肾移植受者嘌呤类药物不良反应缘由：NDPK/TGTP/Rac1代谢通路关键基因变异相关研究

嘌呤类药物临床应用广泛，但其不良反应发生率高是面临的最大挑战。核苷二磷酸激酶（NDPK）在体内催化产生嘌呤类药物活性代谢物硫鸟嘌呤核苷三磷酸（TGTP），而TGTP直接作用于分子靶标小G蛋白Rac1发挥作用，因此NDPK/TGTP/Rac1代谢通路在该类药物反应中起关键作用。本课题采用"两阶段分析"关联研究策略，以服用嘌呤类药物硫唑嘌呤的肾移植受者为主要研究对象，首先检测分析红细胞中NDPK和Rac1活性、蛋白含量，TGTP浓度，药物不良反应间相关性。其次，对编码NDPK和Rac1的关键基因NME和RAC1遗传变异与不良反应关联性进行全面分析，重点探讨基因-基因交互作用的影响。同时结合分子和细胞生物学技术手段，深入探讨相关联基因变异的生物学功能。本研究将系统阐明NDPK/TGTP/Rac1代谢通路关键基因变异在嘌呤类药物不良反应中的作用机制，可为提高其临床应用安全性提供新思路和实验基础。

Thiopurines have been widely used in a variety of clinical conditions, however, their clinical use is frequently associated with a relatively high incidence of adverse reactions. Nucleoside diphosphate kinase (NDPK; EC 2.7.4.6) can catalyze the conversion of 6-thioguanosine diphosphate (TGDP) into the active metabolite 6-thioguanosine triphosphate (TGTP). Furthermore, the molecular mechanism of immunosuppression by thiopurines is due to the specific end-metabolite 6-TGTP that binds and inhibits the function of the small guanosine triphosphatase (GTPase) Rac1 in activated T-lymphocytes leading to apoptosis. Therefore, NDPK/TGTP/Rac1 metabolite pathway plays a pivotal role in the drug response of thiopurines. In the present study, a "two-stage analysis" method is employed to investigate the association. Firstly, we will determine and analyze the activity and protein expression of NDPK and Rac1 in human erythrocytes, the TGTP concentration, as well as their relationships with thiopurine-induced adverse drug reactions in Chinese kidney transplant recipients. Secondly, we will carry out a comprehensive investigation to ascertain the association between genetic variations in their encoded NME and RAC1 genes and the thiopurine-induced adverse drug reactions, particularly the influence of gene-gene interplays. Furthermore, we will investigate the underlying biological functions of genetic variations associated with adverse drug reactions via molecular biology, cell biology techniques. Thus, a deep insight into genetic variations in key genes in NDPK/TGTP/Rac1 metabolic pathway and their underlying mechanisms in thiopurine-induced adverse drug reactions in Chinese kidney transplant recipients will be illuminated, which can provide new ideas and experimental foundations for improving drug safety of thiopurines therapy.