A DNA-nanoassembly-based approach to map membrane protein nanoenvironments

Cell-to-cell communication pathways coordinate cellular functions in multicellular organisms and are frequently dysregulated in cancer, allowing cells to acquire abnormal functions such as invasion of neighboring tissues. Membrane proteins are key sensor components in cell-to-cell communication and form the largest class of proteins in the druggable genome. We hypothesize that the spatial distribution of ligands and receptors contributes to defining the fundamental mechanisms of action of cell-to-cell communication pathways and their cellular outcomes. The research program relies on the development of tools that provide well-defined physical inputs to cells, not confounded by simultaneous changes in chemical inputs. Therefore, beyond state-of-the-art developments in nanotechnology are here integrated with cell biology. In particular, DNA origami technology is applied to the development of ligand nanoclusters with customized spatial organization and mechanical properties. We will focus on the ephrin/Eph and DSL/Notch cell-to-cell communication pathways, which regulate embryonic development and the homeostasis of adult organs. These pathways are commonly disrupted in cancer, showing tumour suppressing or tumour promoting character. The mechanisms that generate the diversity of outcomes in normal and disease settings are largely unknown. We will use DNA origami/ephrin ligand nanoclusters to investigate whether the spatial organization ligand assemblies impact receptor function. We aim to generate a knowledge foundation for the development of new rational therapeutic approaches that target ephrin ligand/Eph and Notch receptor signalling.