Establishment of personalized immunosuppressive therapy based on molecular mechanisms of transplant immunological network

1.人体组织标本的基因表达谱：检测了MDR1、CYP3A4、CYP3A5、CYP3A7和CYP3A43在天然肠（n=192）和移植物肝（n=208）部分组织标本中的mRNA表达水平和遗传多态性。手术时天然小肠中MDR1的mRNA表达水平是活体肝移植后立即调整他克莫司初始剂量的重要生物标志物。移植肝中CYP3A5的单核苷酸多态性（SNP）数据与术后他克莫司剂量的增加有显著相关。澄清与免疫抑制剂个体差异相关的药代动力学和药效学因素：群体药代动力学分析发现，手术时MDR1 mRNA表达水平、CYP3A5*3 SNP、移植物与受体体重比以及经典临床试验标志物是具有统计学意义的固定效应。此外，我们发现，基于对手术时肠道MDR1表达水平的评估，他克莫司初始给药方案成功地将移植后急性细胞排斥反应的频率降低到30%左右。外周血白细胞基因表达信息的临床意义：针对活体肝移植术后急性细胞排斥反应的发生，采用术后3、7、14天外周血总RNA检测基因表达水平。在500点以上的临床汇总样本中，他克莫司的靶治疗水平倾向于血细胞中MDR1 mRNA高表达的患者，而不是糖皮质激素受体（GR/NR3C1）高表达的患者。这些结果提示，在活体肝移植患者体内，多种组织蛋白如天然小肠和外周白细胞的MDR1 mRNA水平、移植物肝脏和天然肠道的CYP3A5基因型等是影响其药代动力学和药效学的因素。通过整合这些分子因素，必须建立预防肝移植后急性细胞排斥反应的策略。然而，个体间对他克莫司反应差异的分子机制有待进一步研究。少

1.Gene expression profile using human tissue specimens : The mRNA expression levels and genetic polymorphisms of MDR1,CYP3A4,CYP3A5,CYP3A7 and CYP3A43 were examined in the part of tissue specimens of native intestine (n=192) and graft liver (n=208). The mRNA expression level of MDR1 in the native small intestine at surgery was revealed to be a significant biomarker for adjustment of initial dosage of tacrolimus immediately after the living-donor liver transplantation. The data of single nucleotide polymorphism (SNP) of CYP3A5 in the graft liver was significantly associated with the dose escalation of tacrolimus by postoperative days.2.Clarification of the pharmacokinetic- and pharmacodymanic-factors to associate the individual variation of immunosuppressant : The mRNA expression level of MDR1 at surgery, CYP3A5*3 SNP and graft-vs-recipients body weight ratio as well as the classical clinical test markers were found to be the statistical significant fixed effects by the population pharm … More acokinetic analyses. In addition, we found that the frequency of posttransplant acute cellular rejection was successfully reduced to around 30% by the initial dosage regimen tacrolimus based on the evaluation of intestinal expression level of MDR1 at surgery.3.Clinical significance of gene expression information in the peripheral blood leukocyte : Focusing on the occurrence of acute cellular rejection after living-donor liver transplantation, the gene expression levels were examined with the total RNA fractions from the peripheral blood cells at postoperative days 3, 7 and 14. Using the pooled clinical samples over 500 points, the target therapeutic level of tacrolimus tended to be higher in the patients with high-expression level of MDR1 mRNA in blood cells rather than glucocorticoid receptor (GR/NR3C1).These results suggested that various proteins in the tissues such as the MDR1 mRNA level in the native small intestine and peripheral leukocytes and CYP3A5 genotype in the graft liver and native intestine were pharmacokinetic and pharmacodynamic factors in living-donor liver transplant patients. By integrating these molecular factors, the strategy to prevent acute cellular rejection after liver transplantation must be established. However, the molecular mechanisms of interindividual variation of the response against tacrolimus should be clarified in future. Less