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Direct investigation of the early cellular changes induced by metastatic cells within the surrounding tissue remains a challenge. Here we present a system in which metastatic cancer cells release a cell-penetrating fluorescent protein, which is taken up by neighbouring cells and enables spatial identification of the local metastatic cellular environment. Using this system, tissue cells with low representation in the metastatic niche can be identified and characterized within the bulk tissue. To highlight its potential, we applied this strategy to study the cellular environment of metastatic breast cancer cells in the lung. We report the presence of cancer-associated parenchymal cells, which exhibit stem-cell-like features, expression of lung progenitor markers, multi-lineage differentiation potential and self-renewal activity. In ex vivo assays, lung epithelial cells acquire a cancer-associated parenchymal-cell-like phenotype when co-cultured with cancer cells and support their growth. These results highlight the potential of this method as a platform for new discoveries. A cell-penetrating fluorescent marker is used to label cells in the metastatic tumour microenvironment, revealing a variety of cell types including parenchymal cells with lung stem-cell characteristics., Author(s): Luigi Ombrato [sup.1] , Emma Nolan [sup.1] , Ivana Kurelac [sup.1] [sup.2] , Antranik Mavousian [sup.3] , Victoria Louise Bridgeman [sup.1] , Ivonne Heinze [sup.4] , Probir Chakravarty [sup.5] [...]

An Amendment to this paper has been published and can be accessed via a link at the top of the paper., Author(s): Luigi Ombrato [sup.1] , Emma Nolan [sup.1] , Ivana Kurelac [sup.1] [sup.2] , Antranik Mavousian [sup.3] , Victoria Louise Bridgeman [sup.1] , Ivonne Heinze [sup.4] , Probir Chakravarty [sup.5] [...]