Dekker Lennard J M, Verheul Cassandra, Wensveen Nicky, Leenders William, Lamfers Martine L M, Leenstra Sieger, Luider Theo M
Laboratories of Neuro-Oncology/Clinical and Cancer Proteomics, Department of Neurology, Erasmus MC, Rotterdam, P.O. Box 2040, 3000 CA Rotterdam, The Netherlands.
Department of Neurosurgery, Erasmus MC, Rotterdam, P.O. Box 2040, 3000 CA Rotterdam, The Netherlands.
ACS Omega. 2022 Jan 19;7(4):3568-3578. doi: 10.1021/acsomega.1c06121. eCollection 2022 Feb 1.
The R132H mutation in the metabolic enzyme isocitrate dehydrogenase 1 (IDH1) is the most important prognostic factor for the survival of glioma patients. Subsequent studies led to the discovery of a panel of enzymes mainly involved in glutamate anaplerosis and aerobic glycolysis that change in abundance as a result of the mutation. To further study these changes, appropriate glioma models are required that accurately mimic metabolism. To investigate how metabolism is affected by cell culture, we here compared surgically obtained snap-frozen glioma tissues with their corresponding primary glioma cell culture models with a previously developed targeted mass spectrometry proteomic assay. We determined the relative abundance of a panel of metabolic enzymes. Results confirmed increased glutamate use and decreased aerobic glycolysis in resected IDH1 R132H glioma tissue samples. However, these metabolic profiles were not reflected in the paired glioma primary cell cultures. We suggest that culture conditions and tumor microenvironment play a crucial role in maintaining the metabolic situation in cell culture models. For this reason, new models that more closely resemble the microenvironment, such as three-dimensional cell co-cultures or organotypic multicellular spheroid models, need to be developed and investigated.
代谢酶异柠檬酸脱氢酶1（IDH1）中的R132H突变是胶质瘤患者生存的最重要预后因素。随后的研究发现了一组主要参与谷氨酸回补和有氧糖酵解的酶，它们的丰度因该突变而发生变化。为了进一步研究这些变化，需要合适的胶质瘤模型来准确模拟代谢。为了研究细胞培养如何影响代谢，我们在此使用先前开发的靶向质谱蛋白质组学分析方法，将手术获取的速冻胶质瘤组织与其相应的原发性胶质瘤细胞培养模型进行了比较。我们测定了一组代谢酶的相对丰度。结果证实，在切除的IDH1 R132H胶质瘤组织样本中，谷氨酸的利用增加，有氧糖酵解减少。然而，这些代谢特征在配对的胶质瘤原代细胞培养中并未体现。我们认为培养条件和肿瘤微环境在维持细胞培养模型中的代谢状况方面起着关键作用。因此，需要开发和研究更接近微环境的新模型，如三维细胞共培养或器官型多细胞球体模型。
