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.
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We thank I. Novak and N. Meyn Christensen (Centre for Advanced Bioimaging (CAB), University of Copenhagen) for imaging assistance. We thank J.R. Brewer (University of Southern Denmark) for providing access to his custom-built two-photon microscope. We thank J. Koch for help with decellularization procedures. We thank E. Sahai (Francis Crick Institute) for providing the MATLAB code. We thank J. Couchman, K.B. Jensen (both from Biotech Research & Innovation Centre, University of Copenhagen) and E. Sahai for their critical reading of the manuscript. We thank R. Linding (BRIC, University of Copenhagen) for providing access to mass spectrometry facilities. This work was supported by the Danish Cancer Society (R56-A3342, R124-A7862) (A.E.M.-G. and E.R.H.); the Novo Nordisk Foundation (Hallas Møller Stipend; to C.D.M. and J.T.E.); the European Research Council (ERC-2015-CoG-682881-MATRICAN; to A.E.M.-G. and E.R.H.); the Ragnar Söderberg Foundation Sweden (N19/15; C.D.M.); Cancerfonden Sweden (CAN 2016/283); the Innovation Foundation Denmark (1311-00010B; to T.R.C.); the National Health and Medical Research Council (NHMRC) Australia (APP1129766; T.R.C.); and the Danish Council for Independent Research YDUN grant (1084181001; F.V.A.).
The authors declare no competing financial interests.
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Mayorca-Guiliani, A., Madsen, C., Cox, T. et al. ISDoT: in situ decellularization of tissues for high-resolution imaging and proteomic analysis of native extracellular matrix. Nat Med 23, 890–898 (2017) doi:10.1038/nm.4352
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