Professor Steve Winder

Steve WinderProfessor of Molecular Cell Biology
Director of Postgraduate Teaching
Department of Biomedical Science
The University of Sheffield
Western Bank
Sheffield S10 2TN
United Kingdom

email: s.winder@sheffield.ac.uk
Tel: +44 (0) 114 222 2332
Room: B2 06 Florey building

General

Career history

  • Present: Professor of Molecular Cell Biology, Department of Biomedical Science, Department, University of Sheffield, Sheffield UK
  • 2003-2005: Reader, Department of Biomedical Science, University of Sheffield, Sheffield UK
  • 1999-2003 : Lecturer (99-01) Reader (02-03), University of Glasgow
  • 1995-1999 : Wellcome Trust Fellow, University of Edinburgh,
  • 1992-1995 : Staff Scientist, Laboratory of Molecular Biology, Cambridge - Advisor, Jake Kendrick-Jones
  • 1988-1992: Postdoctoral Fellow, Biochemistry, University of Calgary, Advisor - Mike Walsh.
  • 1994-1998 : PhD, University of Reading, Supervisor - Isabel Forsyth

Research interests

The main research focus of my group is centered around understanding the normal functions of the cell adhesion and signalling adaptor protein dystroglycan and in diseases such as muscular dystrophy and cancer. In particular we are investigating the regulation of the actin cytoskeleton, adhesion mediated signalling and cell motility and invasion using molecular cell biology and transgenic approaches.

My research group is part of the Centre for Membrane Interactions and Dynamics (CMIAD).

CMIAD

Activities and distinctions

  • Member of Editorial Boards of: International Journal of Cell Biology, Journal of Biomedicine and Biotechnology, Advanced Studies in Biology, Landes Bioscience, also member of Biochemical Journal Editorial Advisory Panel, and Board of Protein Modules Consortium.
  • Catalan Agency for Health Technology Assessment and Research grant review committee member. Regular grant reviewer for BBSRC, Welcome Trust, Telethon Italia and Asociation Française Contre les Myopathies.
  • Invited talks at FASEB Summer Research Conference Snowmass USA; EMBO Workshop on 'Coiled coils collagen and co-proteins' Alpbach, Austria; ESF Conference on ‘Functional Protein Modules’ Seefeld, Austria; British Society for Cell Biology Annual Meeting, York, England.
  • Invited peer reviewed reviews in Trends in Cell Bioloy, Current Opinion in Cell Biology and Trends in Biochemical Science. Editor and author of Molecular Mechanisms of Muscular Dystrophies, Landes Bioscience.

Funding

  • Duchenne Parent Project NL.
  • Action Duchenne. University of Sheffield

Selected publications since 2014

Full publications list

Research

The laminin binding protein dystroglycan plays multiple roles in cell adhesion, signalling and membrane cytoskeleton stability. Perturbation of dystroglycan function underlies several muscular dystrophies and is also a secondary consequence of adenocarcinoma progression. Changes to the post-translational modification of dystroglycan are crucial in directing the associations, cellular localisation and ultimately degradation of dystroglycan. Our aim is to elucidate the mechanisms and consequences of these post-translational modifications in order to better understand dystroglycan function and to identify potential therapeutic targets for the treatment of muscular dystrophy or cancer.

We employ in vitro, in/ex vivo fish and mouse genetic models with clinically relevant archival tissue samples or immortalised cell lines. Dystroglycan function is dissected through the use of molecular cell biology approaches, and potential therapeutic targets are assessed in vitro and in vivo. Recently we have developed a novel therapeutic approach for the treatment of Duchenne muscular dystrophy using inhibitors of tyrosine phosphorylation and proteasomal degradation. Through the use of zebrafish screening and phenotypic analysis in mdx mice and human DMD myoblasts we are in the process of validating the potential for repurposed drugs as a precursor to initiating clinical trials. Physiological analysis is carried out I collaboration with Nic Wells at the RVC London.

In vitro models of prostate cancer have revealed a role for the post-translational proteolytic processing and nuclear targeting of dystroglycan. Current efforts are centred around characterising a role as part of the LINC complex in the inner nuclear membrane. These studies form part of an ongoing collaboration with Bulmaro Cisneros, CINVESTAV Mexico City.

SJW_Research

Positions Available

Postgraduate PhD Opportunities

1. The Dystroglycan LINC: the functions of dystroglycan in the nuclear envelope in muscular dystrophy and cancer

Co-supervisor: Professor Jamie Hobbs (Physics and Astronomy)

Background

Biological membranes in animals have little intrinsic resistance to mechanical forces. Instead support is provided by membrane associated protein complexes. The plasma membrane, mitochondrial membrane and nuclear membrane all possess distinct integral membrane protein complexes that connect to the cytoskeleton and protect and position the membrane within the cell. We have found recently that dystroglycan, a plasma membrane adhesion receptor, characterised primarily for its role in membrane of striated muscle is also present in the in the nuclear envelope.  Moreover, we have found that dystroglycan depletion leads to disruption of the nuclear membrane.

Objectives:

  1. To determine how dystroglycan interacts with specific nuclear envelope protein complexes, such as the LINC complex and lamins.
  2. To assess the contribution of dystroglycan to the mechanical properties and physical integrity of the nuclear envelope during cell migration in two and three dimensions
  3. Evaluate the contribution of dystroglycan to spatial and mechanical responses to nuclear deformation.

