Department of Medical Oncology, Amsterdam UMC Location Vrije Universiteit Amsterdam, Amsterdam, The Netherlands.
Applied Thermal Sciences, Faculty of Science and Technology, University of Twente, Enschede, The Netherlands.
Cancer Med. 2023 May;12(9):10979-10989. doi: 10.1002/cam4.5781. Epub 2023 Mar 14.
In precision oncology, tumor molecular profiles guide selection of therapy. Standardized snap freezing of tissue biospecimens is necessary to ensure reproducible, high-quality samples that preserve tumor biology for adequate molecular profiling. Quenching in liquid nitrogen (LN ) is the golden standard method, but LN has several limitations. We developed a LN -independent snap freezer with adjustable cold sink temperature. To benchmark this device against the golden standard, we compared molecular profiles of biospecimens.
Cancer cell lines and core needle normal tissue biopsies from five patients' liver resection specimens were used to compare mass spectrometry (MS)-based global phosphoproteomic and RNA sequencing profiles and RNA integrity obtained by both freezing methods.
Unsupervised cluster analysis of phosphoproteomic and transcriptomic profiles of snap freezer versus LN -frozen K562 samples and liver biopsies showed no separation based on freezing method (with Pearson's r 0.96 (range 0.92-0.98) and >0.99 for K562 profiles, respectively), while samples with +2 h bench-time formed a separate cluster. RNA integrity was also similar for both snap freezing methods. Molecular profiles of liver biopsies were clearly identified per individual patient regardless of the applied freezing method. Two to 25 s freezing time variations did not induce profiling differences in HCT116 samples.
The novel snap freezer preserves high-quality biospecimen and allows identification of individual patients' molecular profiles, while overcoming important limitations of the use of LN . This snap freezer may provide a useful tool in clinical cancer research and practice, enabling a wider implementation of (multi-)omics analyses for precision oncology.
在精准肿瘤学中,肿瘤分子谱指导治疗选择。为了确保可重复、高质量的样本,以充分进行分子谱分析,有必要对组织生物样本进行标准化的快速冷冻。组织的标准快速冷冻方法是使用液氮(LN)进行急速冷冻,但 LN 存在一些局限性。我们开发了一种具有可调冷阱温度的 LN 独立快速冷冻仪。为了将该设备与黄金标准进行基准比较,我们比较了生物样本的分子谱。
使用来自五名患者肝切除标本的癌细胞系和核心针状正常组织活检,比较两种冷冻方法获得的基于质谱(MS)的全局磷酸化蛋白质组学和 RNA 测序图谱以及 RNA 完整性。
快速冷冻仪与 LN 冷冻 K562 样本和肝活检的磷酸化蛋白质组学和转录组学图谱的无监督聚类分析未显示出基于冷冻方法的分离(Pearson's r 分别为 0.96(范围为 0.92-0.98)和>0.99),而具有+2 h 台时的样本形成了单独的簇。两种快速冷冻方法的 RNA 完整性也相似。无论应用哪种冷冻方法,每个患者的肝活检的分子谱都可以清晰识别。在 HCT116 样本中,2 至 25 秒的冷冻时间变化不会引起分析差异。
新型快速冷冻仪可保存高质量的生物样本,并允许识别个体患者的分子谱,同时克服了 LN 使用的重要限制。该快速冷冻仪可为临床癌症研究和实践提供有用的工具,为精准肿瘤学中的(多)组学分析提供更广泛的实施。