Professor of Molecular Carcinogenesis
Molecular regulation of transcription factor Nrf2 and its therapeutic manipulation to treat lung cancer and liver disease
Jacqui Wood Cancer Centre
University of Dundee
James Arrott Drive
Ninewells Hospital And Medical School
+(44) 01382 383182
Professor Hayes initially trained between 1972 and 1976 in the School of Biological Sciences at the University of Edinburgh, with final year of studies in Molecular Biology. He gained a PhD from the Medical School of the same university in 1980, awarded for investigation into bile acid-binding properties of hepatic glutathione S-transferases. In 1981, he was appointed a Lecturer within the University Department of Clinical Chemistry at Edinburgh. Here, he focused attention on the enzymology and protein chemistry of the glutathione S-transferase (GST) superfamily in rodents and human, and became particularly interested in the contribution of inducible class Alpha GST to chemoprevention against the liver carcinogen aflatoxin B1 and the contribution made by overexpressed GST in tumours to acquired resistance to anticancer drugs. In 1991, he took sabbatical leave from the University of Edinburgh to work in the laboratory of Dr Cecil B. Pickett, in the Department of Molecular Biology, Merck Frosst, Montreal, Canada, which was responsible for discovering the antioxidant response element (ARE) in the promoters of genes for inducible GST and for NAD(P)H:quinone oxidoreductase 1 (NQO1). Upon return from sabbatical, the University of Edinburgh promoted Dr Hayes to a Readership in Clinical Biochemistry.
In October 1992, Dr Hayes moved as a Reader to the University of Dundee to help Prof C. Roland Wolf set up the Biomedical Research Centre in the Medical School. In this environment, he focused more on the molecular biology and regulation of detoxication enzymes and identified a new family of inducible aldo-keto reductases (AKR) that metabolises a dialdehydic form of aflatoxin B1 and quinones. In January 1997, the University of Dundee promoted him to a personal chair. Since then, his interest in the role of the ARE in directing induction of detoxication genes has continued, and he has collaborated with Prof Masayuki Yamamoto (Tohoku, Japan) to demonstrate that the Nrf2 transcription factor regulates both basal and inducible expression of GST, AKR, NQO1 and glutathione biosynthetic enzymes. More recently, Prof Hayes’ lab has shown that control of the ARE-gene battery critically depends on the stability of Nrf2 protein, and inhibition of the ubiquitin ligase adaptor Keap1 blocks turn-over of the transcription factor. Workers in the Hayes laboratory were the first to show that Nrf2 stability is controlled by Keap1, and have more recently demonstrated the existence of three independent stress sensors in Keap1 that evolved separately.
Mechanisms of regulation of transcription factor Nrf2 and their relevance to chemoprotection and novel cancer treatment therapies.
The Nrf2 transcription factor is a master regulator of redox homeostasis. It is activated by diverse environmental agents and plays a fundamental role in cellular adaptation to oxidative stress and electrophilic chemicals. Specifically, Nrf2 controls the stress-inducible expression of a battery of genes for antioxidant proteins, NADPH regeneration enzymes, drug-metabolising enzymes, drug efflux pumps, and proteasome subunits. In normal cells, activation of Nrf2 has strong anti-inflammatory effects and limits damage to macromolecules caused by reactive oxygen species and electrophiles. Indeed, many cancer chemopreventive agents that protect against DNA damage by carcinogens do so by activating Nrf2 and inducing its target genes. On the other hand, Nrf2 is frequently constitutively activated in cancer cells, and this is associated with increased drug resistance and a higher rate of proliferation of such cells.
Given the involvement of Nrf2 in both the prevention of carcinogenesis in normal cells and in the promotion of tumourigenesis in cells in which cancer has been initiated, there is an obvious need to understand the mechanisms by which it is regulated. Nrf2 is principally controlled at the protein level through ubiquitylation, which targets the transcription factor for proteasomal degradation. To date, the ubiquitin ligase substrate adaptors Keap1 and beta-TrCP have been found to play central roles in repressing Nrf2 through their involvement in CRL(Keap1) and SCF(beta-TrCP) ubiquitin ligases: Keap1 regulates Nrf2 in an oxidative stress-sensitive fashion; beta-TrCP regulates Nrf2 in a glycogen synthase kinase-3 (GSK3)-dependent manner that also integrates several signal transduction pathways. Other mechanisms by which Nrf2 is regulated entail control of its subcellular localization.
