长链非编码RNA LBX1-AS1在青少年特发性脊柱侧凸发病中的调控机制研究

Adolescent idiopathic scoliosis (AIS) is a structural deformity of the spine estimated to affect millions of children with a prevalence of 2% - 4%. Commonly understood as a complex disease, AIS is believed to be resulted from the interaction among multiple genetic loci as well as certain environmental factors. LBX1 was the only susceptible gene of AIS that can be widely validated in different populations, including Chinese Han, Japanese and the Caucasians. Recently we have completed the first genome-wide association study of AIS in Chinese Han population. Combining the data of discovery stage and 2 replication stages, we found several SNPs significantly associated with AIS, with p value less than 10-8. Among these SNPs, s678741 and rs10883597 are located in long non-coding RNA LBX1-AS1, which is the anti-sense RNA of LBX1 as indicated by the dataset of NCBI. Since previous studies has reported lncRNA revealed by GWAS can probably be functionally implicated in the etiology of the disease, we therefore speculated that LBX1-AS1 can play a role in the etiopathogenesis of AIS through regulation of LBX1. In this project, we plan to investigate the relationship between the expression of LBX1-AS1 and LBX1 in both vertebral body and spinal muscles. Besides, we will pinpoint the functional SNPs that can influence the expression of LBX1-AS1. Finally, luciferase assay is to be performed to verify the regulation of LBX1-AS1 on LBX1. We believe this project can expand our understanding on the etiopathogenesis of AIS.

青少年特发性脊柱侧凸(AIS)是一组原因不明的脊柱三维畸形。遗传因素在AIS的发生过程中所起的作用早已引起了广泛的关注。LBX1基因是首个被在不同种群被广泛验证的AIS疾病易感基因。近期本课题组已完成首个汉族AIS人群的全基因组关联分析研究（GWAS），发现两个与AIS显著相关的单核苷酸多态性位点rs678741和rs10883597均位于LBX1基因附近的长链非编码RNA（lncRNA）LBX1-AS1。查询NCBI数据库表明LBX1-AS1是LBX1基因的反义RNA。参考其他lncRNA被运用于GWAS筛选的易感性位点的功能验证研究，我们推断 LBX1-AS1可能通过调控LBX1的表达从而参与AIS的发病机制。本项目拟观察LBX1-AS1在脊柱组织中对LBX1表达的调控，并分析影响其表达的功能性易感位点，最终在细胞学水平对上述观察结果进行验证。研究成果将有助于进一步理解AIS发病机制。

青少年特发性脊柱侧凸(AIS)是一组原因不明的脊柱三维畸形。遗传因素在AIS的发生过程中所起的作用早已引起了广泛的关注。LBX1基因是首个被在不同种群被广泛验证的AIS疾病易感基因。近期本课题组已完成首个汉族AIS人群的全基因组关联分析研究（GWAS），发现两个与AIS显著相关的单核苷酸多态性位点rs678741和rs10883597均位于LBX1基因附近的长链非编码RNA（lncRNA）LBX1-AS1。查询NCBI数据库表明LBX1-AS1是LBX1基因的反义RNA。参考其他lncRNA被运用于GWAS筛选的易感性位点的功能验证研究，我们推断 LBX1-AS1可能通过调控LBX1的表达从而参与AIS的发病机制。本项目初步探索了LBX1-AS1参与AIS发生的分子机制。我们发发现在前期GWAS研究发现的易感位点rs678741对LBX1-AS1表达有显著调控能力，表现为患者LBX1蛋白水平较对照组显著下调，同时风险基因型TT对应的LBX1表达较其他基因型亦显著下调。报告基因和EMSA表明等位基因T可通过影响转录因子EST-1的结合从而影响启动子转录活性。作为LBX1-AS1的靶基因，LBX1表达与患者II型肌纤维存在显著相关性。通过提纯和培养患者肌卫星细胞，本研究证实了过表达LBX1-AS1可影响肌细胞增殖和分化。综上，LBX1-AS1可能通过影响AIS患者肌肉组织发育从而在疾病发生中起一定作用。其表达水平可能与疾病进程相关。这为患者诊疗提供了新的线索。