4q D4Z4区域甲基化检测在肩腓综合征鉴别诊断中的价值研究  

Methylation of the 4q D4Z4 Repeat Contributes to Diagnosis and Differential Diagnosis of Scapuloperoneal Muscular Dystrophy

作　　者：夏邢宇 施天仿 成纳川 刘逸奇 岳冬曰[5] 胡超平 王宁宁 焦可馨 孙翀 宋捷[1,2,3] 奚剑英 林洁[1,2,3] 杜坤钊 朱慧[8] 朱雯华 XIA Xing-yu;SHI Tian-fang;CHEN Na-chuan;LIU Yi-qi;YUE Dong-yue;HU Chao-ping;WANG Ning-ning;JAO Ke-xin;SUN Chong;SONG Jie;XI Jian-ying;LIN Jie;DU Kan-zhao;ZHU Hui;ZHU Wen-hua(Department.7.of Neurology,Huashan Hospital,Fudan University,Shanghai 200040,China;National Center for Neurological Diseases,Shanghai 200040,China;Rare Disease Center,Huashan Hospital,Shanghai 200040,China;Shanghai Amplicongene Co.,Ltd,Shanghai 201114,China;Jing'an District Center Hospital of Shanghai,Shanghai 200040,China;Department of Neurology,Children's Hospital of Fudan University,Shanghai 201100,China;Institute for Translational Brain Research Fudan University,Shanghai 200032,China;Department of Cardiology,Huashan Hospital,Fudan University,Shanghai 200040,China)

机构地区：[1]复旦大学附属华山医院神经内科,上海200040 [2]国家神经疾病中心,上海200040 [3]复旦大学附属华山医院罕见病中心,上海200040 [4]上海昂朴医疗科技有限公司,上海201114 [5]复旦大学附属静安区中心医院,上海200040 [6]复旦大学附属儿科医院神经内科,上海201100 [7]复旦大学脑科学转化研究所,上海200032 [8]复旦大学附属华山医院心内科,上海200040

出　　处：《中国临床神经科学》2023年第6期605-613,共9页Chinese Journal of Clinical Neurosciences

基　　金：国家自然科学基金项目(编号:81801242,8217052229);上海市2020年度“科技创新行动计划”生物医药科技支撑专项课题(编号:20S31904200)。

摘　　要：目的 探讨检测4q D4Z4末端甲基化水平在肩腓综合征(SPMD)诊断和鉴别诊断流程中的应用价值。方法纳入60例临床诊断SPMD的患者设为SPMD组、9例家系成员设为家系成员组以及14例其他肌病患者设为对照组,采用亚硫酸盐测序(BSS)结合二代测序对其4qA D4Z4末端D4Z4甲基化水平进行检测和分析。结果 SPMD组患者的平均末端甲基化水平为(21.42±3.98)%。对照组平均末端甲基化水平为(64.78±5.96)%。选定其99%CI的下界值56.48%作为判定阈值,有3例高于此阈值,其余57例均有不同程度甲基化水平下降。54例经基因确诊为面肩肱型肌营养不良(FSHD)1型患者中,甲基化水平不同程度下降53例;D4Z4检测重复次数≥11次的6例患者中,4例甲基化水平降低,其中2例经全基因组测序确诊为FSHD 2型(SMCHD1基因突变)。结论 甲基化筛查有助于完善SPMD的诊断流程,对于甲基化水平降低的患者可优先进行FSHD 1型的分子检测进一步明确诊断。Aim To explore the utility of detecting 4qA D4Z4 distal methylation levels in the diagnosis and differential diagnosis process of scapuloperoneal muscular dystrophy(SPMD).Methods The study included 60 patients clinically diagnosed SPMD, 9 family members, and 14 controls with other muscular diseases. The bisulfite sequencing combined with next-generation sequencing was employed to assess and analyze the distal D4Z4 methylation levels within the 4qA D4Z4. Results The average distal methylation level in the 60 SPMD patients was(21.42±3.98)%. In the 14 non-SPMD patients, it was(64.78±5.96)%. 56.48% of the lower boundary of the 99% confidence interval was selected to determine the threshold. Among all the patients, 3 exceeded this threshold, while the remaining 57 exhibited varying degrees of decreased methylation levels. Among the 54 patients genetically confirmed as facioscapulohumeral muscular dystrophy(FSHD) type 1, 53 displayed varying degrees of decreased methylation. Among the 6 patients with D4Z4 repeat numbers ≥11, 4 showed decreased methylation levels. Additionally, two were confirmed as FSHD type 2(SMCHD1 mutation) through whole-genome sequencing. Conclusion Methylation screening contributes to refining the diagnostic process of SPMD. Patients with decreased methylation levels should be prioritized for further molecular testing to definitively diagnose FSHD type 1.

关 键 词：肩腓综合征 面肩肱型肌营养不良症 D4Z4 甲基化 分子诊断技术 

分 类 号：R742[医药卫生—神经病学与精神病学]