先天性心脏病胎儿母体血清中的microRNA表达谱及其诊断意义
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摘要
目的筛选先天性心脏病胎儿的母体血清差异表达microRNA(miRNA),确定先天性心脏病的无创产前诊断候选标志物。方法 2012年1月至2015年12月,在中国医科大学附属盛京医院进行产前超声检查并确诊为胎儿先天性心脏病的50例孕妇为研究对象;选取50例正常孕妇为对照组,孕周、孕妇年龄与先天性心脏病组相匹配。利用miRNA芯片对先天性心脏病胎儿的母体血清中差异表达的miRNA进行高通量筛选,并进一步用实时定量PCR方法在研究组与对照组胎儿母体血清中验证差异表达miRNA。统计学分析采用独立样本t检验及受试者操作特性曲线。结果芯片方法在孕妇血清中共探测到1 033个人类miRNA,研究组与对照组中差异倍数高于2倍以上的miRNA共38个。生物信息学信号通路分析显示最相关的为心肌细胞中的肾上腺素信号通路和AMPK信号通路。4个miRNA(miR-3199、miR-125b-2-3p、miR-4666a-3p和miR-4681)在先天性心脏病组中表达显著低于对照组;3个miRNA (miR-3664-3p、miR-1275和miR-4796-3p)在先天性心脏病组中表达显著高于对照组。结论孕妇血清中7个差异表达的miRNA可作为胎儿先天性心脏病的无创产前诊断标志物,具有潜在的临床价值。
Objective To screen the differential expressed microRNAs(miRNAs) in maternal serum of pregnant women with congenital heart disease(CHD) fetuses and to confirm the potential biomarkers for non-invasive prenatal diagnosis of CHD. Methods From January 2012 to December 2015, 50 pregnant women who were diagnosed as pregnant with CHD fetuses by prenatal ultrasound examination in Shengjing Hospital of China Medical University were recruited. Fifty cases of normal pregnant women whose gestational week and maternal age matched with the CHD group were selected as the control group. The high-throughput screening was used for the differential expressed microRNAs in maternal serum with CHD fetuses by miRNA microarray analysis. The differential expressed microRNAs in maternal serum were further verified by the method of real-time quantitative PCR between study group and control group. Independent sample t-test and receiver operating characteristic(ROC) curves were used for statistical analysis. Results Among the 1 033 miRNAs detected in the maternal serum by miRNA microarray analysis, 38 miRNAs showed at least a 2-fold difference between study group and control group. Bioinformatics signal pathway analysis showed that the top two pathways were adrenergic signaling pathway and AMPK signaling pathway in cardiomyocytes. The expression of 4 miRNAs(miR-3199, miR-125b-2-3p, miR-4666a-3p and miR-4681) in CHD group was significantly lower than that in control group; the expression of 3 miRNAs(miR-3664-3p, miR-1275 and miR-4796-3p) in CHD group was significantly higher than that in control group. Conclusions The 7 differential expressed miRNAs in maternal serum of pregnant women can be used as the new biomarkers for non-invasive prenatal diagnosis of fetal CHD and has latent clinical value.
Objective To screen the differential expressed microRNAs(miRNAs) in maternal serum of pregnant women with congenital heart disease(CHD) fetuses and to confirm the potential biomarkers for non-invasive prenatal diagnosis of CHD. Methods From January 2012 to December 2015, 50 pregnant women who were diagnosed as pregnant with CHD fetuses by prenatal ultrasound examination in Shengjing Hospital of China Medical University were recruited. Fifty cases of normal pregnant women whose gestational week and maternal age matched with the CHD group were selected as the control group. The high-throughput screening was used for the differential expressed microRNAs in maternal serum with CHD fetuses by miRNA microarray analysis. The differential expressed microRNAs in maternal serum were further verified by the method of real-time quantitative PCR between study group and control group. Independent sample t-test and receiver operating characteristic(ROC) curves were used for statistical analysis. Results Among the 1 033 miRNAs detected in the maternal serum by miRNA microarray analysis, 38 miRNAs showed at least a 2-fold difference between study group and control group. Bioinformatics signal pathway analysis showed that the top two pathways were adrenergic signaling pathway and AMPK signaling pathway in cardiomyocytes. The expression of 4 miRNAs(miR-3199, miR-125b-2-3p, miR-4666a-3p and miR-4681) in CHD group was significantly lower than that in control group; the expression of 3 miRNAs(miR-3664-3p, miR-1275 and miR-4796-3p) in CHD group was significantly higher than that in control group. Conclusions The 7 differential expressed miRNAs in maternal serum of pregnant women can be used as the new biomarkers for non-invasive prenatal diagnosis of fetal CHD and has latent clinical value.
