Genetic variants of VEGFR-1 gene promoter in acute myocardial infarction

Abstract Background Coronary artery disease (CAD) including acute myocardial infarction (AMI) is a common complex disease caused by atherosclerosis. Vascular epithelial growth factor receptor-1 (VEGFR-1) stimulates angiogenesis and vascular permeability, and functions as a decoy to sequester VEGF and prevent initiation of intracellular signaling. VEGFR-1 knockout mice exhibit significantly higher mortality due to heart failure, cardiac hypertrophy, and cardiac dysfunction. An evident increase in macrophage infiltration and cardiac fibrosis are also observed after transverse aortic constriction. Therefore, VEGFR-1 gene variants may be involved in CAD. In this study, VEGFR-1 gene promoter was genetically and functionally analyzed in large cohorts of AMI patients and ethnic-matched controls. Results A total of 16 DNA sequence variants (DSVs) including six single-nucleotide polymorphisms (SNPs) were found in the VEGFR-1 gene promoter and 5′-untranslated region. Five novel DSVs and one SNP were only identified in AMI patients group. These DSVs and SNP significantly altered the transcriptional activity of the VEGFR-1 gene promoter in both HEK-293 and H9c2 cells (P < 0.05). Further electrophoretic mobility shift assay indicated that the DSVs and SNPs evidently affected the binding of transcription factors. Conclusions The genetic variants in VEGFR-1 gene identified in AMI patients may alter the transcriptional activity of the VEGFR-1 gene promoter and change VEGFR-1 level, contributing to AMI development.

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Functional Genetic Variant in ATG5 Gene Promoter in Acute Myocardial Infarction

Cardiology Research and Practice ◽

10.1155/2020/9898301 ◽

2020 ◽

Vol 2020 ◽

pp. 1-7

Author(s):

Yexin Zhang ◽

Xiaohui He ◽

Jiarui Li ◽

Wentao Yang ◽

Yinghua Cui ◽

...

Keyword(s):

Myocardial Infarction ◽

Acute Myocardial Infarction ◽

Genetic Variants ◽

Transcriptional Activity ◽

Genetic Variant ◽

Core Protein ◽

Gene Promoter ◽

Mobility Shift ◽

Hek 293 ◽

Deletion Variant

Coronary artery disease (CAD) including acute myocardial infarction (AMI) is an inflammatory and metabolic disease mainly caused by atherosclerosis. Dysfunctional autophagy has been associated with abnormal lipid metabolism and inflammation. In previous studies, we have reported altered autophagic activity in AMI patients. As autophagy-related protein 5 (ATG5) is a core protein in autophagy, we speculated that altered ATG5 level may contribute to CAD and AMI development. In this study, the promoter of the ATG5 gene was genetically and functionally investigated in large groups of AMI patients (n = 378) and ethnic-matched healthy controls (n = 386). The results showed that a total of 15 genetic variants including 6 single-nucleotide polymorphisms (SNPs) in the ATG5 gene promoter were found in this study population. A novel deletion variant (g.106326168_70delTCT) and an SNP [g.106325757C > G (rs190825454)] were found in one 66-year-old male patient with non-ST-segment elevated AMI, but in none of controls. In cultured HEK-293 and H9c2 cells, the deletion variant significantly decreased the transcriptional activity of the ATG5 gene promoter (P<0.01). In contrast, the genetic variants either identified only in controls or found in both AMI patients and controls did not affect the transcriptional activity of the ATG5 gene promoter (P>0.05). Furthermore, an electrophoretic mobility shift assay showed that the deletion variant evidently affected the binding of a transcription factor. Therefore, the genetic variant identified in AMI may affect the activity of the ATG5 gene promoter and change the ATG5 level, contributing to AMI as a rare risk factor.

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Identification and functional analysis of genetic variants in TBX5 gene promoter in patients with acute myocardial infarction

BMC Cardiovascular Disorders ◽

10.1186/s12872-019-1237-6 ◽

2019 ◽

Vol 19 (1) ◽

Cited By ~ 1

Author(s):

Shuai Wang ◽

Jie Zhang ◽

Xiaohui He ◽

Yexin Zhang ◽

Jing Chen ◽

...

