Lakatos László, Lakatos Péter László, Willheim-Polli Claudia, Reinisch Walter, Ferenci Péter, Tulassay Zsolt, Molnár Tamás, Kovács Agota, Papp János, Szalay Ferenc
Csolnoky Ferenc Kórház, I. Sz. Belgyógyászati Osztály, Veszprém.
Orv Hetil. 2004 Jul 4;145(27):1403-11.
Recently mutations of the NOD2/CARD15 gene were identified as genetic markers for Crohn's disease (CD). It has also been suggested that the presence of mutation may influence the phenotype of the disease. The aim of this study was to determine the frequency of common NOD2/CARD15 mutations in Hungarian Crohn's patients.
DNA was obtained from 142 pts with Crohn's disease (70 male and 72 female, mean age: 36.2 yrs) and of 115 healthy subjects. Clinical data were obtained by filling in a questionnaire by the physician. DNA was screened for possible mutations by denaturing HPLC (WAVE Nucleic acid fragment analysis systems, Cheshire, UK) using published primers (Lesage et al, Am J Hum Gen 2002) for SNP's 8, 12, and 13. Mutations were then confirmed by direct sequencing on a ABI Prism 310 Genetic Analyzer (Perkin Elmer; Norwalk, USA).
Mutations of NOD2/CARD15 were significantly more frequent in patients (n = 42, 29.6%) than in controls (11.3%, p = 0.0007, OR = 3.3, 95% CI = 1.7-6.5). R702W and 3020insC mutations were significantly more common in IBD compared to controls (4.3% and 2.6%); 18 patients had the R702W mutation (12.7%, p = 0.001, 13 homozygous, one homozygous for R703C) and 22 the 3020insC (15.9%, p = 0.001, 4 homozygous). 7 patients (4.9%) were heterozygous for the G908R, which was not different from the controls (4.3%). 5 patients were compound heterozygous. The allele frequency of R702W (11.6% vs. 4.3%, p = 0.0026, OR = 5.9, 95% CI: 2.3-14.9) and 3020insC (9.1% vs. 2.6% p = 0.004, OR = 7.6, 95% CI = 2.4-24.0) was significantly higher in Crohn's disease compared to the controls. Age at presentation was not different between carriers and non-carriers (26.7 +/- [SD] 10.3 yrs vs. 27.7 +/- 11.8 yrs). There was a tendency of ileal location to be more common in carriers of the mutation (40.5% vs. 29.0%), while colonic location was less frequent (16.7% vs. 44%). The presence of the mutation did not affect disease behaviour (carrier: inflammatory: 30.9%, stenosing: 26.2%, penetrating: 42.9%, non-carrier: inflammatory: 36%, stenosing: 27%, penetrating: 37%). Among the extraintestinal manifestations arthritis (30.1% vs. 47%, p = 0.05) and primary sclerosing cholangitis (0% vs. 9%, p = 0.047%) was significantly less frequent in carriers of the mutation.
In concordance with European data we found a high number of NOD2/CARD15 R702W and 3020insC mutations in Hungarian patients with Crohn's disease. The G908R mutation was uncommon in Hungarian Crohn's patients. The presence of the mutation was associated with ileal but not with fibrostenosing disease and extraintestinal manifestations were less common in carriers of the mutation.
最近，NOD2/CARD15基因的突变被确定为克罗恩病（CD）的遗传标记。也有人提出，突变的存在可能会影响疾病的表型。本研究的目的是确定匈牙利克罗恩病患者中常见NOD2/CARD15突变的频率。
从142例克罗恩病患者（70例男性和72例女性，平均年龄：36.2岁）和115名健康受试者中获取DNA。临床数据由医生通过填写问卷获得。使用已发表的针对单核苷酸多态性8、12和13的引物（Lesage等人，《美国人类遗传学杂志》2002年），通过变性高效液相色谱法（WAVE核酸片段分析系统，英国柴郡）筛选DNA中可能的突变。然后在ABI Prism 310遗传分析仪（珀金埃尔默公司；美国诺沃克）上通过直接测序确认突变。
NOD2/CARD15突变在患者中（n = 42，29.6%）比在对照组中（11.3%，p = 0.0007，OR = 3.3，95%可信区间 = 1.7 - 6.5）明显更频繁。与对照组相比，R702W和3020insC突变在炎症性肠病中明显更常见（4.3%和2.6%）；18例患者有R702W突变（12.7%，p = 0.001，13例纯合子，1例R703C纯合子），22例有3020insC突变（15.9%，p = 0.001，4例纯合子）。7例患者（4.9%）为G908R杂合子，与对照组（4.3%）无差异。5例患者为复合杂合子。R702W（11.6%对4.3%，p = 0.0026，OR = 5.9，95%可信区间：2.3 - 14.9）和3020insC（9.1%对2.6%，p = 0.004，OR = 7.6，95%可信区间 = 2.4 - 24.0）的等位基因频率在克罗恩病患者中比对照组明显更高。发病时的年龄在携带者和非携带者之间无差异（26.7 ± [标准差] 10.3岁对27.7 ± 11.8岁）。突变携带者中回肠部位的倾向更常见（40.5%对29.0%），而结肠部位则较少见（16.7%对44%）。突变的存在不影响疾病行为（携带者：炎症性：30.9%，狭窄性：26.2%，穿透性：42.9%，非携带者：炎症性：36%，狭窄性：27%，穿透性：37%）。在肠外表现中，关节炎（30.1%对47%，p = 0.05）和原发性硬化性胆管炎（0%对9%，p = 0.047%）在突变携带者中明显较少见。
与欧洲数据一致，我们发现匈牙利克罗恩病患者中有大量NOD2/CARD15的R702W和3020insC突变。G908R突变在匈牙利克罗恩病患者中不常见。突变的存在与回肠病变相关，但与纤维狭窄性疾病无关，且肠外表现在突变携带者中较少见。