引文
[1]Marelli AJ,Ionescu-Ittu R,Mackie AS,et al.Lifetime prevalence of congenital heart disease in the general population from 2000 to 2010[J].Circulation,2014,130(9):749-756.
[2]Tennant PW,Pearce MS,Bythell M,et al.20-year survival of children born with congenital anomalies:a population-based study[J].Lancet,2010,375(9715):649-656.
[3]van Velzen CL,Clur SA,Rijlaarsdam ME,et al.Prenatal detection of congenital heart disease--results of a national screening programme[J].BJOG,2016,123(3):400-407.
[4]Xu P,Zhao Y,Liu M,et al.Variations of microRNAs in human placentas and plasma from preeclamptic pregnancy[J].Hypertension,2014,63(6):1276-1284.
[5]Higashijima A,Miura K,Mishima H,et al.Characterization of placenta-specific microRNAs in fetal growth restriction pregnancy[J].Prenat Diagn,2013,33(3):214-222.
[6]Ventura W,Koide K,Hori K,et al.Placental expression of microRNA-17 and-19b is down-regulated in early pregnancy loss[J].Eur J Obstet Gynecol Reprod Biol,2013,169(1):28-32.
[7]Gu H,Li H,Zhang L,et al.Diagnostic role of microRNAexpression profile in the serum of pregnant women with fetuses with neural tube defects[J].J Neurochem,2012,122(3):641-649.
[8]Takahashi RU,Prieto-Vila M,Hironaka A,et al.The role of extracellular vesicle microRNAs in cancer biology[J].Clin Chem Lab Med,2017,55(5):648-656.
[9]Ouyang Y,Mouillet JF,Coyne CB,et al.Placenta-specific microRNAs in exosomes-good things come in nano-packages[J].Placenta,2014,35(Suppl):S69-73.
[10]Miura K,Miura S,Yamasaki K,et al.Identification of pregnancy-associated microRNAs in maternal plasma[J].Clin Chem,2010,56(11):1767-1771.
[11]Morisaki S,Miura K,Higashijima A,et al.Effect of labor on plasma concentrations and postpartum clearance of cell-free,pregnancy-associated,placenta-specific microRNAs[J].Prenat Diagn,2015,35(1):44-50.
[12]Tsochandaridis M,Nasca L,Toga C,et al.Circulating microRNAs as clinical biomarkers in the predictions of pregnancy complications[J].Biomed Res Int,2015,2015:1-8.
[13]Mouillet JF,Ouyang Y,Coyne CB,et al.MicroRNAs in placental health and disease[J].Am J Obstet Gynecol,2015,213(4 Suppl):S163-172.
[14]Lycoudi A,Mavreli D,Mavrou A,et al.miRNAs in pregnancy-related complications[J].Expert Rev Mol Diagn,2015,15(8):999-1010.
[15]Xu G,Cui Y,Jia Z,et al.The values of coronary circulating miRNAs in patients with atrial fibrillation[J].PLoS One,2016,11(11):e0166235.
[16]Huan T,Rong J,Tanriverdi K,et al.Dissecting the roles of microRNAs in coronary heart disease via integrative genomic analyses[J].Arterioscler Thromb Vasc Biol,2015,35(4):1011-1021.
[17]Sun LS,Du F,Quaegebeur JM.Right ventricular infundibular beta-adrenoceptor complex in tetralogy of fallot patients[J].Pediatr Res,1997,42(1):12-16.
[18]袁正伟.先天畸形的胚胎早期诊断与治疗新进展[J].发育医学电子杂志,2018,6(2):69-73.
[19]Nikolaev VO,Moshkov A,Lyon AR,et al.Beta2-adrenergic receptor redistribution in heart failure changes cAMPcompartmentation[J].Science,2010,327(5973):1653-1657.
