Curved adhesions mediate cell attachment to soft matrix fibres in 3D

Mammalian cells adhere to the extracellular matrix (ECM) and sense mechanical cues through integrin-mediated adhesions1, 2. Focal adhesions and related structures are the primary architectures that transmit forces between the ECM and the actin cytoskeleton. Although focal adhesions are abundant when cells are cultured on rigid substrates, they are sparse in soft environments that cannot support high mechanical tensions3. Here, we report a new class of integrin-mediated adhesions, curved adhesions, whose formation is regulated by membrane curvature instead of mechanical tension. In soft matrices made of protein fibres, curved adhesions are induced by membrane curvatures imposed by the fibre geometry. Curved adhesions are mediated by integrin ɑVβ5 and are molecularly distinct from focal adhesions and clathrin lattices. The molecular mechanism involves a previously unknown interaction between integrin β5 and a curvature-sensing protein FCHo2. We find that curved adhesions are prevalent in physiologically relevant environments. Disruption of curved adhesions by knocking down integrin β5 or FCHo2 abolishes the migration of multiple cancer cell lines in 3D matrices. These findings provide a mechanism of cell anchorage to natural protein fibres that are too soft to support the formation of focal adhesions. Given their functional importance for 3D cell migration, curved adhesions may serve as a therapeutic target for future development.

[1]  C. Bertozzi,et al.  Membrane curvature regulates the spatial distribution of bulky glycoproteins , 2022, Nature Communications.

[2]  B. Cui,et al.  Expansion Microscopy for Imaging the Cell-Material Interface. , 2022, ACS nano.

[3]  B. Cui,et al.  Exploring cell surface-nanopillar interactions with 3D super-resolution microscopy , 2021, bioRxiv.

[4]  E. Ruoslahti,et al.  Tumor-penetrating therapy for β5 integrin-rich pancreas cancer , 2021, Nature Communications.

[5]  Allister F. McGuire,et al.  Membrane curvature underlies actin reorganization in response to nanoscale surface topography , 2019, Proceedings of the National Academy of Sciences.

[6]  Ge-fei Guan,et al.  Integrin Beta 5 Is a Prognostic Biomarker and Potential Therapeutic Target in Glioblastoma , 2019, Front. Oncol..

[7]  M. Radmacher,et al.  Nano-mechanical mapping of interdependent cell and ECM mechanics by AFM force spectroscopy , 2019, Scientific Reports.

[8]  M. Humphries,et al.  Clathrin-containing adhesion complexes , 2019, The Journal of cell biology.

[9]  Pere Roca-Cusachs,et al.  Integrins as biomechanical sensors of the microenvironment , 2019, Nature Reviews Molecular Cell Biology.

[10]  Allister F. McGuire,et al.  A nanostructure platform for live-cell manipulation of membrane curvature , 2019, Nature Protocols.

[11]  R. Fässler,et al.  Integrin activation by talin, kindlin and mechanical forces , 2019, Nature Cell Biology.

[12]  M. Lakadamyali,et al.  Reticular adhesions are a distinct class of cell-matrix adhesions that mediate attachment during mitosis , 2018, Nature Cell Biology.

[13]  A. Sonnenberg,et al.  Mechanisms of integrin αVβ5 clustering in flat clathrin lattices , 2018, Journal of Cell Science.

[14]  T. Betz,et al.  Frustrated endocytosis controls contractility-independent mechanotransduction at clathrin-coated structures , 2018, Nature Communications.

[15]  Andrew D. Doyle.  Fluorescent Labeling of Rat-tail Collagen for 3D Fluorescence Imaging. , 2018, Bio-protocol.

[16]  M. Rief,et al.  Multiplexing molecular tension sensors reveals piconewton force gradient across talin-1 , 2017, Nature Methods.

[17]  Benjamin Klapholz,et al.  Talin – the master of integrin adhesions , 2017, Journal of Cell Science.

[18]  S. Vassilopoulos,et al.  Tubular clathrin/AP-2 lattices pinch collagen fibers to support 3D cell migration , 2017, Science.

[19]  Francesca Santoro,et al.  Nanoscale manipulation of membrane curvature for probing endocytosis in live cells. , 2017, Nature nanotechnology.

[20]  Devin P. Sullivan,et al.  A subcellular map of the human proteome , 2017, Science.

[21]  Cheng Zhu,et al.  Mechanical regulation of a molecular clutch defines force transmission and transduction in response to matrix rigidity , 2016, Nature Cell Biology.

[22]  Kenneth M. Yamada,et al.  Mechanosensing via cell-matrix adhesions in 3D microenvironments. , 2016, Experimental cell research.

