Establishment Of Human Heart Biobank And Exploration Of Key Genes Of Coronary Heart Disease And Heart Failure

Heart failure is a global health problem with high morbidity and mortality,which imposes an extremely heavy economic burden on society and families.Heart failure can be caused by a variety of diseases,of which the two most important diseases are coronary atherosclerotic heart disease(ischemic cardiomyopathy)and dilated cardiomyopathy.The molecular mechanism involved in the progression of heart failure is complex and is one of the research hotspots at present.Human biological samples are important carriers to study the molecular mechanism of human diseases,but as life support organs,their tissues are extremely difficult to obtain.At present,the vast majority of biological samples of human heart tissue are obtained from heart transplantation operations.However,due to the complexity of heart surgery,the high cost of surgery and the scarcity of donor hearts,there are still very few heart transplantation operations performed in the world.Since the heart transplant operation was carried out in Guangdong Provincial People's Hospital in 2017,we have established the heart transplant biobank at the same time,and tried to explore the key genes of diseases such as coronary atherosclerotic cardiomyopathy(ischemic cardiomyopathy)and heart failure through these rare samples.To date,we have collected and preserved 172 transplanted heart specimens(including ischemic cardiomyopathy,dilated cardiomyopathy,valvular cardiomyopathy,hypertrophic cardiomyopathy,restrictive cardiomyopathy,cardiac malignant tumor,and post-transplant rejection,etc.)and 5 unused donor heart specimens,which was a huge and valuable cardiovascular resource.Data independent acquisition(DIA)mass spectrometry and RNA-seq technologies were performed in left ventricular species from 5 ICM patients and 5 unused non-failing donors.A total of 546 differentially expressed proteins(DEPs)and 1080 m RNAs(DEGs)were identified in ICM compared with control,which were mainly involved in inflammatory/immune response,response to stress(such as hypoxia and reactive oxidative species),oxidative stress,and ECM(extracellular matrix)organization.Moreover,though the low correlation between transcriptome and proteome,41 key genes were identified,which showed the same expression directions at m RNA and protein levels.Among them,HSP90AA1 occupied a central position in the PPI network.Furthermore,a differentially expressed lnc RNA-m RNA-protein network was constructed,which consisted of 13,11,and11 differentially expressed lnc RNAs,m RNAs,and proteins,respectively.And the expression of this network was validated by q RT-PCR.In addition,we demonstrated that silencing lnc RNA GPAM-203 could inhibit hypoxia-induced apoptosis of AC16 by regulating transferrin(TF).Moreover,in order to further explore the common key genes of ICM and DCM that are closely related to heart failure,we analyzed the network big data.By analyzing the datasets of GSE1869,GSE5406,GSE57338,GSE79962,GSE116250 and GSE46224,Seventy-four coregulated ICM and 126 coregulated DCM relevant DEGs were identified.Moreover,59 common genes between ICM and DCM relevant DEGs were obtained that were mainly involved in cardiac fibrosis and several signal pathways,such as Wnt signal pathway,PI3K-Akt signal pathway,and HIF-1A signal pathway.Among them,six key genes,namely ASPN,FMOD,DPT,CCND1,LUM,and OGN,were at the hub of the PPI network.In addition,through the analysis of the two largest microarray datasets(GSE5406 and GSE57338)containing human heart tissue ICM and DCM by weighted gene co-expression network analysis(WGCNA),we identified one gene module that were highly correlated to heart failure,and identified five heart failure hub genes,namely ASPN,FMOD,NT5 E,LUM and OGN.From two perspectives,we identified four genes that are highly related to heart failure,namely ASPN,FMOD,LUM and OGN.Among the four hub genes,ASPN had a relatively higher correlation with left ventricular ejection fraction(LVEF).Finally,TMA and Western-Blotting results revealed that the ASPN protein was significantly increased in ICM and DCM left ventricular samples.In addition,since atherosclerosis is the cause of ischemic cardiomyopathy and foam cells,which are mainly from smooth muscle cells(SMCs)and macrophages,play a crucial role in all stages of atherosclerosis,we further explored the common molecular mechanisms of these two types of foam cells.By analyzing GSE28829,GSE43292,GSE68021 and GSE54666,we identified a total of 432 SMCs-derived foam cell-related genes and 81macrophage-derived foam cell-related genes,which were mainly involved in lipid metabolism,inflammation,cell cycle/apoptosis.Furthermore,three co-regulated genes associated with foam cells were identified: GLRX,RNF13,and ABCA1.These three common genes showed an increased tendency in unstable or ruptured plaques,although in some cases,no statistically significant difference was found.In addition,immunofluorescence results showed that RNF13 protein expression was increased in atherosclerotic plaques relative to both normal coronary arteries and diseased vascular mediums,and it was co-localized with biomarkers of smooth muscle cells and macrophages.In conclusion,this study explored the key genes of coronary atherosclerotic heart disease and heart failure by using human heart tissue,which has certain guiding significance for future research on prevention and treatment of heart failure.