Metastasising Fibroblasts Show an HDAC6-Dependent Increase in Migration Speed and Loss of Directionality Linked to Major Changes in the Vimentin Interactome

• Published in: International journal of molecular sciences Vol. 23; no. 4; p. 1961
• Main Authors: Evans, Caroline A, Kim, Hyejeong Rosemary, Macfarlane, Sarah C, Nowicki, Poppy I A, Baltes, Carsten, Xu, Lei, Widengren, Jerker, Lautenschläger, Franziska, Corfe, Bernard M, Gad, Annica K B
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 10.02.2022; MDPI
• Subjects: Actin; Actins - metabolism; Animals; Cell adhesion & migration; Cell Adhesion - physiology; Cell cycle; cell migration; Cell Movement - physiology; Cell Shape - physiology; Cell size; Cell spreading; Cell-Matrix Junctions - metabolism; Cells, Cultured; Collagen; Collagen - metabolism; Cytoskeleton; Cytoskeleton - metabolism; Extracellular matrix; Fibroblasts; Fibroblasts - metabolism; Fibroblasts - pathology; genome-wide mass spectrometry; histone deacetylase 6; Histone Deacetylase 6 - metabolism; Humans; Keratin; Metastasis; Mice; Motility; oncogenes; Oncogenes - physiology; Parameter identification; Phenotypes; Proteins; Proteomics; stimulated emission depletion microscopy; Talin; Tropomyosin; Tumors; Vimentin; Vimentin - metabolism; vimentin interactome; fibroblasts; genome-wide mass spectrometry; histone deacetylase 6; metastasis; cell migration; oncogenes; vimentin; vimentin interactome; stimulated emission depletion microscopy
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms23041961

Metastasising cells express the intermediate filament protein vimentin, which is used to diagnose invasive tumours in the clinic. We aimed to clarify how vimentin regulates the motility of metastasising fibroblasts. STED super-resolution microscopy, live-cell imaging and quantitative proteomics revealed that oncogene-expressing and metastasising fibroblasts show a less-elongated cell shape, reduced cell spreading, increased cell migration speed, reduced directionality, and stronger coupling between these migration parameters compared to normal control cells. In total, we identified and compared 555 proteins in the vimentin interactome. In metastasising cells, the levels of keratin 18 and Rab5C were increased, while those of actin and collagen were decreased. Inhibition of HDAC6 reversed the shape, spreading and migration phenotypes of metastasising cells back to normal. Inhibition of HDAC6 also decreased the levels of talin 1, tropomyosin, Rab GDI β, collagen and emilin 1 in the vimentin interactome, and partially reversed the nanoscale vimentin organisation in oncogene-expressing cells. These findings describe the changes in the vimentin interactome and nanoscale distribution that accompany the defective cell shape, spreading and migration of metastasising cells. These results support the hypothesis that oncogenes can act through HDAC6 to regulate the vimentin binding of the cytoskeletal and cell-extracellular matrix adhesion components that contribute to the defective motility of metastasising cells.