Centre staff
Dr Rowena Bunning
Principal lecturer in cell biology
Phone 0114 225 3012
Email r.a.bunning@shu.ac.uk
Research interests
Extracellular matrix breakdown and repair
Research is focussed on investigating the mechanisms of extracellular matrix (ECM) breakdown and repair with particular reference to the roles of proteinases and cytokines in this process. This is particularly relevant to cartilage breakdown in arthritis
and degenerative disc disease (DDD),
and also remodelling of central nervous system ECM, as occurs in multiple sclerosis (MS) and stroke. An understanding of mechanisms and control of ECM breakdown may lead to the development of new, more effective drugs for the treatment of arthritis, DDD and MS.
The effects of cannabinoids on chondrocyte metabolism
In addition to their well characterised psychotic effects, cannabinoids demonstrate analgesic, anti-inflammatory and immunosuppressive properties and have been observed to reduce joint damage in animal models of arthritis. We are interested in determining whether cannabinoids have any direct action on chondrocytes, which may be chondroprotective and account for some of their observed anti-arthritic effects. Our studies have shown inhibition of cytokine-stimulated nitric oxide production by cannabinoids R-(+)-win 55, 212-2 (win-55) and HU210 and inhibition of cytokine-stimulated cartilage proteoglycan and collagen breakdown. This work is in collaboration with Professor Kim Rainsford at Sheffield Hallam University
and is being continued in collaboration with Dr Christine Le Maitre, Sheffield Hallam University and Dr Aileen Crawford, University of Sheffield. It is also being extended to investigate the effects of cannabinoids on the chondrocyte-like cells from intervertebral discs. These studies will enable us to evaluate the potential of cannabinoids as therapeutic agents for arthritis and DDD. .
The role of ADAM 17 in the pathogenesis of multiple sclerosis (MS)
ADAM 17 (a disintegrin and metalloproteinase) is a sheddase for proteins involved in inflammation, including tumour necrosis factor (TNF) which is implicated in the pathogenesis of MS. We have studied the localisation of ADAM 17 in normal central nervous system (CNS) and MS CNS by immunohistochemistry. In normal CNS, ADAM 17 is expressed by astrocytes and endothelial cells, suggesting a role in normal CNS pathology. In inflammatory MS lesions, ADAM 17 appears increased in astrocytes and macrophage/microglial cells, suggesting a role in MS pathology. Studies of ADAM 17 in spinal cords in an animal model of MS in rats, experimental autoimmune encephalomyelitis (EAE), have also indicated increased expression of ADAM 17 in astrocytes and inflammatory cells in peak disease. This work is in collaboration with Professor Nicola Woodroofe.
Of particular interest is the functional significance of ADAM17 in MS. Studies using human brain endothelial cells as a model of the blood brain barrier (BBB) have shown that ADAM17 is not responsible for the shedding of fractalkine, a chemokine and adhesion molecule which may modulate the passage of inflammatory cells through the BBB and into the CNS. This work was in collaboration with Professor Nicola Woodroofe and Dr Basil Sharrack, University of Sheffield. We are also currently investigating the functional significance of ADAM17 using ADAM17 knockdown. Adeno-associated viral vectors encoding shRNA targeting ADAM17 are being used to knockdown ADAM17 in a Biozzi mouse model of EAE. This work is being funded by the Multiple sclerosis Society of Great Britain and Northern Ireland in collaboration with Professor Nicola Woodroofe, Prof Mimoun Azzouz, University of Sheffield and Dr Chris Bolton, Queen Mary's School of Medicine and Dentistry, University of London.
