Novel microscopy-based screening method reveals regulators of contact-dependent intercellular transfer

• DOI: 10.1038/srep12879
• Publication Date: 2015-08-14T09:14:19Z
• Authors: Dominik Michael Frei; Erlend Hodneland; Ivan Rios-Mondragon; Anne Burtey; Beate Neumann; Jutta Bulkescher; Julia Schölermann; Rainer Pepperkok; Hans-Hermann Gerdes; Tanja Kögel
• Keywords: 570; [SDV.BIO]Life Sciences [q-bio]/Biotechnology; [SDV.IB.IMA]Life Sciences [q-bio]/Bioengineering/Imaging; [SDV.BBM]Life Sciences [q-bio]/Biochemistry; [SDV.SA.AGRO]Life Sciences [q-bio]/Agricultural sciences/Agronomy; 610; [SDV.CAN]Life Sciences [q-bio]/Cancer; [SDV.BC]Life Sciences [q-bio]/Cellular Biology; Cell Communication; Myosins; Article; GTP Phosphohydrolases; [SDV.EE.ECO]Life Sciences [q-bio]/Ecology, environment/Ecosystems; environment/Health; Lab-On-A-Chip Devices; environment/Symbiosis; Humans; [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology; [SDV.IB.BIO]Life Sciences [q-bio]/Bioengineering/Biomaterials; Molecular Biology; [SDV.EE.SANT]Life Sciences [q-bio]/Ecology, environment/Health; [SDV.BA.MVSA]Life Sciences [q-bio]/Animal biology/Veterinary medicine and animal Health; Microscopy, Confocal; Cell Membrane; Molecular Imaging; Intercellular Junctions; Cell Tracking; [SDV.EE.SANT]Life Sciences [q-bio]/Ecology; [SDV.EE.ECO]Life Sciences [q-bio]/Ecology; [SDV.EE.IEO]Life Sciences [q-bio]/Ecology; environment/Ecosystems; [SDV.AEN]Life Sciences [q-bio]/Food and Nutrition; [SDV.EE.IEO]Life Sciences [q-bio]/Ecology, environment/Symbiosis; HeLa Cells

AbstractContact-dependent intercellular transfer (codeIT) of cellular constituents can have functional consequences for recipient cells, such as enhanced survival and drug resistance. Pathogenic viruses, prions and bacteria can also utilize this mechanism to spread to adjacent cells and potentially evade immune detection. However, little is known about the molecular mechanism underlying this intercellular transfer process. Here, we present a novel microscopy-based screening method to identify regulators and cargo of codeIT. Single donor cells, carrying fluorescently labelled endocytic organelles or proteins, are co-cultured with excess acceptor cells. CodeIT is quantified by confocal microscopy and image analysis in 3D, preserving spatial information. An siRNA-based screening using this method revealed the involvement of several myosins and small GTPases as codeIT regulators. Our data indicates that cellular protrusions and tubular recycling endosomes are important for codeIT. We automated image acquisition and analysis to facilitate large-scale chemical and genetic screening efforts to identify key regulators of codeIT.