Weighted Gene Co-Expression Network Analysis Identifies ANGPTL4 as a Key Regulator in Diabetic Cardiomyopathy via FAK/SIRT3/ROS Pathway in Cardiomyocyte.

Lei Dai,Yang Xie,Hongcheng Jiang,Xiaodan Zhong,Wenjun Zhang,Hesong Zeng,Mengwen Wang,Zhen He,Xiaolei Liu,Hongjie Wang

doi:10.3389/fendo.2021.705154

Lei Dai, Yang Xie + Show 8 more

Open Access

https://doi.org/10.3389/fendo.2021.705154

Copy DOI

Abstract

BackgroundDiabetic cardiomyopathy (DbCM) is characterized by initial impairment of left ventricular relaxation followed by contractile dysfunction. Despite intensive research, the exact mechanism remains so far unsolved.MethodsWe constructed weighted gene co-expression network analysis (WGCNA) to screen gene modules that were closely related with DbCM based on the GSE5606 dataset, which contained expression data of the cardiac left ventricle in a rodent model of streptozotocin (STZ)-induced DbCM. Then, the most related hub gene, angiopoietin-like 4 (ANGPTL4), was selected for functional ex vivo and in vitro assays. In our experiments, STZ-induced diabetic mice (C57BL/6J) and human cardiomyocytes (AC16) were used to study the functional roles and potential mechanisms of ANGPTL4 in DbCM.ResultsWGCNA analysis revealed the yellow and green modules were most correlated with DbCM, and identified ANGPTL4 as one of the most significantly upregulated hub genes (ANGPTL4, ACOT1, DECR1, HMGCS2, and PDK4). Consistent with the bioinformatic analysis, the amount of ANGPTL4 was significantly upregulated in diabetic mouse heart. DbCM group, compared with the control group, had increased phosphorylation of focal adhesion kinase (FAK), reduced SIRT3 expression, increased SOD2 acetylation, upregulated NADPH oxidase activation, elevated reactive oxygen species (ROS) produciton, and enhanced apoptosis in the diabetic mouse heart. Moreover, ANGPTL4 induced apoptosis via FAK/SIRT3/ROS pathway in human cardiomyocytes (AC16) under high glucose condition in vitro.These effects were abrogated by treatment of two independent siRNA for ANGPTL4, whereas exogenous recombinant ANGPLT4 protein treatment exacerbated those effects in AC16.ConclusionWe found ANGPTL4, ACOT1, DECR1, HMGCS2, and PDK4 were significantly increased in diabetic heart. ANGPTL4 could promote cardiac apoptosis via a FAK/SIRT3/ROS dependent signaling pathway in DbCM.

Highlights

Diabetes mellitus (DM) is one of the fastest growing global health emergencies of the 21st century
weighted gene co-expression network analysis (WGCNA) was performed to classify the genes with similar expression profiles into the same modules
Module eigengene dendrogram showed that the 14 modules were mainly divided into two clusters, and similar results were demonstrated by the heatmap plotted (Figure 2F)

Summary

Introduction

Diabetes mellitus (DM) is one of the fastest growing global health emergencies of the 21st century. In 2019, it is estimated that 463 million people have diabetes, and this number is projected to reach 578 million by 2030 and 700 million by 2045 [1]. Cardiovascular complications are the primary cause of mortality and morbidity in diabetic patients [2]. Diabetes increases the risk of heart failure approximately two-fold, independent of coronary artery disease and other comorbidities [3]. Rubler et al found that a subset of diabetic patients develops left ventricular dysfunction in the absence of coronary artery disease, hypertension, or vascular disease, and they termed this diabetic cardiomyopathy (DbCM) [4]. Diabetic cardiomyopathy (DbCM) is characterized by initial impairment of left ventricular relaxation followed by contractile dysfunction. The exact mechanism remains so far unsolved

Methods

Results

Conclusion