The role of ADAMTS-1, -4, -5 and -9 in the pathogenesis of multiple sclerosis and stroke
The ADAMTS (a disintegrin and metalloproteinase with thrombospondin type motif) group of enzymes are related to the ADAMs but are secreted enzymes, lacking transmembrane and cytoplasmic domains, instead having one or more thrombospondin motifs. ADAMTS -1, 4 and -5 are aggrecanases thought to be the main enzymes responsible for aggrecan breakdown in cartilage. The brain also has aggrecan-like proteoglycans in its ECM therefore these ADAMTSs may be involved in ECM breakdown in MS and stroke, allowing infiltration of damaging inflammatory cells. ADAMTS-1, -4 and -5 mRNA and protein have been demonstrated to be present in normal and MS white matter. A key finding has been an increase of ADAMTS-4 in MS lesions suggesting a role in MS pathogenesis. Conversely, studies of these enzymes in EAE demonstrated a decrease in ADAMTS-4 mRNA and protein.
Studies using the transient middle cerebral artery occlusion model of stroke in rats, in collaboration with Dr S Allen (Manchester) have shown an increase in mRNA expression of ADAMTS-1, -4, -5 and -9 in the occluded hemisphere, suggesting a role in stroke pathogenesis. Work on MS and stroke is in collaboration with Professor Nicola Woodroofe at Sheffield Hallam University and Dr David Buttle at Sheffield University and was funded by The Wellcome Trust.
Work on the functional significance of ADAMTSs in MS is continuing, in particular the presence of versican neoepitopes produced by ADAMTS cleavage in normal and MS tissue is being investigated as an indicator of in vivo ADAMTS activity. This work is in collaboration with Dr Alison Cross and Dr Gail Haddock, Sheffield Hallam University and Dr David Buttle, University of Sheffield.
Studies on the role of ADAM17 and ADAMTS-1, -4 and -5 in metastatic liver disease and liver degeneration.
Production of proteolytic enzymes is important in processes required for tumour growth and metastasis, in particular enzymes involved in ECM breakdown and in cytokine shedding. Production of ADAM17 and ADAMTS-1, -4 and -5 was studied in human hepatocellular carcinoma cell lines HepG2 and HuH7 and LX-2 a hepatic stellate cell line. All the cell lines expressed these enzymes and their endogenous inhibitor TIMP-3. Their expression could be differentially modulated by pro-inflammatory cytokines which are produced during tumour development (IL-1β, TNFα and IL-6).
Fractalkine may be involved in recruitment of immune cells to tumours where they may have anti-tumour activities. Conversely, fractalkine may promote angiogenesis and tumour growth. Fractalkine production and shedding was investigated in response to IL-1β and TNFα in HepG2 cells. These cytokines increased fractalkine shedding, however ADAM17 was not responsible for fractalkine cleavage as down-regulation with specific siRNA did not reduce shedding. This work was done in collaboration with Dr Maria Blair at Sheffield Hallam University and Drs Nigel Bird and David Mangnall, University of Sheffield
Collaborators
- Dr Stuart Allen, University of Manchester
- Professor Mimoun Azzouz, University of Sheffield
- Dr Nigel Bird, University of Sheffield
- Dr Chris Bolton, Queen Mary's School of Medicine and Dentistry, University of London.
- Dr David Buttle, University of Sheffield
- Dr Aileen Crawford, University of Sheffield
- Dr David Mangnall, University of Sheffield
- Dr Basil Sharrock, University of Sheffield
- Mark Wilkinson, University of Sheffield and Northern General Hospital
Recent publications
Turner SL, Mangnall D, Bird NC, Blair-Zadjel M, Bunning RAD (2010) Effects of pro-inflammatory cytokines on the production of soluble fractalkine and ADAM17 by hepG2 cells. Journal of Gastrointestinal and Liver Disease 19, 265-271.
Hurst LA, Bunning RAD, Couraud P-O, Romero IA, Weksler BB, Sharrack B, Woodroofe MN (2009) Expression of ADAM-17, TIMP3 and fractalkine in the human brain endothelial cell line, hCMEC/D3, following pro-inflammatory cytokine treatment. Journal of Neuroimmunology 210, 108-112.
SL Turner, ME Blair-Zajdel and RAD Bunning (2009) ADAMs and ADAMTSs in Cancer. The British Journal of Biomedical Science 66, 117-127.
