Clin Exp Nephrol 26, 1055–1066 (2022). https://doi.org/10.1007/s10157-022-02258-1

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  • DOI: https://doi.org/10.1007/s10157-022-02258-1

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