An Extensive Metabolomics Workflow to Discover Cardiotoxin-Induced Molecular Perturbations in Microtissues.
An Extensive Metabolomics Workflow to Discover Cardiotoxin-Induced Molecular Perturbations in Microtissues.
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DOI：
10.3390/metabo11090644
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发表时间：
2021-09-21
期刊：
影响因子：
4.1
通讯作者：
Viant MR
中科院分区：
文献类型：
--
作者：
Bowen TJ;Hall AR;Lloyd GR;Weber RJM;Wilson A;Pointon A;Viant MR
发现药物诱导的结构性心脏毒性的作用模式和预测性生物标志物提供了改善先导化合物的心脏安全性评估并在药物开发期间增强临床前到临床转化的潜力。心脏微组织是一个有前途的，生理相关的，在体外模型，每个组成的ca。500个细胞。虽然非靶向代谢组学能够产生关于毒理学作用模式的假设并发现代谢生物标志物，但将该技术应用于悬浮液中的低生物量微组织在实验上具有挑战性。因此，我们首先评估了一种基于过滤的方法收获微组织，并评估了细胞内提取物的纳米电喷雾直接输注质谱（nESI-DIMS）测量的灵敏度和重现性，揭示了由28个合并的微组织组成的样品，通过过滤收获，适用于分析细胞内代谢组和脂质组。随后，实施了一个广泛的工作流程，将细胞内提取物的nESI-DIMS非靶向代谢组学和脂质组学与用过的培养基的超高效液相色谱-质谱（UHPLC-MS/MS）分析相结合，以分析代谢足迹并量化药物暴露浓度。使用合成药物和模型心脏毒素舒尼替尼，时间分辨的代谢和脂质扰动心脏微组织进行了研究，提供了有价值的数据，产生毒理学作用模式的假设和确定推定的生物标志物，如嘌呤代谢和扰动的多不饱和脂肪酸水平的中断。
Discovering modes of action and predictive biomarkers of drug-induced structural cardiotoxicity offers the potential to improve cardiac safety assessment of lead compounds and enhance preclinical to clinical translation during drug development. Cardiac microtissues are a promising, physiologically relevant, in vitro model, each composed of ca. 500 cells. While untargeted metabolomics is capable of generating hypotheses on toxicological modes of action and discovering metabolic biomarkers, applying this technology to low-biomass microtissues in suspension is experimentally challenging. Thus, we first evaluated a filtration-based approach for harvesting microtissues and assessed the sensitivity and reproducibility of nanoelectrospray direct infusion mass spectrometry (nESI-DIMS) measurements of intracellular extracts, revealing samples consisting of 28 pooled microtissues, harvested by filtration, are suitable for profiling the intracellular metabolome and lipidome. Subsequently, an extensive workflow combining nESI-DIMS untargeted metabolomics and lipidomics of intracellular extracts with ultra-high performance liquid chromatography-mass spectrometry (UHPLC-MS/MS) analysis of spent culture medium, to profile the metabolic footprint and quantify drug exposure concentrations, was implemented. Using the synthetic drug and model cardiotoxin sunitinib, time-resolved metabolic and lipid perturbations in cardiac microtissues were investigated, providing valuable data for generating hypotheses on toxicological modes of action and identifying putative biomarkers such as disruption of purine metabolism and perturbation of polyunsaturated fatty acid levels.
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影响因子：
120.1
作者：
Cook, David;Brown, Dearg;Pangalos, Menelas N.
通讯作者：
Pangalos, Menelas N.
DOI：
10.1093/toxsci/kfx097
发表时间：
2017-08-01
期刊：
Toxicological sciences : an official journal of the Society of Toxicology
影响因子：
--
作者：
Brockmeier EK;Hodges G;Hutchinson TH;Butler E;Hecker M;Tollefsen KE;Garcia-Reyero N;Kille P;Becker D;Chipman K;Colbourne J;Collette TW;Cossins A;Cronin M;Graystock P;Gutsell S;Knapen D;Katsiadaki I;Lange A;Marshall S;Owen SF;Perkins EJ;Plaistow S;Schroeder A;Taylor D;Viant M;Ankley G;Falciani F
通讯作者：
Falciani F
人类3D心脏微动物的表征和验证，以评估心脏病理变化。
影响因子：
4.6
作者：
Archer CR;Sargeant R;Basak J;Pilling J;Barnes JR;Pointon A
通讯作者：
Pointon A
影响因子：
3.8
作者：
Doherty, Kimberly R.;Wappel, Robert L.;Bacus, Sarah
通讯作者：
Bacus, Sarah
影响因子：
3.7
作者：
Bordag N;Janakiraman V;Nachtigall J;González Maldonado S;Bethan B;Laine JP;Fux E
通讯作者：
Fux E