Cancer cells facilitate tumor growth by creating favorable tumor micro-environments (TME), altering homeostasis and immune response in the extracellular matrix (ECM) of surrounding tissue. A potential factor that contributes to TME generation and ECM remodeling is the cytoskeleton-associated human death-associated protein kinase 1 (DAPK1). Increased tumor cell motility and de-adhesion (thus, promoting metastasis), as well as upregulated plasminogen-signaling, are shown when functionally analyzing the DAPK1 ko-related proteome. However, the systematic investigation of how tumor cells actively modulate the ECM at the tissue level is experimentally challenging since animal models do not allow direct experimental access while artificial in vitro scaffolds cannot simulate the entire complexity of tissue systems. Here, we used the chorioallantoic membrane (CAM) assay as a natural, collagen-rich tissue model in combination with all-optical experimental access by multiphoton microscopy (MPM) to study the ECM remodeling potential of colorectal tumor cells with and without DAPK1 in situ and even in vivo. This approach demonstrates the suitability of the CAM assay in combination with multiphoton microscopy for studying collagen remodeling during tumor growth. Our results indicate the high ECM remodeling potential of DAPK1 ko tumor cells at the tissue level and support our findings from proteomics.

Clinics of Medicine 1 University Hospital Erlangen Friedrich Alexander University Erlangen Nürnberg 91054 Erlangen Germany

Czech Center for Phenogenomics Institute of Molecular Genetics of the ASCR 142 20 Prague Czech Republic

Department of Pathology and Legal Medicine Federal University of Health Sciences of Porto Alegre Porto Alegre 90050 170 Brazil

Experimental Tumor Pathology Institute of Pathology University Hospital Erlangen Friedrich Alexander University Erlangen Nürnberg 91054 Erlangen Germany

Graduate School in Advanced Optical Technologies Paul Gordan Str 7 91052 Erlangen Germany

Institute of Medical Biotechnology Friedrich Alexander University Erlangen Nürnberg Paul Gordan Str 3 91052 Erlangen Germany

Institute of Pathology University Hospital Erlangen Friedrich Alexander University Erlangen Nürnberg 91054 Erlangen Germany

School of Mechanical Medical and Process Engineering Queensland University of Technology Brisbane QLD 4000 Australia

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