The Hayes laboratory is particularly interested in how Keap1 and the beta-TrCP-GSK3 pathway sense stressors and whether such knowledge can be used to identify more potent chemopreventive agents that activate Nrf2 efficiently in normal cells, or to either discover small molecule inhibitors that repress Nrf2 in cancer cells. We have discovered that Keap1 contains three independent stress sensors and are characterizing them to determine how they are triggered, and how they might recover from modification by various stressors. Also, we are currently investigating the mechanisms by which GSK3 allows ubiquitylation of Nrf2 by SCF(beta-TrCP) and identify the signal transduction pathways that regulate phosphorylation of Nrf2 by GSK3. Biochemical, molecular biology and cell biology methods, along with transgenic mouse models, will be employed to address these questions.
- Hayes JD, Dinkova-Kostova AT. The Nrf2 regulatory network provides an interface between redox and intermediary metabolism. Trends Biochem Sci. 2014 Apr;39(4):199-218. Epub 2014 Mar 16.
- Lewerenz J, Baxter P, Kassubek R, Albrecht P, Liefferinge JV, Westhoff MA, Halatsch ME, Karpel-Massler G, Meakin PJ, Hayes JD, Aronica E, Smolders I, Ludolph AC, Methner A, Conrad M, Massie A, Hardingham GE, Maher P. Phosphoinositide 3-Kinases Upregulate System xc- via Eukaryotic Initiation Factor 2α and Activating Transcription Factor 4 - A Pathway Active in Glioblastomas and Epilepsy. Antioxid Redox Signal. 2014 Feb 6. [Epub ahead of print].
- Gan FF, Ling H, Ang X, Reddy SA, Lee SS, Yang H, Tan SH, Hayes JD, Chui WK, Chew EH (2013). A novel shogaol analog suppresses cancer cell invasion and inflammation, and displays cytoprotective effects through modulation of NF-κB and Nrf2-Keap1 signalling pathways. Toxicol Appl Pharmacol. 2013 Jul 27. doi:pii:S0041-008X(13)00314-1. 10.1016/j.taap.2013.07.011. [Epub ahead of print].
- Zhang Y, Ren Y, Li S, Hayes JD. Transcription Factor Nrf1 Is Topologically Repartitioned across Membranes to Enable Target Gene Transactivation through Its Acidic Glucose-Responsive Domains. PLoS One. 2014 Apr 2;9(4):e93458.
- Zhang Y & Hayes JD. (2013) The membrane-topogenic vectorial behaviour of Nrf1 controls its post-translational modification and transactivation activity. Sci Rep. 2013 Jun 18;3:2006. doi: 10.1038/srep02006.
- Wang H, Liu K, Geng M, Gao P, Wu X, Hai Y, Li Y, Li Y, Luo L, Hayes JD, Wang XJ, Tang X. (2013) RXRα Inhibits the NRF2-ARE Signaling Pathway through a Direct Interaction with the Neh7 Domain of NRF2. Cancer Res. 73: 3097-3108.
- Hourihan JM, Kenna JG, Hayes JD. (2013) The Gasotransmitter Hydrogen Sulfide Induces Nrf2-Target Genes by Inactivating the Keap1 Ubiquitin Ligase Substrate Adaptor Through Formation of a Disulfide Bond Between Cys-226 and Cys-613. Antioxid Redox Signal. 19: 465-481
- Chowdhry S, Zhang Y, McMahon M, Sutherland C, Cuadrado A & Hayes JD. (2013) Nrf2 is controlled by two distinct b-TrCP recognition motifs in its Neh6 domain, one of which can be modulated by GSK-3 activity. Oncogene 32: 3765-3781
- Hayes JD, Ashford ML (2012) Nrf2 orchestrates fuel partitioning for cell proliferation. Cell Metabolism 16: 139-141
- Bell KF, Al-Mubarak B, Fowler JH, Baxter PS, Gupta K, Tsujita T, Chowdhry S, Patani R, Chandran S, Horsburgh K, Hayes JD & Hardingham GE. (2011) Mild oxidative stress activates Nrf2 in astrocytes, which contributes to neuroprotective ischemic preconditioning. Proc. Natl. Acad. Sci. USA, 108(1):E1-2
- Rada P, Rojo AI, Chowdhry S, McMahon M, Hayes JD, Cuadrado A. (2011) SCF/b-TrCP promotes Glycogen synthase kinase-3-dependent degradation of the Nrf2 transcription factor in a Keap1-independent manner. Mol. Cell. Biol. 31: 1121-1133
