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YAP mediates the interaction between the Hippo and PI3K/Akt pathways in mesangial cell proliferation in diabetic nephropathy

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Abstract

Aims

Glomerular mesangial cell (MC) proliferation is one of the main pathological changes in diabetic nephropathy (DN), but its mechanism needs further elaboration. The Hippo and PI3K/Akt signalling pathways are involved in the regulation of MC proliferation, but their relationship in hyperglycaemia-induced MC proliferation has not been reported.

Methods

We used db/db mice and high-glucose-cultured mesangial cells to generate a diabetic nephropathy model. An MST1-knockdown plasmid was used to identify whether the PI3K/Akt pathway is linked to the Hippo pathway through MST1. LY294002 and SC79 were used to verify the role of the PI3K/Akt signalling pathway in MC cells. RNA silencing and overexpression were performed by using YAP and PTEN-expression/knockdown plasmids to investigate the function of YAP and PTEN, respectively, in the Hippo and PI3K/Akt signalling pathways.

Results

By examining a potential feedback loop, we found decreased phosphorylation of MST1 and Lats1 and increased PI3K/Akt activation in db/db mice and high glucose-treated MCs, along with increased MC proliferation. The results of our gene silencing experiment proved PI3K/Akt-mediated intervention in the Hippo pathway and the regulatory effect of YAP on PI3K/Akt through PTEN.

Conclusions

The Hippo pathway is inhibited under diabetic conditions, leading to YAP activation and promoting MC proliferation. The PI3K/Akt pathway is activated through the inhibitory effect of YAP on its repressor, PTEN. Finally, activation of the PI3K/Akt pathway inhibits the Hippo pathway, resulting in nuclear YAP accumulation and accelerating MC proliferation and DN formation.

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Abbreviations

BUN:

Blood urea nitrogen

Cr:

Creatinine

DN:

Diabetic nephropathy

ECM:

Extracellular matrix

FBG:

Fasting blood glucose

Lats1/2:

Large tumour suppressor 1/2 serine/threonine protein kinases

MC:

Mesangial cell

MOB1:

Mps-one binder 1

MST1/2:

Module consists of Ste20-like serine/threonine kinases 1/2

PAS:

Periodic acid Schiff

PI3K:

Phosphoinositide-3-kinase

TAZ:

Transcriptional co-activator with PDZ binding motif

TBST:

Tris-buffered saline with Tween

TEAD:

TEA domain

YAP:

Yes-associated protein

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Acknowledgements

The work was supported by the National Natural Science Foundation of China (Grant No. 81473257, 81400741), the Qing Lan project, the Natural Science Foundation of Jiangsu Province (Grant No. BK20151155), the “333” Foundation of Jiangsu Province (Grant No. BRA2015329), the Key Natural Science Foundation of Jiangsu Higher Education Institutions of China (Grant No.15KJA310005), Jiangsu Overseas Research & Training Program for University Young & Middle-aged Teachers and Presidents, Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD), and Postgraduate Research and Innovation Program of Xuzhou Medical University of Pharmacy (Grant No. KYCX17_1711).

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Author notes

  1. Xuan Qian, Linlin He and Meng Hao have contributed equally to this study.

Authors and Affiliations

  1. Jiangsu Key Laboratory of New Drug Research and Clinical Pharmacy, Xuzhou Medical University, Xuzhou, Jiangsu, China

    Xuan Qian, Linlin He, Meng Hao, Yuan Li, Xizhi Li, Yiqi Liu, Hong Jiang, Liu Xu, Chengcheng Li, Wenya Wu, Lei Du, Xiaoxing Yin & Qian Lu

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  1. Xuan Qian
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Contributions

LD, QL, and XY participated in research design. XQ, YL, CL, XL, HJ, YL, LX, and WW conducted experiments. XQ, LH, HJ, YL, LX, and WW performed data analysis. XQ, LH, MH, YL, LD, QL, and XY wrote or contributed to the writing of the manuscript.

Corresponding author

Correspondence to Qian Lu.

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Conflict of interest

The authors have no confict of interest to declare.

Ethical approval

Animal experiments were conducted in accordance with the principles provided by the National Institutes of Health (NIH) Guideline for the Care and Use of Laboratory Animals. Approval to proceed with this experiment was issued by the Animal Ethics Committee (201707w012) of Xuzhou Medical University, and protocols also conformed to the Guidelines for Ethical Conduct in the Care and Use of Animals.

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For this type of study informed consent is not required.

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This article belongs to the topical collection Diabetic Nephropathy, managed by Giuseppe Pugliese.

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Fig. S1

The plasmids are efficient, and the siControl plasmids have no significant effect on the expression of relative proteins. (a) Expression of YAP in mouse MCs by Western blotting and statistical result of YAP protein expression. (b) Expression of Mst1 in mouse MCs by Western blotting and statistical result of Mst1 protein expression. (c) Expression and statistical results of relative proteins. Data are expressed as mean ± SD, n = 3. (PDF 156 kb)

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Qian, X., He, L., Hao, M. et al. YAP mediates the interaction between the Hippo and PI3K/Akt pathways in mesangial cell proliferation in diabetic nephropathy. Acta Diabetol 58, 47–62 (2021). https://doi.org/10.1007/s00592-020-01582-w

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