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Characterization of cell-free breast cancer patient-derived scaffolds using liquid chromatography-mass spectrometry/mass spectrometry data and RNA sequencing data

机译:使用液相色谱 - 质谱/质谱/质谱数据和RNA测序数据表征无细胞乳腺癌患者衍生的支架

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摘要

Patient-derived scaffolds (PDSs) generated from primary breast cancer tumors can be used to model the tumor microenvironment in vitro. Patient-derived scaffolds are generated by repeated detergent washing, removing all cells. Here, we analyzed the protein composition of 15 decellularized PDSs using liquid chromatography-mass spectrometry/mass spectrometry. One hundred forty-three proteins were detected and their relative abundance was calculated using a reference sample generated from all PDSs. We performed heatmap analysis of all the detected proteins to display their expression patterns across different PDSs together with pathway enrichment analysis to reveal which processes that were connected to PDS protein composition. This protein dataset together with clinical information is useful to investigators studying the microenvironment of breast cancers. Further, after repopulating PDSs with either MCF7 or MDA-MB-231 cells, we quantified their gene expression profiles using RNA sequencing. These data were also compared to cells cultured in conventional 2D conditions, as well as to cells cultured as xenografts in immune-deficient mice. We investigated the overlap of genes regulated between these different culture conditions and performed pathway enrichment analysis of genes regulated by both PDS and xenograft cultures compared to 2D in both cell lines to describe common processes associated with both culture conditions. Apart from our described analyses of these systems, these data are useful when comparing different experimental model systems. Downstream data analyses and interpretations can be found in the research article “Patient-derived scaffolds uncover breast cancer promoting properties of the microenvironment” [1].
机译:从原发性乳腺癌肿瘤产生的患者衍生的支架(PDS)可用于在体外模拟肿瘤微环境。患者衍生的支架由反复洗涤剂洗涤产生,除去所有细胞。在此,使用液相色谱 - 质谱/质谱法分析了15个脱细胞的PDS的蛋白质组成。检测到一百四十三种蛋白质,并且使用从所有PDS产生的参考样品计算它们的相对丰度。我们对所有检测到的蛋白质进行了热映射分析,以将其表达模式与途径富集分析一起显示它们的表达模式,以揭示连接到PD蛋白质组合物的过程。该蛋白质数据集与临床信息一起对研究乳腺癌微环境的研究人员有用。此外,在用MCF7或MDA-MB-231细胞重新挖掘PDS后,我们使用RNA测序量化其基因表达谱。还将这些数据与在常规2D条件下培养的细胞进行比较,以及在免疫缺陷小鼠中培养为异种移植物的细胞。我们研究了这些不同培养条件和通过两种细胞系中的2D调节的基因之间对这些不同培养条件和对异种移植培养物调节的基因的致病分析,以描述与培养条件相关的常用过程。除了我们所描述的这些系统的分析之外,这些数据在比较不同的实验模型系统时很有用。下游数据分析和解释可以在研究制品“患者衍生的支架上发现微环境的乳腺癌促进性能”[1]。

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