Novelty and Timeliness

Dystroglycan is the only example of a transmembrane adhesion receptor that has structural and mechanical roles in plasma membrane and nuclear membrane. Through cytoskeletal connections it provides support to both membrane structures in tissues like muscle that are subjected to continued physical stress. The Winder lab and collaborators in Mexico are the only groups worldwide working on this novel aspect of dystroglycan function.

Experimental Approach

This is an integrated multidisciplinary project involving molecular cell biology, biophysics and biomechanics. The student will use these approaches in normal and dystroglycan knock-out myoblasts, including morphological analysis by immunofluorescence and analysis of interactions by immunoprecipitation. Migration of cells in 2D and 3D microfluidic chambers will assess the mechanical properties of the nucleus to deformation, whilst atomic force microscopy will assess the contribution of dystroglycan to the biophysical properties of isolated nuclei and nuclei in situ.

Dystroglycan is an essential protein that is a linker through biological membranes including the plasma membrane and the nuclear membrane. This project will address fundamental aspects of nuclear structure and dynamics employing a multidisciplinary approach comprising molecular cell biology and advanced microscopy. The project will be supervised by two world leading experts in their fields: Professor Steve Winder in the Department of Biomedical Science who works on the cell biology of dystroglycan, and Professor Jamie Hobbs who is at the forefront of research advances using atomic force microscopy (AFM) to probe biological systems.

The project will use genetic approaches in cells in tissue culture to manipulate the function of dystroglycan, a newly identified component of the nuclear membrane, and how it contributes to nuclear shape, nuclear resistance to stress and nuclear position in the cell. We will use biophysical approaches to measure nuclear deformation and stability and cell biological approaches to investigate the effects of altered nuclear deformation on cell migration in two and three dimensions, using microfluidic chambers. AFM and high resolution fluorescence microscopy, coupled with genetic manipulation will provide a powerful combination of approaches to understand the essential functions of dystroglycan in the nucleus.

References:

  • IA Martínez-Vieyra, A Vásquez-Limeta, R González-Ramírez, SL Morales-Lázaro, M Mondragón, R Mondragón, A Ortega, SJ Winder & B Cisneros. (2013)
    BBA Mol Cell Res. 1833, 698-711.
    A role for b-dystroglycan in the organisation and structure of the nucleus in myoblasts.
  • D Leocadio-Victoria, A Mitchell & SJ Winder (2016)
    J Cell. Biochem. 117, 2149-57
    γ-secretase dependent nuclear targeting of dystroglycan.
  • G. Mathew, A.Mitchell, J.M.Down, F.C.Hamdy, C.Eaton, D.J.Rosario, S.S.Cross & S.J.Winder (2013)
    Sci. Rep. 3, 2792.
    Nuclear targeting of dystroglycan promotes the expression of androgen regulated transcription factors in prostate cancer.

2. Targeting dystroglycan to the nucleus in muscular dystrophy and cancer

Background

Dystroglycan is an essential cell adhesion receptor required for early embryonic development. Genetic loss of function gives rise to severe muscular dystrophies with neuronal involvement. Post-translational loss of function also occurs in Duchenne muscular dystrophy and in some cancers. This includes phosphorylation, proteolysis and ubiquitination. Moreover some proteolytic fragments of dystroglycan are targeted to the nucleus where they have effects on transcription. As part of our analysis of dystroglycan post-translational modifications, we identified a lipid modification - palmitoylation of a conserved cysteine residue that could act to anchor bioactive dystroglycan fragments to the membrane, both at the cell surface and in the nucleus.

Aims:

To examine the function of dystroglycan palmitoylation in cellular targeting and in cellular phenotypes associated with muscular dystrophy and cancer.

Techniques:

PCR-based site-directed mutagenesis, and cloning of dystroglycan mutants. Overexpression of dystroglycan mutants in tissue culture cells. Analysis of subcellular distribution of dystroglycan by quantitative immunofluorescence microscopy and cell fractionation. In vitro assays of cell invasion and metastatic growth.

References:

  • Leocadio D, Mitchell A, Winder SJ.
    γ-Secretase Dependent Nuclear Targeting of Dystroglycan.
    J Cell Biochem. 2016 Mar 18. doi: 10.1002/jcb.25537
  • Martínez-Vieyra IA, Vásquez-Limeta A, González-Ramírez R, Morales-Lázaro SL, Mondragón M, Mondragón R, Ortega A, Winder SJ, Cisneros B.
    A role for β-dystroglycan in the organization and structure of the nucleus in myoblasts.
    Biochim Biophys Acta. 2013 1833:698-711
  • G. Mathew, A.Mitchell, J.M.Down, F.C.Hamdy, C.Eaton, D.J.Rosario, S.S.Cross & S.J.Winder (2013)
    Nuclear targeting of dystroglycan promotes the expression of androgen regulated transcription factors in prostate cancer.
    Sci. Rep. 3, 2792.

For further information about these projects and how to apply, please see our PhD Opportunities page.

PhD Opportunities

Members

Miss Katherine Collins-Taylor
Postgraduate Student
Email: KLCollins-Taylor1@sheffield.ac.uk
Extension: 22306

Ms Tracy Emmerson
Research Associate
Email: t.lariviere@sheffield.ac.uk
Extension: 22306/22328

Miss Laura Jacobs
Postgraduate Student
Email: mdp12laj@sheffield.ac.uk
Extension: 22306

Dr Gemma Woodward
Postdoctoral Researcher
Email: g.woodward@sheffield.ac.uk
Extension: 22306