Spatial and molecular resolution of diffuse malignant mesothelioma heterogeneity by integrating label-free FTIR imaging, laser capture microdissection and proteomics.
Spatial and molecular resolution of diffuse malignant mesothelioma heterogeneity by integrating label-free FTIR imaging, laser capture microdissection and proteomics.
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DOI：
10.1038/srep44829
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发表时间：
2017-03-30
影响因子：
4.6
通讯作者：
Gerwert K
中科院分区：
文献类型：
--
作者：
Großerueschkamp F;Bracht T;Diehl HC;Kuepper C;Ahrens M;Kallenbach-Thieltges A;Mosig A;Eisenacher M;Marcus K;Behrens T;Brüning T;Theegarten D;Sitek B;Gerwert K
弥漫性恶性间皮瘤（DMM）是一种异质性恶性肿瘤，表现为三种亚型：上皮样、肉瘤样和双相。DMM表现出高度的空间异质性，这使得对每个亚型中潜在的不同分子过程的彻底理解变得复杂。我们提出了一种新的方法，通过整合FTIR成像和激光捕获显微解剖（LCM），以无标记的方式在空间上解决肿瘤的异质性。随后的蛋白质组分析解剖均质样品提供额外的分子分辨率。FTIR成像以自动和无标签的方式在组织薄切片内解决肿瘤亚型，DMM亚型的准确率约为85%。即使在高度异质的组织结构中，我们的无标记方法也可以识别感兴趣的小区域，可以使用LCM作为均匀样本进行解剖。随后的蛋白质组分析提供了特定位置的分子表征。应用于DMM亚型，我们鉴定了142种差异表达蛋白，包括DMM免疫组织化学面板中常用的5种蛋白质生物标志物。因此，FTIR成像不仅可以解决组织内的形态学改变，还可以解决肿瘤亚型中单个蛋白质水平上的改变。我们的全自动工作流程ftir引导的LCM为从组学研究中收集精确和预测性生物标志物的均匀样品开辟了新的途径。
Diffuse malignant mesothelioma (DMM) is a heterogeneous malignant neoplasia manifesting with three subtypes: epithelioid, sarcomatoid and biphasic. DMM exhibit a high degree of spatial heterogeneity that complicates a thorough understanding of the underlying different molecular processes in each subtype. We present a novel approach to spatially resolve the heterogeneity of a tumour in a label-free manner by integrating FTIR imaging and laser capture microdissection (LCM). Subsequent proteome analysis of the dissected homogenous samples provides in addition molecular resolution. FTIR imaging resolves tumour subtypes within tissue thin-sections in an automated and label-free manner with accuracy of about 85% for DMM subtypes. Even in highly heterogeneous tissue structures, our label-free approach can identify small regions of interest, which can be dissected as homogeneous samples using LCM. Subsequent proteome analysis provides a location specific molecular characterization. Applied to DMM subtypes, we identify 142 differentially expressed proteins, including five protein biomarkers commonly used in DMM immunohistochemistry panels. Thus, FTIR imaging resolves not only morphological alteration within tissue but it resolves even alterations at the level of single proteins in tumour subtypes. Our fully automated workflow FTIR-guided LCM opens new avenues collecting homogeneous samples for precise and predictive biomarkers from omics studies.