子宫内膜异位症的多肽组学研究

Endometriosis,a common disease in women of reproductive age, has extensive lesions which destroy functions of organs in pelvic and abdominal cavity causing adhesions,pain and infertility. It adversely impairs women' health and quality of life.The pathogenesis of endometriosis is unclear,early diagnosis is difficult,effect of treatment is not satisfactory.It is a medical challenge worldwide. In our previous investigation, we found that in blood urine and perineal fluid of patients with endometriosis there were differently-expressed proteins which could be biomarkers of endometriosis.However, no much attention has bee paid on study of peptidome in body fluid of patients with endometriosis.In our preliminary experiment,we found that there were several differently-expressed peptides in urine and perineal fluid of patients with endometriosis. They might be biomarkers of endometriosis.The present study will systemicly mine the peptides in perineal fluid and urine of patients with endometriosis with further identification using mass spectrometry and functional analysis.In order to screen the endometriosis related peptides, we will using ELISA，Western-blotting and colloidal gold method and further verify by clinical large sample trial.According to the functions of peptides,we will design project and investigate the functions of these peptides in pathogenesis of endometriosis. We also will construct a urine peptide model for diagnosis of endometriosis using bioinformatics and study its value in clinical usage through clinical large sample trial.

子宫内膜异位症（内异症）是育龄妇女的常见病，其病变广泛，破坏盆腹腔器官功能，导致粘连、疼痛和不育，严重影响妇女的健康和生活质量。内异症发病机制不清，临床早期诊断困难，治疗效果不佳，是世界性的医学难题。本课题组前期研究发现，在内异症发病过程中，患者血液、腹水和尿液中有异常表达的蛋白；而目前关于内异症患者体液中多肽组的研究未得到重视。本课题组预实验发现，内异症患者腹水及尿液中有多个异常表达的多肽，可能成为内异症标志性多肽。本研究通过多肽组学方法将系统性挖掘内异症患者血液、腹水及尿液中异常表达的多肽，进行质谱鉴定及结构功能分析；通过ELISA、Western-blotting或者胶体金方法进一步筛选，然后经临床大样本验证，筛选内异症相关多肽；根据内异症相关多肽的功能，设计实验研究其在内异症发病中的作用；通过生物信息学构建内异症体液多肽诊断模型，通过临床大样本研究，探讨其临床应用价值。

子宫内膜异位症影响约10%-15%的生育年龄女性，具体机制不详，缺乏特异性早期诊断标志物。本研究首先采用基于TMT标记的高通量蛋白质组相对定量方法对卵巢子宫内膜异位症囊肿、卵巢良性肿瘤、卵巢恶性肿瘤及正常人群的尿液差异蛋白质进行高通量筛选。生物信息学分析的结果显示子宫内膜异位症在高迁移族蛋白B1（HMGB1）通路、CD40通路、Lymphotoxin β Receptor 通路中明显正向激活。使用靶向蛋白质组学定量技术平行反应监测（PRM）对筛选的差异蛋白结果进行验证。本研究共筛选并验证出子宫内膜异位症差异蛋白25个，其中18个具有诊断价值（AUC＞0.75），组蛋白4（H4）诊断效率最高（AUC 0.851）。构建出四个蛋白组合H4、RPN1、ARF3、MYH10的诊断效率最高（AUC 0.863）。研究结果提示慢性炎症在内异症的发生发展中可能起着重要作用。此外构建了针对卵巢子宫内膜异位症囊肿的诊断试剂盒，诊断效率可（AUC 0.764），具有一定使用价值。本研究第一次将TMT-PRM技术流程应用于子宫内膜异位症研究，也是首次发现组蛋白在子宫内膜异位症病人体液中含量升高，这将有助于未来子宫内膜异位症发病机制和治疗靶点的研究。