- McMahon M, Lamont DJ, Beattie KA, Hayes JD (2010) Keap1 perceives stress via three sensors for the endogenous signaling molecules, nitric oxide, zinc and alkenals. Proc. Natl. Acad. Sci. USA, 107: 18838-18843
- Hayes JD, McMahon M, Chowdhry S, Dinkova-Kostova AT (2010) Cancer chemoprevention mechanisms mediated through the Keap1-Nrf2 pathway. Antiox. Redox Signal. 13: 1713-1748
- Chowdhry S, Nazmy MH, Meakin PJ, Dinkova-Kostova AT, Walsh SV, Tsujita T, Dillon JF, Ashford ML, Hayes JD (2010) Loss of Nrf2 markedly exacerbates non-alcoholic steatohepatitis. Free Rad. Biol. Med. 48: 357-371
- Kelleher MO, McMahon M, Eggleston IM, Dixon MJ, Taguchi K, Yamamoto M, Hayes JD (2009) 1-Cyano-2,3-epithiopropane is a novel plant-derived chemopreventive agent which induces cytoprotective genes that afford resistance against the genotoxic a,b-unsaturated aldehyde acrolein. Carcinogenesis 30: 1754-1762
- MacLeod AK, McMahon M, Plummer SM, Higgins LG, Penning TM, Igarashi K, Hayes JD (2009) Characterization of the cancer chemopreventive NRF2-dependent gene battery in human keratinocytes: demonstration that the KEAP1-NRF2 pathway, and not the BACH1-NRF2 pathway, controls cytoprotection against electrophiles as well as redox-cycling compounds. Carcinogenesis 30: 1571-1580
- Higgins LG, Kelleher MO, Eggleston IM, Itoh K, Yamamoto M, Hayes JD (2009) Transcription factor Nrf2 mediates an adaptive response to sulforaphane that protects fibroblasts in vitro against the cytotoxic effects of electrophiles, peroxides and redox-cycling agents. Toxicol. Appl. Pharmacol. 237: 267-280
- Hayes JD & McMahon M (2009) NRF2 and KEAP1 mutations: permanent activation of an adaptive response in cancer. Trends Biochem. Sci. 34: 176-188
- Wang XJ, Hayes JD, Henderson CJ, Wolf CR (2007) Identification of retinoic acid as an inhibitor of transcription factor Nrf2 through activation of retinoic acid receptor alpha. Proc. Natl. Acad. Sci. USA.104: 19589-19594
- McMahon M, Thomas N, Itoh K, Yamamoto M, Hayes JD (2006) Dimerisation of substrate adaptors can facilitate cullin-mediated ubiquitylation of proteins by a ‘tethering’ mechanism: A two-site interaction model for the Nrf2-Keap1 complex. J. Biol. Chem. 281: 24756-24768
Prof Hayes contributes to the MRes Cancer Biology course, the BMSc Pharmacology BSc Hons course and the SuperSeminar series that are run in the University of Dundee.
Kimimuepigha Ebesine (from Oct 2013): Project entitled ‘Inhibition of transcription factor Nrf2 by quassinoids and other natural products’.
Lauren Tebay (from Oct 2011): Project entitled ‘Mechanism of inhibition of inflammation by the antioxidant transcription factor Nrf2’.
Laura J. Brown (from Oct 2009): Thesis entitled ‘Regulation of Nrf2 through its Neh2 and Neh6 degrons’ [thesis submitted]
John M. Hourihan (2008-2012): Thesis entitled ‘Covalent modification of Keap1 by gasotransmitters as a mechanism to activate the transcription factor Nrf2’.
Sudhir Chowdhry (2008-2012): Thesis entitled ‘Regulation of Nrf2 and its contribution to prevention of inflammatory liver disease’.
Han Xiao (2006-2010): Thesis entitled ‘Mechanisms by which natural polyphenols regulate the expression of cytoprotective genes’.
A. Kenneth MacLeod (2004-2008): Thesis entitled ‘The antioxidant response of human keratinocytes’.
Yiguo Zhang (2003-2007): Thesis entitled ‘Molecular and cellular control of the Nrf1 transcription factor’.
Larry G. Higgins (2001-2006): Thesis entitled ‘Role of Nrf2 in adaptation to chemical stress’.
Timothy W.P. Devling (2001-2005): Thesis entitled ‘Cancer chemoprevention mechanisms in the human skin’.
Rachael E. Thomson (2001-2004): Thesis entitled ‘Mitochondrial glutathione S-transferase class Kappa: biochemical characterisation, localisation and response to toxic insult’.
