MALDI-Mass Spectrometric Imaging Revealing Hypoxia-Driven Lipids and Proteins in a Breast Tumor Model.
MALDI-Mass Spectrometric Imaging Revealing Hypoxia-Driven Lipids and Proteins in a Breast Tumor Model.
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DOI：
10.1021/ac504503x
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发表时间：
2015-06-16
影响因子：
7.4
通讯作者：
Glunde K
中科院分区：
文献类型：
--
作者：
Jiang L;Chughtai K;Purvine SO;Bhujwalla ZM;Raman V;Paša-Tolić L;Heeren RMA;Glunde K
缺氧区域是快速生长的恶性肿瘤及其转移瘤的一个共同特征，并且通常具有空间异质性。缺氧对肿瘤细胞生物学有很大影响，并以多种方式促进肿瘤进展。迄今为止，仅发现了肿瘤缺氧的少数关键分子，例如缺氧诱导因子-1（HIF-1）。生物分子在肿瘤体积中的分布通常是不均匀的，并且可能是由缺氧和 HIF-1α 驱动的。了解肿瘤的空间异质缺氧反应至关重要。质谱成像 (MSI) 提供了一种以高光谱和空间分辨率对组织切片中的生物分子分布进行成像的独特方法。在本文中，使用由 MDA-MB-231-HRE-tdTomato 细胞生长的乳腺肿瘤异种移植物，在 HIF-1α 驱动的含缺氧反应元件 (HRE) 的启动子控制下，具有红色荧光 tdTomato 蛋白构建体，用于检测缺氧区域的空间分布。我们通过对 MDA-MB-231-HRE-tdTomato 乳腺肿瘤异种移植物的 3D 渲染 MSI 体积数据使用主成分分析 - 线性判别分析 (PCA-LDA) 阐明了缺氧区域与脂质和蛋白质定位之间的 3D 空间关系。在这项研究中，我们鉴定了在几个不同途径中活跃的缺氧调节蛋白，例如葡萄糖代谢、肌动蛋白细胞骨架的调节、蛋白质折叠、翻译/核糖体、剪接体、PI3K-Akt信号通路、血红蛋白伴侣、内质网中的蛋白质加工、活性氧的解毒、极光B信号/凋亡执行阶段、RAS信号通路、凋亡中的FAS信号通路/Caspase级联，以及端粒应激诱导衰老。与此同时，我们还确定了缺氧区域和各种脂质种类的共定位，例如 PC(16:0/18:0)、PC(16:0/18:1)、PC(16:0/18:2)、PC(16:1/18:4)、PC(18:0/18:1) 和 PC(18:1/18:1) 等。我们的研究结果揭示了缺氧肿瘤区域的生物分子组成，这可能是特定肿瘤对放射或化疗产生抵抗力的原因。
Hypoxic areas are a common feature of rapidly growing malignant tumors and their metastases and are typically spatially heterogeneous. Hypoxia has a strong impact on tumor cell biology and contributes to tumor progression in multiple ways. To date, only a few molecular key players in tumor hypoxia, such as hypoxia-inducible factor-1 (HIF-1), have been discovered. The distribution of biomolecules is frequently heterogeneous in the tumor volume and may be driven by hypoxia and HIF-1α. Understanding the spatially heterogeneous hypoxic response of tumors is critical. Mass spectrometric imaging (MSI) provides a unique way of imaging biomolecular distributions in tissue sections with high spectral and spatial resolution. In this paper, breast tumor xenografts grown from MDA-MB-231-HRE-tdTomato cells, with a red fluorescent tdTomato protein construct under the control of a hypoxia response element (HRE)-containing promoter driven by HIF-1α, were used to detect the spatial distribution of hypoxic regions. We elucidated the 3D spatial relationship between hypoxic regions and the localization of lipids and proteins by using principal component analysis–linear discriminant analysis (PCA-LDA) on 3D rendered MSI volume data from MDA-MB-231-HRE-tdTomato breast tumor xenografts. In this study, we identified hypoxia-regulated proteins active in several distinct pathways such as glucose metabolism, regulation of actin cytoskeleton, protein folding, translation/ribosome, splicesome, the PI3K-Akt signaling pathway, hemoglobin chaperone, protein processing in endoplasmic reticulum, detoxification of reactive oxygen species, aurora B signaling/apoptotic execution phase, the RAS signaling pathway, the FAS signaling pathway/caspase cascade in apoptosis, and telomere stress induced senescence. In parallel, we also identified colocalization of hypoxic regions and various lipid species such as PC(16:0/18:0), PC(16:0/18:1), PC(16:0/18:2), PC(16:1/18:4), PC(18:0/18:1), and PC(18:1/18:1), among others. Our findings shed light on the biomolecular composition of hypoxic tumor regions, which may be responsible for a given tumor's resistance to radiation or chemotherapy.