Keyword(s):

Myocardial Infarction ◽

Acute Myocardial Infarction ◽

Transcription Factors ◽

Complex Disease ◽

Cultured Cells ◽

Normal Population ◽

Gene Promoter ◽

Epidemiological Studies ◽

Common Complex Disease ◽

Tbx5 Gene

Abstract Background Coronary artery disease (CAD), including acute myocardial infarction (AMI), is a common complex disease. Although a great number of genetic loci and variants for CAD have been identified, genetic causes and underlying mechanisms remain largely unclear. Epidemiological studies have revealed that CAD incidence is strikingly higher in patients with congenital heart disease than that in normal population. T-box transcription factors play critical roles in embryonic development. In particular, TBX5 as a dosage-sensitive regulator is required for cardiac development and function. Thus, dysregulated TBX5 gene expression may be involved in CAD development. Methods TBX5 gene promoter was genetically and functionally analysed in large groups of AMI patients (n = 432) and ethnic-matched healthy controls (n = 448). Results Six novel heterozygous DNA sequence variants (DSVs) in the TBX5 gene promoter (g.4100A > G, g.4194G > A, g.4260 T > C, g.4367C > A, g.4581A > G and g.5004G > T) were found in AMI patients, but in none of controls. These DSVs significantly changed the activity of TBX5 gene promoter in cultured cells (P < 0.05). Furthermore, three of the DSVs (g.4100A > G, g.4260 T > C and g.4581A > G) evidently modified the binding sites of unknown transcription factors. Conclusions The DSVs identified in AMI patients may alter TBX5 gene promoter activity and change TBX5 level, contributing to AMI development as a rare risk factor.

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Genetic Variants and Functional Analyses of the ATG16L1 Gene Promoter in Acute Myocardial Infarction

Frontiers in Genetics ◽

10.3389/fgene.2021.591954 ◽

2021 ◽

Vol 12 ◽

Author(s):

Falan Han ◽

Shuchao Pang ◽

Zhaoqing Sun ◽

Yinghua Cui ◽

Bo Yan

Keyword(s):

Myocardial Infarction ◽

Acute Myocardial Infarction ◽

Transcription Factors ◽

Electrophoretic Mobility ◽

Transcriptional Activity ◽

Gene Promoter ◽

Luciferase Reporter ◽

Mobility Shift ◽

Genotype Frequencies ◽

Promoter Mutations

BackgroundAcute myocardial infarction (AMI), a common complex disease caused by an interaction between genetic and environmental factors, is a serious type of coronary artery disease and is also a leading cause of death worldwide. Autophagy-related 16-like 1 (ATG16L1) is a key regulatory factor of autophagy and plays an important role in induced autophagy. In the cardiovascular system, autophagy is essential to preserve the homeostasis and function of the heart and blood vessels. No studies have hitherto examined the association between AMI and ATG16L1 gene promoter.MethodsWe conducted a case-control study, using polymerase chain reaction and sequencing techniques, dual luciferase reporter assay, and electrophoretic mobility shift assay, to analyze genetic and functional variation in the ATG16L1 gene promoter between AMI and controls. A variety of statistical analyses were used to analyze the allele and genotype frequencies and the relationship between single-nucleotide polymorphisms (SNPs) and AMI.ResultsIn all, 10 SNPs and two DNA-sequence variants (DSVs) were identified in 688 subjects, and three ATG16L1 gene promoter mutations [g.233250693 T > C (rs185213911), g.233250946 G > A (rs568956599), g.233251133 C > G (rs1301744254)] that were identified in AMI patients significantly altered the transcriptional activity of ATG16L1 gene promoter in HEH2, HEK-293, and H9c2 cells (P < 0.05). Further electrophoretic mobility shift assays indicated that the SNPs affected the binding of transcription factors (P < 0.01).ConclusionATG16L1 gene promoter mutations in AMI patients may affect the binding of transcription factors and change the transcriptional activity of the ATG16L1 gene, changing the level of autophagy and contributing to the occurrence and development of AMI as rare and low-frequency risk factors.

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Identification of TFEB gene genetic variants in acute myocardial infarction

10.21203/rs.3.rs-23023/v1 ◽

2020 ◽

Author(s):

Jie Zhang ◽

Yexin Zhang ◽

Xiaohui He ◽

Shuai Wang ◽

Shuchao Pang ◽

...

Keyword(s):

