Zelladhäsion; Crosstalk; Kraftspektroskopie; Zellbiologie; Zelladhäsionsrezeptoren
Kragl Martin, Schbert Rajib, Karsjens Haiko, Otter Silke, Bartosinska Barbara, Jeruschke Hay, Weiss Jürgen, Chen Chunguang, Alsteens David, Kuss Oliver, Speier Stephan, Eberhard Daniel, Müller Daniel J., Lammert Eckhard (2016), The biomechanical properties of an epithelial tissue determine the location of its vasculature, in Nature Communications
, 7, 13560.
Yu Miao, Wang Jinghe, Muller Daniel J., Helenius Jonne (2015), In PC3 prostate cancer cells ephrin receptors crosstalk to β1-integrins to strengthen adhesion to collagen type I, in Scientific Reports
, 5, 8206.
Miao Yu, Strohmeyer Nico, Wang Jinghe, Muller Daniel J., Helenius Jonne (2015), Increasing throughput of AFM-based single cell adhesion measurements through multisubstrate surfaces, in Beilstein J. Nanotechnol
, 6, 157-166.
Sorce Barbara, Escobedo Carlos, Toyoda Yusuke, Stewart Martin P., Cattin Cedric J., Newton Richard, Banerjee Indranil, Stettler Alexander, Roska Botond, Eaton Suzanne, Hyman Anthony A., Hierlemann Andreas, Müller Daniel J. (2015), Mitotic cells contract actomyosin cortex and generate pressure to round against or escape epithelial confinement, in Nature Communications
, 6, 8872.
Te Riet Joost, Helenius Jonne, Strohmeyer Nico, Cambi Alessandra, Figdor Carl G, Müller Daniel J (2014), Dynamic coupling of ALCAM to the actin cortex strengthens cell adhesion to CD6., in Journal of cell science
, 127(Pt 7), 1595-1606.
Friedrichs Jens, Legate Kyle R, Schubert Rajib, Bharadwaj Mitasha, Werner Carsten, Müller Daniel J, Benoit Martin (2013), A practical guide to quantify cell adhesion using single-cell force spectroscopy., in Methods (San Diego, Calif.)
, 60(2), 169-178.
Beckmann Jan, Schubert Rajib, Chiquet-Ehrismann Ruth, Müller Daniel J (2013), Deciphering teneurin domains that facilitate cellular recognition, cell-cell adhesion, and neurite outgrowth using atomic force microscopy-based single-cell force spectroscopy., in Nano letters
, 13(6), 2937-2946.
Friedrichs Jens, Werner Carsten, Müller Daniel J (2013), Quantifying cellular adhesion to covalently immobilized extracellular matrix proteins by single-cell force spectroscopy., in Methods in molecular biology (Clifton, N.J.)
, 1046, 19-37.
Teräväinen Terhi P, Myllymäki Satu M, Friedrichs Jens, Strohmeyer Nico, Moyano Jose V, Wu Chuanyue, Matlin Karl S, Muller Daniel J, Manninen Aki (2013), αV-integrins are required for mechanotransduction in MDCK epithelial cells., in PloS one
, 8(8), 71485-71485.
Schubert Rajib, Strohmeyer Nico, Bharadwaj Mitasha, Ramanathan Subramanian P, Krieg Michael, Friedrichs Jens, Franz Clemens M, Muller Daniel J, Assay for characterizing the recovery of vertebrate cells for adhesion measurements by single-cell force spectroscopy., in FEBS letters
Bharadwaj Mitasha, Strohmeyer Nico, Colo Georgina P., Helenius Jonne, Beerenwinkel Niko, Schiller Herbert B., Fässler Reinhard, Müller Daniel J., aV-class integrins exert dual roles on a5b1 integrins to strengthen adhesion to fibronectin, in Nature Communications
, 8, 14348.
Cells use specific and controllable adhesion mechanisms to interact with substrates, cells and tissue. Although most of the receptors involved in these adhesion mechanisms are known, the mechanisms by which they regulate adhesion remain largely unknown. The primary reason for this is that conventional cell adhesion assays are not suited to provide quantitative insights on mechanisms that regulate cell adhesion. Single-cell force spectroscopy (SCFS) allows the quantitative characterization of cellular adhesion mechanisms down to the contribution of single cell adhesion molecules (CAMs). We intend to expand and systematically implement SCFS to capture the broad mechanisms regulating the CAMs of living cells. Cells express and regulate CAMs to control whether or not, how strong and how long they adhere to surfaces. CAMs are receptors that form adhesive interactions, activate signal transduction pathways and regulate cellular processes. In regulatory processes termed ‘crosstalk’, CAMs regulate the activity of other CAMs. This allows the cell to actively adapt its adhesion properties to the environment. We have recently introduced SCFS to characterize such crosstalk in HeLa cells (Friedrichs et al., 2010b). For the first time, we observed that and quantified to which extend collagen type I-binding to a1ß1 integrin regulates cell adhesion mediated by a5ß1 integrin to fibronectin. Perturbation experiments showed that this crosstalk is unidirectional from a1ß1 to a5ß1 integrin and functioned by endocytosis of a5ß1 integrin. So far no systematic study or screen has been developed to identify which receptors crosstalk with CAMs and to quantify the extent this crosstalk modulates the adhesion of CAMs. Therefore, we will implement SCFS to systematically screen for crosstalk regulating adhesion of various CAMs in different human and vertebrate cells. In addition to crosstalk between CAMs, the implemented SCFS assay will allow us to determine how extracellular signals such as growth factors regulate adhesive properties of cells. The SCFS-based screening will focus on proteins known to induce cellular responses. Therefore, we will collaborate with several research groups that have a broad expertise on different extracellular matrix (ECM) proteins and CAMs, and in the molecular cell biology and genetics of the cellular systems. The collaborators will provide us with various substrates, cells and genetic tools (i.e. RNAi constructs, GFP-tagged receptors and CAMs, molecular compounds, specific blocking antibodies) to manipulate cellular system and CAMs. This unique combination of cellular systems, and of molecular cell biological and genetic tools will be essential to examine the mechanisms of the crosstalk networks discovered. Furthermore, the SCFS-based experiments will be performed in combination with advanced optical microscopy (i.e. fluorescence, confocal, TIRF, structured illumination, spinning disc) to provide structural insights into cellular mechanisms regulating crosstalk. This unique combination of nano- and microscopic tools will allow us to study to which extent the dynamic reassembly of the actomyosin cortex or of cell surface receptors modulates the adhesion of a given CAM. While our SCFS-based screens will focus on a wide array of cell surface receptors and cellular systems, it will provide a general unbiased array of ECM proteins that induce crosstalk with CAMs and of intercellular signaling proteins that are involved in this crosstalk. This project will not only be the first systematic quantitative characterization of the adhesive state of cells and its regulation by extracellular factors, but include follow up examination of specific mechanisms of the crosstalks discovered.