Expression profiling of cardiac genes in human hypertrophic cardiomyopathy: Insight into the pathogenesis of phenotypes
Expression profiling of cardiac genes in human hypertrophic cardiomyopathy: Insight into the pathogenesis of phenotypes
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DOI：
10.1016/s0735-1097(01)01509-1
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发表时间：
2001-10-01
影响因子：
24
通讯作者：
Marian, AJ
中科院分区：
文献类型：
--
作者：
Lim, DS;Roberts, R;Marian, AJ
本研究的目的是鉴定肥厚型心肌病（HCM）患者心脏中上调的基因。背景肥厚型心肌病是由收缩性肌节蛋白突变引起的遗传性疾病。HCM临床和病理表型多样性的分子基础尚不清楚。方法采用聚合酶链反应-选择性互补DNA差减技术，在正常心脏和HCM心脏之间进行差减，用Southern印迹法筛选差减文库。我们对差异表达的克隆进行测序，并进行北方印迹检测增加的表达水平。结果我们筛选了288个独立的克隆，其中76个克隆的信号强度增加不到两倍，被认为是上调的。序列分析鉴定了36个基因，包括编码压力超负荷诱导的（“继发性”）心肌肥大的标志物、细胞骨架蛋白、蛋白质合成、氧化还原系统、离子通道和功能未知的基因。北方印迹证实了骨骼肌α-肌动蛋白（ACTA 1）、肌球蛋白轻链2a（MLC 2a）、GTP结合蛋白GS-α亚基（GNAS 1）、NADH泛醌氧化还原酶（NDUFB 10）、电压依赖性阴离子通道1（VDAC 1）、四个半LIM结构域蛋白1（FHL 1）的表达增加。（也称为SLIM 1）、肌氨酸（sarcosin）和热休克70 kD蛋白8（HSPA 8）的蛋白质水平降低不到两倍。ACTA 1，MLC 2a和GNAS 1的表达水平增加6个额外的和FHL 1在4个额外的心脏HCM。结论一个不同的阵列的基因上调，在人类HCM患者的心脏，这可以解释临床和病理表型的多样性。继发性肥大的标志物也上调，表明HCM和获得性心脏肥大的途径的共同性。阐明差异表达基因在HCM中的作用可以提供新的治疗靶点。(C)2001年，美国心脏病学会。
OBJECTIVES The goal of this study was to identify genes upregulated in the heart in human patients with hypertrophic cardiomyopathy (HCM).BACKGROUND Hypertrophic cardiomyopathy is a genetic disease caused by mutations in contractile sarcomeric proteins. The molecular basis of diverse clinical and pathologic phenotypes in HCM remains unknown.METHODS We performed polymerase chain reaction-select complementary DNA subtraction between normal hearts and hearts with HCM and screened subtracted libraries by Southern blotting. We sequenced the differentially expressed clones and performed Northern blotting to detect increased expression levels.RESULTS We screened 288 independent clones, and 76 clones had less than twofold increase in the signal, intensity and were considered upregulated. Sequence analysis identified 36 genes including those encoding the markers of pressure overload-induced ("secondary") cardiac hypertrophy, cytoskeletal proteins, protein synthesis, redox system, ion channels and those with unknown function. Northern blotting confirmed increased expression of skeletal muscle alpha-actin (ACTA1), myosin light chain 2a (MLC2a), GTP-binding protein Gs-alpha subunit (GNAS1), NADH ubiquinone oxidoreductase (NDUFB10), voltage-dependent anion channel 1 (VDAC1), four-and-a-half LIM domain protein 1 (FHL1) (also known as SLIM1), sarcosin (SARCOSIN) and heat shock 70kD protein 8 (HSPA8) by less than twofold. Expression levels of ACTA1, MLC2a and GNAS1 were increased in six additional and FHL1 in four additional hearts with HCM.CONCLUSIONS A diverse array of genes is upregulated in the heart in human patients with HCM, which could account for the diversity of clinical and pathologic phenotypes. Markers of secondary hypertrophy are also upregulated, suggesting commonality of pathways involved in HCM and the acquired forms of cardiac hypertrophy. Elucidation of the role of differentially expressed genes in HCM could provide for new therapeutic targets. (C) 2001 by the American College of Cardiology.