Polarised differentiation of epithelial cells is critical for their normal function and loss of polarity is a major factor in epithelial-to-mesenchymal transition (EMT) and conversion to an invasive cancer state. It is well known that epithelial cell polarisation is dependent on relatively stable apico-basal microtubule arrays whereas loss of these arrays and dynamic microtubules are required for EMT and cell migration. Epithelial remodelling occurs during early stages of EMT when a cell switches from an apico-basal polarised to a migratory phenotype and this is associated with dramatic changes in microtubule dynamics and organisation. However, relatively little is known about the molecular mechanisms regulating microtubule dynamics and reorganisation during EMT. We have recently identified a microtubule associated protein that when overexpressed stimulates epithelial remodelling and migration. Our preliminary data show that overexpression of this protein in 3D MDCKII matrigel cultures leads to basal cell protrusions and formation of cell aggregates rather than cyst. The aims of this project are to use 3D in vitro cultures and invasion assays that mimic the in vivo epithelial architecture and migration conditions to analyse the role of this and related microtubule associated proteins in microtubule dynamics and reorganisation during epithelial remodelling in order to better understand the process of EMT. We will use a combination of 3D in vitro epithelial cyst and invasion models together with GFP- and RNAi-technology and immuno-labelling in conjunction with high-resolution widefield fluorescence, confocal, multi-photon and live time-lapse microscopy to pursue these aims. The findings from this proposal will advance our understanding of epithelial remodelling and potentially identify novel drug targets for treatment of highly invasive cancers.

Bellett, G., Carter, J. M., Keynton, J., Goldspink, D., James, C., Moss, D. K. and Mogensen, M.M.2009. Microtubule plus-end and minus-end capture at adherens junctions is involved in the assembly of apico-basal arrays in polarised epithelial cells. Cell Motil Cytoskeleton 66: 893–908.

Moss, D. K., Bellett, G., Carter, J. M., Liovic, M., Keynton, J., Prescott, A. R., Lane, E. B. and Mogensen, M. M. (2007). Ninein is released from the centrosome and moves bi-directionally along microtubules. J Cell Sci 120, 3064-74.

Kohlmaier, G, J Loncarek, X Meng, BF McEwen, M M Mogensen, A Spektor, B D Dynlacht, A Khodjakov and P Gonczy. 2009. Overly Long Centrioles and Defective Cell Division upon Excess of the SAS-4-Related Protein CPAP. Curr Biol. 19:1012-1018

Baker N, Sharpe P, Culley K, Otero M, Bevan D, Newham P, Barker W, Clements KM,  Langham CJ, Goldring MB, Gavrilović J. Dual regulation of metalloproteinase expression in chondrocytes by Wnt-1-inducible signaling pathway protein 3/CCN6. Arthritis Rheum. 2012 64:2289-99.

Singh RJ, Mason JC, Lidington EA, Edwards DR, Nuttall RK, Khokha R, Knauper V, Murphy G,Gavrilovic J. Cytokine stimulated vascular cell adhesion molecule-1 (VCAM-1) ectodomain release is regulated by TIMP-3.Cardiovasc Res. 2005 Jul 1;67(1):39-49. Epub 2005 Mar 29.