H&TRC - Health & Technology Research Center, Escola Superior de Tecnologia da Saúde de Lisboa (ESTeSL), Instituto Politécnico de Lisboa, Avenida D. João II, lote 4.69.01, Parque das Nações, 1990-096 Lisboa, Portugal; NOVA National School of Public Health, Public Health Research Centre, Universidade NOVA de Lisboa, Lisbon, Portugal; Comprehensive Health Research Center (CHRC), Universidade NOVA de Lisboa, Portugal.
Comprehensive Health Research Center (CHRC), Universidade NOVA de Lisboa, Portugal; ISEL - Instituto Superior de Engenharia de Lisboa, Instituto Politécnico de Lisboa, R. Conselheiro Emidio Navarro 1, 1959‑007 Lisboa, Portugal; NOVA Medical School, Faculdade de Ciências Médicas, Universidade NOVA de Lisboa, 1169-056 Lisbon, Portugal.
Mutat Res Genet Toxicol Environ Mutagen. 2023 Oct;891:503681. doi: 10.1016/j.mrgentox.2023.503681. Epub 2023 Aug 18.
Genotoxicity is an important information that should be included in human biomonitoring programmes. However, the usually applied cytogenetic assays are laborious and time-consuming, reason why it is critical to develop rapid and economic new methods. The aim of this study was to evaluate if the molecular profile of frozen whole blood, acquired by Fourier Transform Infrared (FTIR) spectroscopy, allows to assess genotoxicity in occupational exposure to antineoplastic drugs, as obtained by the cytokinesis-block micronucleus assay. For that purpose, 92 samples of peripheral blood were studied: 46 samples from hospital professionals occupationally exposed to antineoplastic drugs and 46 samples from workers in academia without exposure (controls). It was first evaluated the metabolome from frozen whole blood by methanol precipitation of macromolecules as haemoglobin, followed by centrifugation. The metabolome molecular profile resulted in 3 ratios of spectral bands, significantly different between the exposed and non-exposed group (p < 0.01) and a spectral principal component-linear discriminant analysis (PCA-LDA) model enabling to predict genotoxicity from exposure with 73 % accuracy. After optimization of the dilution degree and solution used, it was possible to obtain a higher number of significant ratios of spectral bands, i.e., 10 ratios significantly different (p < 0.001), highlighting the high sensitivity and specificity of the method. Indeed, the PCA-LDA model, based on the molecular profile of whole blood, enabled to predict genotoxicity from the exposure with an accuracy, sensitivity, and specificity of 92 %, 93 % and 91 %, respectively. All these parameters were achieved based on 1 μL of frozen whole blood, in a high-throughput mode, i.e., based on the simultaneous analysis of 92 samples, in a simple and economic mode. In summary, it can be conclude that this method presents a very promising potential for high-dimension screening of exposure to genotoxic substances.
遗传毒性是人体生物监测计划中应包含的重要信息。然而，通常应用的细胞遗传学检测方法既繁琐又耗时，因此开发快速、经济的新方法至关重要。本研究旨在评估傅里叶变换红外（FTIR）光谱法获取的冷冻全血分子谱是否可用于评估职业性接触抗肿瘤药物的遗传毒性，方法为细胞有丝分裂阻断微核试验。为此，研究了 92 份外周血样本：46 份来自医院职业性接触抗肿瘤药物的专业人员，46 份来自学术界无接触的工人（对照组）。首先通过甲醇沉淀大分子（如血红蛋白）沉淀冷冻全血的代谢组，然后进行离心。代谢组分子谱得到 3 个光谱带比值，在暴露组和非暴露组之间存在显著差异（p<0.01），并通过光谱主成分-线性判别分析（PCA-LDA）模型预测暴露的遗传毒性，准确率为 73%。优化稀释度和溶液后，可获得更多的光谱带比值显著差异（p<0.001），从而提高方法的灵敏度和特异性。事实上，基于全血分子谱的 PCA-LDA 模型可实现 92%的准确性、93%的灵敏度和 91%的特异性，从暴露预测遗传毒性。所有这些参数均基于 1μL 冷冻全血，以高通量模式进行，即基于同时分析 92 个样本，以简单经济的模式进行。总之，可以得出结论，该方法在高通量筛选遗传毒性物质暴露方面具有很大的应用潜力。