[20]Kozlik-Feldmann R,Kramer HH,Wicht H,et al.Distribution of myocardial beta-adrenoceptor subtypes and coupling to the adenylate cyclase in children with congenital heart disease and implications for treatment[J].J Clin Pharmacol,1993,33(7):588-595.
[2]Tennant PW,Pearce MS,Bythell M,et al.20-year survival of children born with congenital anomalies:a population-based study[J].Lancet,2010,375(9715):649-656.
[3]van Velzen CL,Clur SA,Rijlaarsdam ME,et al.Prenatal detection of congenital heart disease--results of a national screening programme[J].BJOG,2016,123(3):400-407.
[4]Xu P,Zhao Y,Liu M,et al.Variations of microRNAs in human placentas and plasma from preeclamptic pregnancy[J].Hypertension,2014,63(6):1276-1284.
[5]Higashijima A,Miura K,Mishima H,et al.Characterization of placenta-specific microRNAs in fetal growth restriction pregnancy[J].Prenat Diagn,2013,33(3):214-222.
[6]Ventura W,Koide K,Hori K,et al.Placental expression of microRNA-17 and-19b is down-regulated in early pregnancy loss[J].Eur J Obstet Gynecol Reprod Biol,2013,169(1):28-32.
[7]Gu H,Li H,Zhang L,et al.Diagnostic role of microRNAexpression profile in the serum of pregnant women with fetuses with neural tube defects[J].J Neurochem,2012,122(3):641-649.
[8]Takahashi RU,Prieto-Vila M,Hironaka A,et al.The role of extracellular vesicle microRNAs in cancer biology[J].Clin Chem Lab Med,2017,55(5):648-656.
[9]Ouyang Y,Mouillet JF,Coyne CB,et al.Placenta-specific microRNAs in exosomes-good things come in nano-packages[J].Placenta,2014,35(Suppl):S69-73.
[10]Miura K,Miura S,Yamasaki K,et al.Identification of pregnancy-associated microRNAs in maternal plasma[J].Clin Chem,2010,56(11):1767-1771.
[11]Morisaki S,Miura K,Higashijima A,et al.Effect of labor on plasma concentrations and postpartum clearance of cell-free,pregnancy-associated,placenta-specific microRNAs[J].Prenat Diagn,2015,35(1):44-50.
[12]Tsochandaridis M,Nasca L,Toga C,et al.Circulating microRNAs as clinical biomarkers in the predictions of pregnancy complications[J].Biomed Res Int,2015,2015:1-8.
[13]Mouillet JF,Ouyang Y,Coyne CB,et al.MicroRNAs in placental health and disease[J].Am J Obstet Gynecol,2015,213(4 Suppl):S163-172.
[14]Lycoudi A,Mavreli D,Mavrou A,et al.miRNAs in pregnancy-related complications[J].Expert Rev Mol Diagn,2015,15(8):999-1010.
[15]Xu G,Cui Y,Jia Z,et al.The values of coronary circulating miRNAs in patients with atrial fibrillation[J].PLoS One,2016,11(11):e0166235.
[16]Huan T,Rong J,Tanriverdi K,et al.Dissecting the roles of microRNAs in coronary heart disease via integrative genomic analyses[J].Arterioscler Thromb Vasc Biol,2015,35(4):1011-1021.
[17]Sun LS,Du F,Quaegebeur JM.Right ventricular infundibular beta-adrenoceptor complex in tetralogy of fallot patients[J].Pediatr Res,1997,42(1):12-16.
[18]袁正伟.先天畸形的胚胎早期诊断与治疗新进展[J].发育医学电子杂志,2018,6(2):69-73.
[19]Nikolaev VO,Moshkov A,Lyon AR,et al.Beta2-adrenergic receptor redistribution in heart failure changes cAMPcompartmentation[J].Science,2010,327(5973):1653-1657.
[20]Kozlik-Feldmann R,Kramer HH,Wicht H,et al.Distribution of myocardial beta-adrenoceptor subtypes and coupling to the adenylate cyclase in children with congenital heart disease and implications for treatment[J].J Clin Pharmacol,1993,33(7):588-595.