PO.MCB09.04 · 分子与细胞生物学
CSF代谢组学揭示甲氨蝶呤治疗的幼年大鼠模型中一碳代谢及磷脂酰胆碱丰度的改变
CSF metabolomics reveals alterations in one-carbon metabolism and phosphatidylcholine abundance in a methotrexate-treated juvenile rat model
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摘要 Abstract
中文摘要
背景：儿童急性淋巴细胞白血病(ALL)的治疗虽通常可治愈，但与神经毒性相关，并导致40-70%的幸存者出现化疗相关认知障碍(CRCI)，显著影响其生活质量。甲氨蝶呤(MTX)是ALL化疗方案的关键组成部分，是CRCI的主要促成因素。利用脑脊液(CSF)代谢组学，我们既往工作显示接受化疗的儿童患者表现出脂质代谢改变，尤其是磷脂酰胆碱(PC)。然而，由于ALL化疗涉及多种药物，MTX对这些代谢改变的具体贡献仍不清楚。
实验方法：使用旨在于儿科相关背景下分离MTX特异性效应的幼年大鼠模型，我们在3至8周龄之间给予六次腹腔内(每剂0.5 mg/kg)和四次鞘内(每剂1 mg/kg)MTX注射。为表征MTX诱导的代谢变化，我们在首次腹腔内注射时和第四次注射时进行CSF代谢组学分析。在末次注射后五周，我们分别使用物体位置(OP)和新物体识别(OR)行为测试，评估MTX暴露是否相对于PBS处理对照损害空间和视觉记忆。
结果：与对照相比，MTX处理的大鼠表现出空间和视觉记忆损害。与我们既往结果及MTX作用机制一致，MTX处理动物CSF中一碳代谢的决定因素随时间下调。这包括S-腺苷甲硫氨酸(SAM，FC = 0.36，p-adj = 0.02)和甲硫氨酸(FC = 3.24E-06，p-adj = 6.41E-06)。相反，转硫途径代谢物如胱硫醚(FC = 3.98，p-adj = 0.001)和半胱氨酸(FC = 2.81，p-adj = 0.02)上调。最后，与对照相比，MTX治疗还诱导了脂质代谢改变，8种缩醛磷脂和9种PC显著过表达，与我们在接受化疗的儿童ALL患者CSF中的发现一致。
结论：尽管MTX已使用逾八十年，但其作用机制和副作用均未被完全阐明。因此，临床前模型对于界定MTX对化疗相关神经毒性的贡献仍具重要意义。这项工作连同我们近期在人体中的发现表明，CSF代谢组学可通过预测性生物标志物实现对CRCI高危患者的早期识别，并指导未来的神经保护干预。
查看英文原文 English abstract
Background:Although typically curative, treatment for pediatric acute lymphoblastic leukemia (ALL) is associated with neurotoxicity and leads to chemotherapy-related cognitive impairment (CRCI) in 40-70% of survivors, significantly impacting their quality of life. Methotrexate (MTX), a key component of ALL chemotherapy regimens, is a major contributor to CRCI. Using cerebrospinal fluid (CSF) metabolomics, our previous work showed that pediatric patients undergoing chemotherapy exhibited alterations in lipid metabolism, particularly phosphatidylcholines (PC). However, because ALL chemotherapy involves multiple agents, the specific contribution of MTX to these metabolic alterations remains unclear.
Experimental procedures:Using a juvenile rat model designed to isolate MTX-specific effects within a pediatric-relevant context, we administered six intraperitoneal (0.5 mg/kg per dose) and four intrathecal (1 mg/kg per dose) MTX injections between 3 and 8 weeks of age. To characterize MTX-induced metabolic changes, we performed CSF metabolomics at the time of the first intraperitoneal injection and at the fourth. Five weeks after the last injection, we assessed whether MTX exposure impaired spatial and visual memory using Object Placement (OP) and novel Object Recognition (OR) behavioral tests, respectively, in comparison to PBS-treated controls.
Results:MTX-treated rats exhibited spatial and visual memory impairments compared with controls. In accordance with our previous results and MTX's mechanism of action, determinants of one-carbon metabolism were downregulated in the CSF of MTX-treated animals over time. This includes S-adenosylmethionine (SAM, FC = 0.36, p-adj = 0.02) and methionine (FC = 3.24E-06, p-adj = 6.41E-06). In contrast, transsulfuration pathway metabolites such as cystathionine (FC = 3.98, p-adj = 0.001) and cysteine (FC = 2.81, p-adj = 0.02) were upregulated. Last, MTX treatment also induced alterations in lipid metabolism compared to controls, with a significant over-representation of 8 plasmalogens and 9 PC, consistent with our findings in the CSF of pediatric ALL patients undergoing chemotherapy.
Conclusion:Although in use for more than eight decades, neither the mechanism of action nor the side effects of MTX are fully understood. Preclinical models therefore remain instrumental for defining MTX's contribution to chemotherapy-related neurotoxicity. This work, together with our recent findings in humans, demonstrates that CSF metabolomics may enable the early identification of patients at risk for CRCI through predictive biomarkers and guide future neuroprotective interventions.
利益披露 Disclosure
J. Willekens, None..
C. Patel, None.