[23]  M. Sheetz,et al.  Integrin-beta3 clusters recruit clathrin-mediated endocytic machinery in the absence of traction force , 2015, Nature Communications.

[24]  M. Rief,et al.  Extracellular rigidity sensing by talin isoform–specific mechanical linkages , 2015, Nature Cell Biology.

[25]  Adam Byron,et al.  Definition of a consensus integrin adhesome and its dynamics during adhesion complex assembly and disassembly , 2015, Nature Cell Biology.

[26]  Clare M. Waterman,et al.  Integration of actin dynamics and cell adhesion by a three-dimensional, mechanosensitive molecular clutch , 2015, Nature Cell Biology.

[27]  Brian D. Slaughter,et al.  The membrane-associated proteins FCHo and SGIP are allosteric activators of the AP2 clathrin adaptor complex , 2014, eLife.

[28]  Andrei V Bakin,et al.  Integrin β5 contributes to the tumorigenic potential of breast cancer cells through Src-FAK and MEK-ERK signaling pathways , 2012, Oncogene.

[29]  Z. Kam,et al.  Fibroblast polarization is a matrix-rigidity-dependent process controlled by focal adhesion mechanosensing , 2011, Nature Cell Biology.

[30]  Kenneth M. Yamada,et al.  Cell-matrix adhesions in 3D. , 2011, Matrix biology : journal of the International Society for Matrix Biology.

[31]  Brenton D. Hoffman,et al.  Dynamic molecular processes mediate cellular mechanotransduction , 2011, Nature.

[32]  Kenneth M. Yamada,et al.  Direct comparisons of the morphology, migration, cell adhesions, and actin cytoskeleton of fibroblasts in four different three-dimensional extracellular matrices. , 2011, Tissue engineering. Part A.

[33]  S. Keeney,et al.  References and Notes Supporting Online Material Materials and Methods Figs. S1 to S5 Tables S1 and S2 References Movie S1 Fcho Proteins Are Nucleators of Clathrin-mediated Endocytosis , 2022 .

[34]  Stephanie I. Fraley,et al.  A distinctive role for focal adhesion proteins in three-dimensional cell motility , 2010, Nature Cell Biology.

[35]  R. Fässler,et al.  The Tail of Integrins, Talin, and Kindlins , 2009, Science.

[36]  Samuel J. Lord,et al.  Three-dimensional, single-molecule fluorescence imaging beyond the diffraction limit by using a double-helix point spread function , 2009, Proceedings of the National Academy of Sciences.

[37]  Adam Byron,et al.  Integrin ligands at a glance , 2006, Journal of Cell Science.

[38]  M. Sheetz,et al.  Local force and geometry sensing regulate cell functions , 2006, Nature Reviews Molecular Cell Biology.

[39]  R. Thompson,et al.  Quantitation of vitronectin in serum: evaluation of its usefulness in routine clinical practice. , 1993, Journal of Clinical Pathology.

[40]  Y. Takada,et al.  Distinct functions of integrin alpha and beta subunit cytoplasmic domains in cell spreading and formation of focal adhesions , 1993, The Journal of cell biology.

[41]  M. Hayashi,et al.  Activation of the collagen-binding of endogenous serum vitronectin by heating, urea and glycosaminoglycans. , 1992, Biochimica et biophysica acta.

[42]  D. Kirchhofer,et al.  Calcium as a potential physiological regulator of integrin-mediated cell adhesion. , 1991, The Journal of biological chemistry.

[43]  E. Engvall,et al.  Interaction of vitronectin with collagen. , 1986, The Journal of biological chemistry.

[44]  E. Ruoslahti,et al.  Serum spreading factor (vitronectin) is present at the cell surface and in tissues. , 1983, Proceedings of the National Academy of Sciences of the United States of America.

[45]  D. Gospodarowicz,et al.  Are factors originating from serum, plasma, or cultured cells involved in the growth‐promoting effect of the extracellular matrix produced by cultured bovine corneal endothelial cells? , 1983, Journal of cellular physiology.

[46]  A. Fountas,et al.  Plasma fibronectin in normal subjects and in various disease states. , 1981, Journal of clinical pathology.

[47]  G. Stoner,et al.  Histochemical demonstration of collagen fibers in ascorbic-acid-fed cell cultures , 1978, In Vitro.

[48]  Kristopher E Kubow,et al.  Reducing background fluorescence reveals adhesions in 3D matrices , 2012, Nature Cell Biology.

[49]  Stephanie I. Fraley,et al.  Reply: reducing background fluorescence reveals adhesions in 3D matrices , 2010, Nature Cell Biology.

[50]  B. Geiger,et al.  Environmental sensing through focal adhesions , 2009, Nature Reviews Molecular Cell Biology.