Paul Nioi (2000-2003): Thesis entitled ‘Role of Nrf2 in regulation of induction of NAD(P)H:quinone oxidoreductase 1’
Ian R. Jowsey (1998-2001): Thesis entitled ‘Mammalian class Sigma glutathione S-transferases: catalytic properties and physiological functions’.
Christine Bonnesen (1998-2001): Thesis entitled ‘The antitumourigenic activity of the naturally occurring glucosinolates, glucobrassicin and neoglucobrassicin’.
Simon A. Chanas (1997-2001): Thesis entitled ‘The induction of drug detoxification and antioxidant enzymes in response to xenobiotics by transcription factor Nrf2’.
Cara Slattery (1997-2001): Thesis entitled ‘Regulation of the aflatoxin B aldehyde reductase gene by cancer chemopreventive agents’.
Anne M. Thomson (1996-1999): Thesis entitled ‘Role of prostaglandin-metabolising class Sigma glutathione S-transferase in both adaptive response to chemical stress and immune response’.
Vincent P. Kelly (1995-1999): Thesis entitled ‘Molecular characterization of multiple AFAR proteins which comprise a unique family of aldo-keto reductases’.
Tania O'Connor (1995-1999): Thesis entitled ‘Catalytic properties and regulation of human members of the aldo-keto reductase superfamily’.
Philip J. Sherratt* (1995-1998): Thesis entitled ‘Class Theta glutathione S-transferase and risk assessment for methylene chloride’.
Shan Zhong (1989-1992): Thesis entitled ‘Molecular genetic analysis of human mu class GST’, University College London.
Peter C. Hayes (1988-1993): Thesis entitled ‘Glutathione S-transferases in the pancreas’, University of Edinburgh.
Ian Meikle (1988-1991): Thesis entitled ‘Glutathione S-transferases in the adrenal cortex’, University of Edinburgh.
Amanda J. Hussey (1988-1991): Thesis entitled ‘Glutathione S-transferases: properties of mu-class and theta-class isoenzymes’, University of Edinburgh.
A. Forbes Howie (1986-1990): Thesis entitled ‘Measurement of human glutathione S-transferases by RIA’, University of Edinburgh.
Claire J. Wareing (1986-1990): Thesis entitled ‘The role of glutathione S-transferases in resistance to cytotoxic compounds’, University of Edinburgh.
Catherine Dolan (1985-1990): Thesis entitled ‘Regulation of mouse hepatic glutathione S-transferase’, University of Edinburgh.
Lesley I. McLellan (1984-1987): Thesis entitled ‘Cytosolic and microsomal isoenzymes of glutathione S-transferase’, University of Edinburgh.
Paul K. Stockman (1982-1985): Thesis entitled ‘The glutathione S-transferases in human liver cytosol’, University of Edinburgh.
Dr Colin J. Henderson (1981-1984): Thesis entitled ‘Studies on rat hepatic bile acid-binding proteins using photoaffinity techniques’, University of Edinburgh
Over the past 30 years, Prof Hayes has delivered at least 60 lectures at international meetings on various aspects of cancer chemoprevention, cancer drug resistance, the biochemistry of glutathione S-transferases, regulation of novel aldo-keto reductases and the role of transcription factor Nrf2 in pharmacology, which were held under the auspices of various scientific societies. The organizations that helped organise these conferences included the American Association for Cancer Research, the American Chemical Society, the American Institute for Cancer Research, the Biochemical Society, the British Toxicology Society, the European Society of Toxicology, the International Agency for Research on Cancer, the International Society for the Study of Xenobiotics, Japan Science and Technology Agency, Microsomes and Drug Oxidations, the National Institute of Environmental Health Sciences, the Physiological Society, the Royal Society of Edinburgh and the Swedish Chemical Society.
At present, Prof Hayes is helping to organise the ‘Environmental Response Symposium’ with Prof Masayuki Yamamoto (Tohoku, Japan), Prof Thomas W. Kensler (Pittsburgh, USA), Prof Donna D. Zhang (Arizona, USA) and Prof Steven R. Kleeberger (North Carolina, USA) to be held in Sendai, Japan on 28th February to 2nd March 2013.
Prof Hayes is also helping to organise the Biochemical Society focused meeting ‘The Keap1/Nrf2 pathway in health and disease’ along with Dr Maria O’Connell (East Anglia) and Prof Paul C Evans (Sheffield) to be held in Robinson College, Cambridge in late 2014-early 2015.