Hurkmans Evelien G E, Klumpers Marije J, Vermeulen Sita H, Hagleitner Melanie M, Flucke Uta, Schreuder H W Bart, Gelderblom Hans, Bras Johannes, Guchelaar Henk-Jan, Coenen Marieke J H, Te Loo D Maroeska W M
Department of Human Genetics, Radboud Institute for Health Sciences, Radboud University Medical Center, Nijmegen, Netherlands.
Department of Pediatrics, Radboud Institute for Molecular Life Sciences, Radboud University Medical Center, Nijmegen, Netherlands.
Front Pharmacol. 2020 Aug 12;11:1241. doi: 10.3389/fphar.2020.01241. eCollection 2020.
High-dose methotrexate is a cornerstone agent in the chemotherapeutic treatment of patients with osteosarcoma. However, patients often develop methotrexate-induced toxicities. We aim to identify determinants of methotrexate-induced toxicities in osteosarcoma patients by investigating the relation between drug plasma levels, methotrexate-induced toxicities, and germline variants in genes related to drug absorption, distribution, metabolism, and elimination. A cohort of 114 osteosarcoma patients was genotyped for 1,931 variants in 231 genes using the Drug Metabolism Enzymes and Transporters Plus array. Methotrexate plasma levels and laboratory measurements during and after high-dose methotrexate treatment concerning renal function, liver damage, and myelopoiesis to reflect toxicity outcomes were obtained. One hundred and thirteen patients and a subset of 545 variants in 176 genes passed quality control checks. Methotrexate plasma levels showed associations with creatinine, alanine aminotransferase, and hemoglobin. Genetic variant rs3736599 in the 5'-untranslated region of was associated with lower 48 hour methotrexate plasma levels [coef -0.313 (95% CI -0.459 - -0.167); p = 2.60 × 10]. Association with methotrexate-induced decreased thrombocyte counts was found for two intronic variants in {rs4803418 [coef -0.187 (95% CI -0.275 - -0.099); p = 3.04 × 10] and rs4803419 [coef -0.186 (95% CI -0.278 - -0.093); p = 8.80 × 10]}. An association with increased thrombocyte counts was identified for the intronic variant rs4808326 in [coef 0.193 (95% CI 0.099 - 0.287); p = 6.02 × 10]. Moreover, a secondary analysis with a binary approach using CTCAE toxicity criteria resulted in a nominal significant associations (p < 0.05) for two out of three variants (rs4803418 and rs4808326). This is the first study to identify genetic variants in , and to be associated with methotrexate pharmacokinetics and toxicities. Validation of these variants in an independent cohort and further functional investigation of variants in the identified genes is needed to determine if and how they affect methotrexate plasma levels and the development of methotrexate-induced toxicities.
大剂量甲氨蝶呤是骨肉瘤患者化疗治疗的基石药物。然而，患者常出现甲氨蝶呤诱导的毒性反应。我们旨在通过研究药物血浆水平、甲氨蝶呤诱导的毒性反应以及与药物吸收、分布、代谢和排泄相关基因的种系变异之间的关系，确定骨肉瘤患者甲氨蝶呤诱导毒性反应的决定因素。使用药物代谢酶和转运体加强版芯片对114例骨肉瘤患者的231个基因中的1931个变异进行基因分型。获取大剂量甲氨蝶呤治疗期间及之后反映毒性结果的甲氨蝶呤血浆水平以及有关肾功能、肝损伤和骨髓生成的实验室测量值。113例患者以及176个基因中的545个变异子集通过了质量控制检查。甲氨蝶呤血浆水平与肌酐、丙氨酸转氨酶和血红蛋白存在关联。基因rs3736599位于的5'-非翻译区，与48小时甲氨蝶呤血浆水平较低相关[系数-0.313（95%可信区间-0.459至-0.167）；p = 2.60×10]。发现位于中的两个内含子变异与甲氨蝶呤诱导的血小板计数减少相关{rs4803418[系数-0.187（95%可信区间-0.275至-0.099）；p = 3.04×10]和rs4803419[系数-0.186（95%可信区间-0.278至-0.093）；p = 8.80×10]}。确定位于中的内含子变异rs4808326与血小板计数增加相关[系数0.193（95%可信区间0.099至0.287）；p = 6.02×10]。此外，使用CTCAE毒性标准的二元方法进行的二次分析显示，三个变异中的两个（rs4803418和rs4808326）存在名义上的显著关联（p < 0.05）。这是第一项确定、和中的基因变异与甲氨蝶呤药代动力学及毒性相关的研究。需要在独立队列中验证这些变异，并对已鉴定基因中的变异进行进一步功能研究，以确定它们是否以及如何影响甲氨蝶呤血浆水平和甲氨蝶呤诱导毒性反应的发生。
