用于银屑病治疗的甲氨蝶呤药物蛋白质组学研究

发布时间：2018-04-22 05:33

本文选题：蛋白质组学 + 寻常型银屑病　；参考：《中国科学院大学(中国科学院上海药物研究所)》2017年硕士论文

【摘要】：银屑病是一种长期的慢性免疫炎症性皮肤病,易复发,难根治。银屑病分为寻常型银屑病、脓疱型银屑病、红皮病型银屑病和关节病型银屑病四种类型,其中,寻常型银屑病是临床中最常见的一种类型,在银屑病中占几乎90%的比重。甲氨蝶呤(MTX)是1971年由美国FDA批准的可用于治疗严重的银屑病的药物。甲氨蝶呤对于患有严重银屑病的患者起效快,但是不能防止复发,部分患者用药后无疗效,并且因其抑制二氢叶酸还原酶来抑制核酸代谢而有严重的副作用,故需要在临床上选择适合的银屑病患者人群进行甲氨蝶呤的用药治疗。随着质谱技术的发展,使得高灵敏度和超高分辨率的质谱技术和更优化的组学方法能够应用于临床样本的研究,推动个体化治疗的进程。本工作运用基于化学标记的定量蛋白质组学方法来研究服用甲氨蝶呤后有无显著疗效患者的外周血单个核细胞(PBMC)的蛋白表达谱,结合临床数据,以期望找到与甲氨蝶呤敏感性和疗效相关的生物标志物,从而对敏感患者人群进行区分,可用于银屑病患者的个体化治疗。我们选取4例甲氨蝶呤有效治疗的患者(治疗0周和8周)、4例甲氨蝶呤无效治疗的患者(治疗0周和8周)和4例正常对照组的共20个外周血单个核细胞样本,采用了体外稳定同位素化学(iTRAQ)标记策略来进行定量蛋白质组学分析。一共鉴定到3397个蛋白,其中3190个蛋白具有定量信息。通过生物信息分析,我们发现一组蛋白在甲氨蝶呤治疗效果好的患者中治疗前后的表达水平发生显著改变,且在甲氨蝶呤治疗后表达恢复到正常水平。我们对其中的ANXA6进行初步的生物学验证,证明甲氨蝶呤治疗效果好的患者,治疗前ANXA6特异性高表达,并在治疗后回复到正常水平。在本工作中,我们首次将药物蛋白质组学技术运用到银屑病患者PBMC细胞中,并发现了与甲氨蝶呤疗效相关的动态蛋白质分子网络。我们的研究结果将为甲氨蝶呤的临床用药予以一定的指导,并对甲氨蝶呤治疗银屑病的药物作用机制研究提供新的线索。
[Abstract]:Psoriasis is a chronic chronic immune inflammatory skin disease, easy to relapse, difficult to cure. Psoriasis is divided into four types: psoriasis vulgaris, pustular psoriasis, psoriasis vulgaris and psoriasis of arthropathy. Among them, psoriasis vulgaris is the most common type, accounting for almost 90% of psoriasis. Methotrexate (MTX) is a drug approved by FDA in 1971 for the treatment of severe psoriasis. Methotrexate acts quickly in patients with severe psoriasis, but does not prevent recurrence. Some patients have no effect after medication, and have serious side effects because of its inhibition of dihydrofolate reductase to inhibit nucleic acid metabolism. Therefore, it is necessary to select suitable patients with psoriasis for methotrexate therapy. With the development of mass spectrometry technology, high sensitivity and ultra-high resolution mass spectrometry technology and more optimized methods can be applied to the study of clinical samples, and promote the process of individualized treatment. In this study, a quantitative proteomics method based on chemical markers was used to study the protein expression profiles of peripheral blood mononuclear cells (PBMC) in patients with significant efficacy after methotrexate administration, and to combine with clinical data. The biomarkers related to the sensitivity and efficacy of methotrexate are expected to be found to distinguish the sensitive patients and to be used in individual treatment of psoriasis. We selected 20 peripheral blood mononuclear cell samples from 4 patients with effective methotrexate therapy (0 and 8 weeks of treatment) and 4 normal controls. A stable isotope chemochemistry in vitro iTRAQ labeling strategy was used for quantitative proteomics analysis. A total of 3397 proteins were identified, of which 3190 had quantitative information. Through bioinformatics analysis, we found that the expression of histone in patients with good methotrexate treatment changed significantly before and after treatment, and returned to normal level after methotrexate treatment. Our preliminary biological verification of ANXA6 showed that the specific expression of ANXA6 in patients with good methotrexate treatment was high before treatment and returned to normal level after treatment. In this work, we applied drug proteomics to PBMC cells of psoriatic patients for the first time, and found a dynamic protein molecular network related to the efficacy of methotrexate. Our results will provide some guidance for the clinical use of methotrexate and provide new clues for the study of the pharmacological mechanism of methotrexate in the treatment of psoriasis.
【学位授予单位】：中国科学院大学(中国科学院上海药物研究所)
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R96

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