Sasse A, Wallich M, Ding Z, Goedecke A, Schrader J
Institut für Herz- und Kreislaufphysiologie, Heinrich-Heine-University Duesseldorf, Universitaetsstr. 1, 40225 Duesseldorf, Germany.
J Gene Med. 2003 Oct;5(10):876-882. doi: 10.1002/jgm.411.
Adenoviral vectors are widely used as gene-transfer vehicles in experimental and clinical studies. Since virus incorporation and transfection efficacy depend to a large extent on the concentration of the coxsackie-and-adenovirus (CAR) receptor on target cells the aim of this study was to quantify the CAR-receptor concentration in various human cardiomyopathies.
After RNA isolation from myocardial biopsies obtained during surgical procedures, cDNA was generated by reverse transcription. The relative RNA content was analyzed by quantitative PCR using glyceraldehydes-3-phosphate dehydrogenase (GAPDH) as a standard reference. The cardiomyopathies (CM) analyzed were categorized according to their etiology in dilated CM (DCM, n = 28), ischemic CM (ICM, n = 52), CM in mitral valve disease (MVCM, n = 32) and aortic valve disease (AVCM, n = 32). Data were related to non-cardiomyopathic tissue from donor hearts (non-CM, n = 64).
Compared with non-CM hearts DCM showed a 34-fold (+/-5.4) increase in CAR mRNA concentration, in ICM CAR mRNA was elevated by a factor of 12 (+/-4.3), in MVCM by 27 (+/-7) and AVCM by factor 47 (+/-9.3) (ANOVA p < 0.001). Compared with the expression in rat hearts CAR levels were found to be similar to those in human ICM.
These results show that cardiomyopathies associated with heart failure transcribe substantially higher levels - on average by a factor of 30 - of CAR-mRNA than non-failing control hearts. Myocardial gene transfer using adenoviral vectors should therefore be facilitated in human cardiomyopathies and may present a promising approach for therapeutic interventions.
腺病毒载体在实验和临床研究中被广泛用作基因转移载体。由于病毒整合和转染效率在很大程度上取决于靶细胞上柯萨奇病毒和腺病毒（CAR）受体的浓度，本研究的目的是量化各种人类心肌病中CAR受体的浓度。
在手术过程中从心肌活检组织中分离RNA后，通过逆转录生成cDNA。使用甘油醛-3-磷酸脱氢酶（GAPDH）作为标准参照，通过定量PCR分析相对RNA含量。所分析的心肌病（CM）根据其病因分类为扩张型CM（DCM，n = 28）、缺血性CM（ICM，n = 52）、二尖瓣疾病中的CM（MVCM，n = 32）和主动脉瓣疾病中的CM（AVCM，n = 32）。数据与来自供体心脏的非心肌病组织（非CM，n = 64）相关。
与非CM心脏相比，DCM中CAR mRNA浓度增加了34倍（±5.4），ICM中CAR mRNA升高了12倍（±4.3），MVCM中升高了27倍（±7），AVCM中升高了47倍（±9.3）（方差分析p < 0.001）。与大鼠心脏中的表达相比，发现CAR水平与人ICM中的相似。
这些结果表明，与心力衰竭相关的心肌病转录的CAR-mRNA水平比无心力衰竭的对照心脏平均高30倍。因此，在人类心肌病中使用腺病毒载体进行心肌基因转移应更容易实现，并且可能是一种有前途的治疗干预方法。