Reid MJ, Cross AK, Haddock G, Allan SM, Stock CJ, Woodroofe MN, Buttle DJ, Bunning RAD (2009). ADAMTS-9 expression is up-regulated following transient middle cerebral artery occlusion (tMCAo) in the rat. Neuroscience Letter 452: 252-257.
Haddock G, Cross AK, Allan S, Sharrack B, Callaghan J, Bunning RAD, Buttle DJ, Woodroofe MN (2007) Brevican and phosphacan expression and localisation following transient middle cerebral artery occlusion in the rat. Biochemistry Society Transactions 35, 692-694.
Mbvundula EC, Bunning RAD, Rainsford KD (2006) Arthritis and cannabinoids: HU-210 and win-55,212-2 prevent IL-1α-induced matrix degradation in bovine articular chondrocytes in vitro. Journal of Pharmacy and Pharmacology 58, 351-358.
Cross A, Haddock G, Stock CJ, Allan S, Surr J, Bunning RAD, Buttle DJ, Woodroofe MN (2006) ADAMTS-1 and -4 are up-regulated following transient middle cerebral artery occlusion in the rat and their expression is modulated by TNF in cultured astrocytes. Brain Research 1088, 19-30.
Cross A, Haddock G, Stock CJ, Allan S, Surr J, Bunning RAD, Buttle DJ, Woodroofe MN (2006) ADAMTS-1 and -4 are up-regulated following transient middle cerebral artery occlusion in the rat and their expression is modulated by TNF in cultured astrocytes. Brain Research 1088, 19-30.
Haddock G, Cross AK, Surr J, Plumb J, Buttle DJ, Bunning RAD, Woodroofe MN (2006) Expression of ADAMTS-1, -4, -5 and TIMP-3 in normal and multiple sclerosis CNS white matter. Multiple Sclerosis 12, 386-389.
Cross AK, Haddock G, Surr J, Plumb J, Smith T, Bunning RAD, Buttle DJ, Woodroofe MN (2006) Differential expression of ADAMTS -1, -4, -5, TIMP-3 and chondroitin sulphate proteoglycans in rat spinal cord at different stages of acute experimental autoimmune encephalomyelitis. Journal of Autoimmunity 26, 16-23.
Plumb J, McQuaid S, Cross AK, Surr J, Haddock G, Bunning RAD, Woodroofe MN (2006) Upregulation of ADAM-17 expression in active lesions in multiple sclerosis. Multiple Sclerosis 12, 375-385.
Mbvundula EC, Bunning RAD, Rainsford KD (2005) Effects of cannabinoids on nitric oxide production by chondrocytes and proteoglycan degradation in cartilage. Biochemical Pharmacology 69, 635-640.
Plumb J, Cross AK, Surr J, Haddock G, Smith T, Bunning RAD, Woodroofe MN (2005). ADAM-17 and TIMP3 protein and mRNA expression in spinal cord white matter of rats with acute experimental autoimmune encephalomyelitis. J Neuroimmunology 164, 1-9.
Mbvundula EC, Rainsford KD, Bunning RAD (2004) Cannabinoids in pain and inflammation. Inflammopharmacology 12(2): 99-114.
Rainsford KD, Omar H, Ashraf A, Hewson AT, Bunning RAD, Rishiraj R, Shepherd P, Seabrook RW (2002) Recent pharmacodynamic and pharmacokinetic findings on oxaprozin. Inflammopharmacology 10 (3), 185-239.
Goddard DR, Bunning RAD, Woodroofe MN (2001) Astrocyte and endothelial cell expression of ADAM 17 (TACE) in adult human CNS. Glia 34, 267-271.
Book publications
Woodroofe NM, Bunning RAD (2005) Proteases and peptidases in EAE, Chapter A19, In:Experimental models of Multiple Sclerosis, Eds E Lavi, CS Constantinescu, Springer Science + Business media Inc, New York, pp 391 - 413.
PhD students
- Gehan Gibrel - the functional roles of ADAM-17 and ADAMTS-1, -4 and -5 in Multiple Sclerosis
- Sara Dunn - The effects of cannabinoids on chondrocyte metabolism
