Dr. Margarete Fischer-Bosch-Institute of Clinical Pharmacology , Stuttgart , Germany and University of Tübingen, Tübingen, Germany.
Department of Clinical Pharmacology , University Hospital Tübingen , Tübingen , Germany.
J Proteome Res. 2018 Sep 7;17(9):3039-3049. doi: 10.1021/acs.jproteome.8b00199. Epub 2018 Aug 23.
Tissue analysis represents a powerful tool for the investigation of disease pathophysiology. However, the heterogeneous nature of tissue samples, in particular of neoplastic, may affect the outcome of such analysis and hence obscure interpretation of results. Thus, comprehensive isolation and extraction of transcripts and metabolites from an identical tissue specimen would minimize variations and enable the economic use of biopsy material which is usually available in limited amounts. Here we demonstrate a fast and simple protocol for combined transcriptomics and metabolomics analysis in homogenates prepared from one single tissue sample. Metabolites were recovered by protein precipitation from lysates originally prepared for RNA isolation and were analyzed by LC-QTOF-MS after HILIC and RPLC separation, respectively. Strikingly, although ion suppression was observed, over 80% of the 2885 detected metabolic features could be extracted and analyzed with high reproducibility (CV ≤ 20%). Moreover fold changes of different tumor and nontumor kidney tissues were correlated to an established metabolomics protocol and revealed a strong correlation ( r ≥ 0.75). In order to demonstrate the feasibility of the combined analysis of RNA and metabolites, the protocol was applied to kidney tissue of metformin treated mice to investigate drug induced alterations.
组织分析是研究疾病病理生理学的有力工具。然而，组织样本的异质性，特别是肿瘤组织的异质性，可能会影响此类分析的结果，从而使结果的解释变得模糊。因此，从相同的组织标本中综合分离和提取转录本和代谢物，可以最大限度地减少变异，并使活检材料得到经济利用，而活检材料通常数量有限。在这里，我们展示了一种从单个组织样本的匀浆中进行转录组学和代谢组学联合分析的快速而简单的方案。代谢物通过从最初用于 RNA 分离的裂解物中沉淀蛋白质来回收，并在分别进行亲水相互作用色谱（HILIC）和反相高效液相色谱（RPLC）分离后，通过 LC-QTOF-MS 进行分析。值得注意的是，尽管观察到离子抑制，但可以提取和分析超过 80%的 2885 个检测到的代谢特征，具有很高的重现性（CV ≤ 20%）。此外，不同肿瘤和非肿瘤肾脏组织的倍数变化与已建立的代谢组学方案相关，相关性很强（r≥0.75）。为了证明 RNA 和代谢物联合分析的可行性，该方案应用于二甲双胍处理的小鼠肾脏组织，以研究药物诱导的变化。
