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The role of PCSK9 in glomerular lipid accumulation and renal injury in diabetic kidney disease

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Abstract

Aims/hypothesis

Glomerular lipid accumulation is a defining feature of diabetic kidney disease (DKD); however, the precise underlying mechanism requires further elucidation. Recent evidence suggests a role for proprotein convertase subtilisin/kexin type 9 (PCSK9) in intracellular lipid homeostasis. Although PCSK9 is present in kidneys, its role within kidney cells and relevance to renal diseases remain largely unexplored. Therefore, we investigated the role of intracellular PCSK9 in regulating lipid accumulation and homeostasis in the glomeruli and podocytes under diabetic conditions. Furthermore, we aimed to identify the pathophysiological mechanisms responsible for the podocyte injury that is associated with intracellular PCSK9-induced lipid accumulation in DKD.

Methods

In this study, glomeruli were isolated from human kidney biopsy tissues, and glomerular gene-expression analysis was performed. Also, db/db and db/m mice were used to perform glomerular gene-expression profiling. We generated DKD models using a high-fat diet and low-dose intraperitoneal streptozocin injection in C57BL/6 and Pcsk9 knockout (KO) mice. We analysed cholesterol and triacylglycerol levels within the kidney cortex. Lipid droplets were evaluated using BODIPY staining. We induced upregulation and downregulation of PCSK9 expression in conditionally immortalised mouse podocytes using lentivirus and siRNA transfection techniques, respectively, under diabetic conditions.

Results

A significant reduction in transcription level of PCSK9 was observed in glomeruli of individuals with DKD. PCSK9 expression was also reduced in podocytes of animals under diabetic conditions. We observed significantly higher lipid accumulation in kidney tissues of Pcsk9 KO DKD mice compared with wild-type (WT) DKD mice. Additionally, Pcsk9 KO mouse models of DKD exhibited a significant reduction in mitochondria number vs WT models, coupled with a significant increase in mitochondrial size. Moreover, albuminuria and podocyte foot process effacement were observed in WT and Pcsk9 KO DKD mice, with KO DKD mice displaying more pronounced manifestations. Immortalised mouse podocytes exposed to diabetic stimuli exhibited heightened intracellular lipid accumulation, mitochondrial injury and apoptosis, which were ameliorated by Pcsk9 overexpression and aggravated by Pcsk9 knockdown in mouse podocytes.

Conclusions/interpretation

The downregulation of PCSK9 in podocytes is associated with lipid accumulation, which leads to mitochondrial dysfunction, cell apoptosis and renal injury. This study sheds new light on the potential involvement of PCSK9 in the pathophysiology of glomerular lipid accumulation and podocyte injury in DKD.

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Abbreviations

ABCA1:

ATP-binding cassette transporter A1

BAX:

B cell lymphoma-2-associated X

Bcl-2:

B cell lymphoma-2

CMV:

Cytomegalovirus

DKD:

Diabetic kidney disease

GBM:

Glomerular basement membrane

HBSS:

Hanks’ Balanced Salt Solution

HFD:

High-fat diet

IgAN:

IgA nephropathy

IRB:

Institutional Review Board

KO:

Knockout

LDL-C:

LDL-cholesterol

LDLR:

LDL-cholesterol receptor

Lv-PCSK9:

CMV-mPCSK9 lentivirus

PCSK9:

Proprotein convertase subtilisin/kexin type 9

PGC-1α:

Peroxisome proliferator-activated receptor, gamma, coactivator 1, alpha

qRT-PCR:

Quantitative real-time PCR

siPcsk9 :

Pcsk9 siRNA

STZ:

Streptozocin

TEM:

Transmission electron microscopy

TU:

Transfection units

WT:

Wild-type

ZO-1:

Zonula occludens-1

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Authors and Affiliations

  1. Department of Nephrology, The First Hospital of Jilin University, Changchun, China

    Meiyan Wu

  2. Department of Internal Medicine, College of Medicine, Institute of Kidney Disease Research, Yonsei University, Seoul, South Korea

    Meiyan Wu, Chang-Yun Yoon, Jimin Park, Gyuri Kim, Bo Young Nam, Seonghun Kim, Jung Tak Park, Seung Hyeok Han, Shin-Wook Kang & Tae-Hyun Yoo

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  1. Meiyan Wu
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Correspondence to Tae-Hyun Yoo.

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Acknowledgements

We especially thank S. W. Park (Department of Biochemistry and Molecular Biology, Yonsei University College of Medicine, Seoul, South Korea) for providing experimental mice. Some of the results presented in the current article were included in the PhD thesis by Jimin Park. The graphical abstract was created using scidraw.io.

Data availability

The data supporting the findings of the present study are available from the corresponding author upon reasonable request.

Funding

This work was supported by grants from the Korea Healthcare Technology R&D Project, Ministry of Health and Welfare, Republic of Korea (H14C2003000016), the National Research Foundation of Korea (NRF-2019R1A6A3A13094020), the National Natural Science Foundation of China (82000689), the Postdoctoral Science Foundation of China (2021M691212) and the Natural Science Foundation of Jilin Province, China (20210101455JC).

Authors’ relationships and activities

The authors declare that there are no relationships or activities that might bias, or be perceived to bias, their work.

Contribution statement

T-HY conceived and supervised the project and designed the experiments. MW and C-YY performed all experiments. JP, GK and BYN performed sample preparation for TEM, image acquisition and data analysis. SK conducted BODIPY 493/503 staining and subsequent image analysis. MW and C-YY interpreted data. T-HY, C-YY and MW wrote the manuscript. JTP, SHH and S-WK reviewed the article critically for important intellectual content. All authors contributed to the analysis and discussion of the data. All authors revised and approved the manuscript in its final form. T-HY is the guarantor of this work.

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Wu, M., Yoon, CY., Park, J. et al. The role of PCSK9 in glomerular lipid accumulation and renal injury in diabetic kidney disease. Diabetologia (2024). https://doi.org/10.1007/s00125-024-06191-8

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