Thomas R. Cox, Jun 2017
“We’re seeing things we’ve never seen before”: groundbreaking new technique sheds light on the ‘matrix’ surrounding our cells
Our most recent research has just been published in Nature Medicine.
In our paper we describe a new and intuitive new way to dissolve cells from tissues, leaving behind the extracellular matrix (ECM) or ‘matrix’.
The matrix is made up of 100’s of differing building blocks and surrounds the cells in our body. It is incredibly important in the progression and spread of cancer – but up until now it has been notoriously difficult to study in detail.
What did we do?
The work, which began in 2013, was conceived and carried out at the Biotech Research and Innovation Centre (BRIC), part of the University of Copenhagen by a small team of dynamic young PostDocs – Alejandro Mayorca-Guiliani, Chris D. Madsen and Thomas R. Cox – and was led by Professor Janine Erler.
We wanted to find a better way to see and study the matrix in cancer. To do this we developed a novel approach to dissolve cells from tissues, leaving behind the matrix which would allow us to study it in exquisite detail.
For years the 3-dimensional structure of the matrix has been difficult to study and previous approaches to enhance this have been limited. However, by using the blood vessels in organs and tissues we can deliver reagents that melt away the cells but leave the matrix in place. Now, when we look at the organs under the microscope we can see a whole new world of detail in 3D.
Not only are we seeing that the matrix is very different between different tumour types, but when a tumour spreads around the body (metastasizes), the matrix is also drastically different between the primary tumour and the metastases. This is true in terms of both the specific composition of the building blocks that make up the matrix, but also in the way that they are put together. This is telling us that remodelling of the matrix in cancer is organ-specific, and that has important implications for future therapy.
The road ahead
Moving forward we are now working to understand what all these changes mean. Most of what we see has not been observed before so now the task is to understand how and why they might be important in the progression and spread of cancer.
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 mouse tissues and disruption of these patterns in diseased tissues. The ISDoT procedure allows for the study of native ECM structure under normal and pathological conditions in unprecedented detail.
View the article on the Nature Medicine website
Mayorca-Guiliani AE*, Madsen CD*, Cox TR*, Horton ER, Venning FA, Erler JT
ISDoT: In Situ Decellularisation of Tissues for high resolution imaging and proteomic analysis of native extracellular matrix
Nature Medicine 12 Jun 2017
* Co-first authors
This work was supported by the Danish Cancer Society; the Novo Nordisk Foundation (Hallas Møller Stipend) the European Research Council; the Ragnar Söderberg Foundation Sweden; the Innovation Foundation Denmark; the National Health and Medical Research Council (NHMRC) Australia; and the Danish Council for Independent Research YDUN grant.