汉族人群中拉莫三嗪诱导HLA-A*2402、B*1502结合肽在重症皮肤型药物不良反应中的作用机制

Lamotrigine (LTG) is a commonly used antiepileptic drug. However, the use of LTG is limited because of its cutaneous adverse drug reactions especially Stevens–Johnson syndrome (SJS) and toxic epidermal necrolysis (TEN) which had a mortality up to 30%. Therefore, preventing SJS/TEN and improving patient safety is an important goal of anticonvulsant therapy. Recently, our study suggest that both HLA-A*2402 and HLA-B*1502 are associated with LTG-SJS/TEN in Han Chinese. The pathogenesis of SJS/TEN remain unknown. Available evidence suggests that drug hypersensitivity is mediated by adaptive immunity, which involves drugs, HLA molecules, and peptides. In this study, we will construct HLA-A*2402 and HLA- B*1502 overexpression Hmy2.CIR cell lines by lentivirus packing. Hmy2.CIR cell lines were cultured in the presence or absence of LTG, HLA binding peptide was examined by using mass spectrometry and bioinformatics software. We investigated the effects of HLA-A*2402、HLA-B*1502 bound peptides induced by LTG in the severe cutaneous adverse drug reactions in Han Chinese. The result will be benefited for screening for risk factors before prescript LTG as well as providing the therapeutic target for the treatment of SJS/TEN.

拉莫三嗪(LTG) 是临床常用的抗癫痫药物(AED)，其导致的Stevens-Johnson综合症（SJS）、中毒性表皮坏死松解症（TEN）是重症皮肤型药物不良反应。SJS/TEN的致残致死率高达30%，避免其发生意义重大。本课题前期研究表明人类白细胞抗原HLA-Ⅰ型基因A*2402、 B*1502与LTG-SJS/TEN 相关。目前SJS/TEN发病机制未明，可能与药物、特异型HLA、结合肽等有关。本项目拟通过克隆HLA-A*2402、 B*1502基因片段，慢病毒包装后转入无HLA-Ⅰ型基因表达的Hmy2.CIR细胞系，通过质谱检测、生物信息学等方法，了解加LTG前后HLA结合肽的差异，从而获得LTG诱导的HLA-A*2402、 B*1502结合肽在SJS/TEN发生中的作用。为临床用药前开展HLA风险基因的筛查、推动个体化医疗提供理论依据，为开发防治SJS/TEN的药物提供实验依据。

芳香族抗癫痫药物（AEDs）导致的皮肤不良反应（cADRS）常导致患者停药或撤药，避免其发生意义重大。拉莫三嗪(LTG)是新一代的芳香族AEDs，前期研究显示HLA-A*2402、 B*1502是芳香族AEDs导致Stevens-Johnson综合症（SJS）、中毒性表皮坏死松解症（TEN）的危险基因型，本项目通过多中心大样本的研究验证了汉族人群中HLA-B*1502、HLA-A*2402基因是芳香族AEDs导致重症皮肤不良反应的风险基因型；发现HLA-B*1301是AEDs导致皮肤型交叉反应的风险基因；进一步克隆HLA-A*2402、 B*1502基因片段，慢病毒包装后转入无HLA-Ⅰ型基因表达的Hmy2.CIR细胞系，通过质谱检测、生物信息学等方法发现MARCH4蛋白、冠蛋白（CORONIN 1A）、微管蛋白β4 (TUBB4B)可能参与LTG-SJS的发病机制。本项目为临床用药前筛查HLA风险基因型，推动精准医疗提供理论依据，为开发防治皮肤药物不良反应的靶点药物提供实验依据。