Myocardial Infarction ◽

Acute Myocardial Infarction ◽

Lipid Metabolism ◽

Genetic Variants ◽

Cultured Cells ◽

Nucleotide Polymorphisms ◽

Lysosomal Hydrolases ◽

Mobility Shift ◽

Altered Level ◽

Lysosomal System

Abstract Background: Abnormal lipid metabolism and inflammation play critical roles in the initiation and progression of atherosclerosis and its associated complications, including coronary artery disease (CAD) and acute myocardial infarction (AMI). Autophagic-lysosomal system is involved in many physiological processes, such as lipid metabolism and inflammation. TFEB, a master regulator of the system, coordinates the expression of lysosomal hydrolases, lysosomal membrane proteins, and autophagic proteins. Altered level of TFEB gene expression and subsequent changes of autophagic-lysosomal system may be involved in the onset of CAD and AMI.Methods: In this study, the promoter of the TFEB gene was genetically and functionally analyzed in AMI patients (n=352) and ethnic-matched healthy controls (n=337).Results: A total of fifteen genetic variants, including eight single nucleotide polymorphisms (SNPs), were identified in the participants. Two novel genetic variants and four SNPs were only identified in six AMI patients, and significantly altered the transcriptional activity of the TFEB gene in cultured cells. Further electrophoretic mobility shift assay revealed that two genetic variants (g.41737144A>G and g.41736544C>T) and two SNPs [g.41737274T>C (rs533895008) and g.41736987C>T (rs760293138)] evidently affected the binding of transcription factors.Conclusions: Our findings suggested that the genetic variants in TFEB gene promoter may change TFEB levels, contributing to AMI as a low-frequency risk factor.

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Functional genetic variants in the SIRT5 gene promoter in acute myocardial infarction

Gene ◽

10.1016/j.gene.2018.07.010 ◽

2018 ◽

Vol 675 ◽

pp. 233-239 ◽

Cited By ~ 3

Author(s):

Lu Chen ◽

Haiyan Wang ◽

Feng Gao ◽

Jie Zhang ◽

Yexin Zhang ◽

...

Keyword(s):

Myocardial Infarction ◽

Acute Myocardial Infarction ◽

Genetic Variants ◽

Gene Promoter

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Molecular genetic study on GATA5 gene promoter in acute myocardial infarction

PLoS ONE ◽

10.1371/journal.pone.0248203 ◽

2021 ◽

Vol 16 (3) ◽

pp. e0248203

Author(s):

Zhipeng Song ◽

Lu Chen ◽

Shuchao Pang ◽

Bo Yan

Keyword(s):

Myocardial Infarction ◽

Acute Myocardial Infarction ◽

Transcriptional Activity ◽

Molecular Genetic ◽

Farnesoid X Receptor ◽

Neonatal Rat ◽

Molecular Genetic Study ◽

Mobility Shift ◽

Rat Cardiomyocytes ◽

Hek 293

Background Acute myocardial infarction (AMI) is a severe type of coronary artery disease, caused by coronary occlusion and followed by cardiac ischaemia. GATA binding protein 5 (GATA5) is an important member of GATA family and plays an important role in vascular inflammation, endothelial function, oxidative stress and cell metabolism. Previous studies have shown that the DNA sequence variants (DSVs) in GATA4 and GATA6 promoter can increase susceptibility to AMI. In this study, we explored the relationship between GATA5 promoter and AMI for the first time, hoping to provide a new genetic basis for understanding the pathogenesis of AMI. Methods GATA5 promoter was sequenced in 683 individuals (332 AMI patients and 351 controls). The transcriptional activity of the GATA5 promoter with or without DSVs in HEK-293 cells, H9c2 cells and primary neonatal rat cardiomyocytes were examined by Promega Dual-Luciferase® Reporter Assay system. Electrophoretic mobility shift assay (EMSA) was performed to explore whether the DSVs interfered with the binding of transcription factors (TFs). Results Nine mutations have been found in GATA5 promoter, eight of them evidently altered the transcriptional activity of the GATA5 promoter, five of them disrupted the binding of TFs (such as farnesoid X receptor). Furthermore, haplotype AT (across rs80197101 and rs77067995) is a dangerous haplotype of AMI. Genotype GA and allele A of rs80197101 and genotype CT and allele T of rs77067995 are the risk factors of AMI. Conclusions DSVs in GATA5 promoter can increase susceptibility to AMI. But the mechanism remains to be verified in vivo.

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Functional genetic variants of the GATA4 gene promoter in acute myocardial infarction

Molecular Medicine Reports ◽

10.3892/mmr.2019.9914 ◽

2019 ◽

Author(s):

Jing Chen ◽

Shuai Wang ◽

Shuchao Pang ◽

Yinghua Cui ◽

Bo Yan ◽

...

Keyword(s):

Myocardial Infarction ◽

Acute Myocardial Infarction ◽

Genetic Variants ◽

Gene Promoter

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Functional genetic variants within the SIRT2 gene promoter in acute myocardial infarction

PLoS ONE ◽

10.1371/journal.pone.0176245 ◽

2017 ◽

Vol 12 (4) ◽

pp. e0176245 ◽

Cited By ~ 3

Author(s):

Wentao Yang ◽

Feng Gao ◽

Pei Zhang ◽

Shuchao Pang ◽

Yinghua Cui ◽

...

