基于家系的腰椎间盘突出症患者强效基因筛选策略
Potent gene screening strategy based on family-based patients with lumbar discherniation

摘要：目的 对一个椎间盘退变疾病高发家系中4名椎间盘突出患者进行外显子基因测序，寻找此家系中可能导致椎间盘退变的强效致病基因，为研究椎间盘疾病的发病原因以及致病机制提供可能的帮助。方法 收集西安交通大学第一附属医院、西安医学院第一附属医院、西安唐城医院自2010年1月至2016年12月之间就诊的腰椎间盘突出患者，对其进行详细的病史采集及家系调查，从中找到椎间盘退变疾病高发的1个家系。在此家系中，20～50岁成员中有4人为腰椎间盘突出症患者。利用外显子测序技术对这4例患者进行外显子基因分析，检测4例患者的共同外显子突变位点。结果 最终在4个样本中检测出IGFBP6的共同突变Chr12:g.53494591T>C。Sanger法对其进行家系内验证，IGFBP6在4例患者中皆表现为C/T型，而在5例正常对照者中皆表现为T型。最后在正常对照组200例中进行验证，未发现此位点变异。结论 IGFBP6的突变位点c.T430C(p.S144P) (Chr12:g.53494591T>C)可能是导致该家系中椎间盘突出疾病高发的因素；外显子测序技术可以应用于复杂疾病致病基因的发掘。
ABSTRACT: Objective To look for a potent gene that may cause intervertebral disc degeneration in a family with intervertebral disc herniation by exome sequencing four patients with intervertebral disc herniation so as to provide possible help for studies on the causes and mechanisms of intervertebral disc diseases. Methods We collected the serum of patients with lumbar disc herniation treated at The First Affiliated Hospital of Xi’an Jiaotong University, The First Affiliated Hospital of Xi’an Medical College, and Xi’an Tangcheng Hospital from January 2010 to December 2016. A detailed medical history was collected and family survey was conducted to find a family with high incidence of degenerative intervertebral disc disease. Then we found 4 patients aged 20 to 50 years with lumbar disc herniation in this family. Exome sequencing was used to analyze the common exon mutation sites of the 4 patients to find the common mutation sites. Results The common mutation IGFBP6, Chr12:g.53494591T>C, was detected in the 4 samples. The Sanger sequencing performed in-family validation revealed that the IGFBP6 showed a C/T type in all the four patients and a T type in all the five normal controls. Finally, verification was performed in 200 normal controls and no mutation was found in this site. Therefore, the IGFBP6 might be the effector gene that caused intervertebral disc degeneration in this family. Conclusion The mutation site of IGFBP6, c.T430C (p.S144P) (Chr12:g.53494591T>C), may be a factor that contributes to the high incidence of intervertebral disc herniation in this family. We still need to verify the function of the gene in future. Our study has also confirmed that exome sequencing technology can be applied to the detection of disease-causing genes in complex diseases