Biotech Research and Innovation Centre (BRIC), University of Copenhagen (UCPH), Copenhagen, Denmark.
Department of Laboratory Medicine, Division of Translational Cancer Research, Lund University, Lund, Sweden.
Nat Med. 2017 Jul;23(7):890-898. doi: 10.1038/nm.4352. Epub 2017 Jun 12.
The extracellular matrix (ECM) is a master regulator of cellular phenotype and behavior. It has a crucial role in both normal tissue homeostasis and disease pathology. Here we present a fast and efficient approach to enhance the study of ECM composition and structure. Termed in situ decellularization of tissues (ISDoT), it allows whole organs to be decellularized, leaving native ECM architecture intact. These three-dimensional decellularized tissues can be studied using high-resolution fluorescence and second harmonic imaging, and can be used for quantitative proteomic interrogation of the ECM. Our method is superior to other methods tested in its ability to preserve the structural integrity of the ECM, facilitate high-resolution imaging and quantitatively detect ECM proteins. In particular, we performed high-resolution sub-micron imaging of matrix topography in normal tissue and over the course of primary tumor development and progression to metastasis in mice, providing the first detailed imaging of the metastatic niche. These data show that cancer-driven ECM remodeling is organ specific, and that it is accompanied by comprehensive changes in ECM composition and topological structure. We also describe differing patterns of basement-membrane organization surrounding different types of blood vessels in healthy and diseased tissues. The ISDoT procedure allows for the study of native ECM structure under normal and pathological conditions in unprecedented detail.
细胞外基质 (ECM) 是细胞表型和行为的主要调节剂。它在正常组织稳态和疾病病理学中都起着至关重要的作用。在这里，我们提出了一种快速有效的方法来增强 ECM 组成和结构的研究。该方法称为组织原位脱细胞化 (ISDoT)，可使整个器官脱细胞化，保留天然 ECM 结构的完整性。这些三维脱细胞组织可用于高分辨率荧光和二次谐波成像研究，并可用于 ECM 的定量蛋白质组学研究。与其他测试方法相比，我们的方法在保留 ECM 结构完整性、促进高分辨率成像和定量检测 ECM 蛋白方面具有优势。特别是，我们对正常组织和小鼠原发性肿瘤发展和转移过程中的 ECM 拓扑结构进行了亚微米级的高分辨率成像，首次详细描绘了转移龛的图像。这些数据表明，癌症驱动的 ECM 重塑是器官特异性的，并且伴随着 ECM 组成和拓扑结构的全面变化。我们还描述了健康和患病组织中不同类型血管周围基底膜组织的不同模式。ISDoT 程序允许在前所未有的细节水平下研究正常和病理条件下的天然 ECM 结构。