Keyword(s):

Myocardial Infarction ◽

Acute Myocardial Infarction ◽

Genetic Variants ◽

Gene Promoter ◽

Sirt2 Gene

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TFEB Gene Promoter Variants Effect on Gene Expression in Acute Myocardial Infarction

Frontiers in Cell and Developmental Biology ◽

10.3389/fcell.2021.630279 ◽

2021 ◽

Vol 9 ◽

Author(s):

Jie Zhang ◽

Yexin Zhang ◽

Xiaohui He ◽

Shuai Wang ◽

Shuchao Pang ◽

...

Keyword(s):

Gene Expression ◽

Myocardial Infarction ◽

Acute Myocardial Infarction ◽

Gene Promoter ◽

Nucleotide Polymorphisms ◽

Lysosomal Hydrolases ◽

Mobility Shift ◽

Regulatory Variants ◽

Gene Regulatory ◽

Mobility Shift Assay

Autophagy is involved in many physiological processes. Transcription factor EB (TFEB) is a master regulator of autophagy and coordinates the expression of autophagic proteins, lysosomal hydrolases, and lysosomal membrane proteins. Though autophagy has been implicated in several human diseases, little is known regarding TFEB gene expression and regulation in the process. Since dysfunctional autophagy plays critical roles in acute myocardial infarction (AMI), dysregulated TFEB gene expression may be associated with AMI by regulating autophagy. In this study, the TFEB gene promoter was genetically and functionally analyzed in AMI patients (n = 352) and ethnic-matched controls (n = 337). A total of fifteen regulatory variants of the TFEB gene, including eight single-nucleotide polymorphisms (SNPs), were identified in this population. Among these, six regulatory variants [g.41737274T>C (rs533895008), g.41737144A>G, g.41736987C > T (rs760293138), g.41736806C > T (rs748537297), g.41736635T > C (rs975050638), and g.41736544C > T] were only identified in AMI patients. These regulatory variants significantly altered the transcriptional activity of the TFEB gene promoter. Further electrophoretic mobility shift assay revealed that three of the variants evidently affected the binding of transcription factors. Therefore, this study identified novel TFEB gene regulatory variants which affect the gene expression. These TFEB gene regulatory variants may contribute to AMI development as a rare risk factor.

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The nuclear orphan receptors COUP-TFII and Ear-2 act as silencers of the human oxytocin gene promoter

Journal of Molecular Endocrinology ◽

10.1677/jme.0.0190163 ◽

1997 ◽

Vol 19 (2) ◽

pp. 163-172 ◽

Cited By ~ 27

Author(s):

K Chu ◽

HH Zingg

Keyword(s):

Receptor Binding ◽

Promoter Activity ◽

Transcriptional Activity ◽

Gene Promoter ◽

Fine Tuning ◽

Orphan Receptor ◽

Expression Vectors ◽

Mobility Shift ◽

Interaction Sites

We have previously shown that COUP-TFII and Ear-2, two members of the nuclear orphan receptor family, are able to repress oestrogen-stimulated transcriptional activity of the human oxytocin (OT) gene promoter by binding to a site that overlaps with the oestrogen response element (ERE) present in the 5' flanking region of the gene. Although most nuclear receptor-mediated transcriptional repression conforms with the paradigm of passive repression and involves competitive binding to an activator site, active repression, i.e. silencing of basal promoter activity, has been observed in a limited number of cases. Here we show by co-transfection experiments using COUP-TFII and Ear-2 expression vectors and reporter constructs containing OT gene promoter fragments linked to the chloramphenicol acetyltransferase gene that both COUP-TFII and Ear-2 are capable of silencing basal OT gene promoter activity by 54 and 75% respectively. 5' Deletion and footprint analyses revealed two areas of functionally important interaction sites: (1) a direct TGACC(T/C) repeat overlapping the ERE and (2) a more promoter-proximal area centred at - 90 containing three imperfect direct repeats (R1-R3) spaced by four nucleotides each. Mutagenesis of reporter constructs as well as electrophoretic mobility-shift assays demonstrated that each of the three proximal repeats R1-R3 contributed to orphan receptor binding and the silencing effect. Inasmuch as the orphan receptor-binding sites are not involved in mediating basal transcriptional activity of the OT gene promoter, the observed effects are best interpreted as active repression or promoter silencing. Moreover, since COUP-TFII and Ear-2 are both co-expressed in OT-expressing uterine epithelial cells, the novel transcriptional effects described here are likely to be of functional importance in the fine-tuning of uterine OT gene expression in vivo.

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