Kurz Carolin, Hakimi Maani, Kloor Matthias, Grond-Ginsbach Caspar, Gross-Weissmann Marie-Luise, Böckler Dittmar, von Knebel Doeberitz Magnus, Dihlmann Susanne
Department of Neurology, Technical University Munich, Munich, Germany.
Department of Vascular and Endovascular Surgery, University Hospital Heidelberg, Heidelberg, Germany.
Mol Med. 2015 Jun 9;21(1):479-86. doi: 10.2119/molmed.2014.00258.
Somatic DNA alterations are known to occur in atherosclerotic carotid artery lesions; however, their significance is unknown. The accumulation of microsatellite mutations in coding DNA regions may reflect a deficiency of the DNA mismatch repair (MMR) system. Alternatively, accumulation of these coding microsatellite mutations may indicate that they contribute to the pathology. To discriminate between these two possibilities, we compared the mutation frequencies in coding microsatellites (likely functionally relevant) with those in noncoding microsatellites (likely neutral). Genomic DNA was isolated from carotid endarterectomy (CEA) specimens of 26 patients undergoing carotid surgery and from 15 nonatherosclerotic control arteries. Samples were analyzed by DNA fragment analysis for instability at three noncoding (BAT25, BAT26, CAT25) and five coding (AIM2, ACVR2, BAX, CASP5, TGFBR2) microsatellite loci, with proven validity for detection of microsatellite instability in neoplasms. We found an increased frequency of coding microsatellite mutations in CEA specimens compared with control specimens (34.6 versus 0%; p = 0.0013). Five CEA specimens exhibited more than one frameshift mutation, and ACVR2 and CASP5 were affected most frequently (5/26 and 6/26). Moreover, the rate of coding microsatellite alterations (15/130) differed significantly from that of noncoding alterations (0/78) in CEA specimens (p = 0.0013). In control arteries, no microsatellite alterations were observed, neither in coding nor in noncoding microsatellite loci. In conclusion, the specific accumulation of coding mutations suggests that these mutations play a role in the pathogenesis of atherosclerotic carotid lesions, since the absence of mutations in noncoding microsatellites argues against general microsatellite instability, reflecting MMR deficiency.
已知体细胞DNA改变会发生在动脉粥样硬化性颈动脉病变中;然而,其意义尚不清楚。编码DNA区域中微卫星突变的积累可能反映了DNA错配修复(MMR)系统的缺陷。或者,这些编码微卫星突变的积累可能表明它们与病理过程有关。为了区分这两种可能性,我们比较了编码微卫星(可能具有功能相关性)和非编码微卫星(可能是中性的)中的突变频率。从26例接受颈动脉手术的患者的颈动脉内膜切除术(CEA)标本以及15条非动脉粥样硬化对照动脉中分离基因组DNA。通过DNA片段分析对样本进行分析,检测三个非编码(BAT25、BAT26、CAT25)和五个编码(AIM2、ACVR2、BAX、CASP5、TGFBR2)微卫星位点的不稳定性,这些位点已被证明可有效检测肿瘤中的微卫星不稳定性。我们发现,与对照标本相比,CEA标本中编码微卫星突变的频率增加(34.6%对0%;p = 0.0013)。5例CEA标本表现出不止一个移码突变,ACVR2和CASP5受影响最频繁(分别为5/26和6/26)。此外,CEA标本中编码微卫星改变的发生率(15/130)与非编码改变的发生率(0/78)有显著差异(p = 0.0013)。在对照动脉中,无论是编码微卫星位点还是非编码微卫星位点,均未观察到微卫星改变。总之,编码突变的特异性积累表明这些突变在动脉粥样硬化性颈动脉病变的发病机制中起作用,因为非编码微卫星中无突变表明不存在一般的微卫星不稳定性,这反映了MMR缺陷。