Dr. Paul Fernyhough
Division of Neurodegenerative Disorders
Cell Biology of Neurodegeneration Lab, Division of Neurodegenerative Disorders
Pharmacology & Therapeutics, University of Manitoba
Pharmacology and Therapeutics, Max Rady College of Medicine
Rady Faculty of Health Sciences, University of Manitoba
The WHO informs us that by 2025 there will be 300 million sufferers from diabetes worldwide – a figure approximately equal to the population of the USA. Neurobiologist Dr. Fernyhough is studying the etiology of the peripheral nerve damage observed in patients with diabetes. In addition he is researching the link between Alzheimer’s disease and Type 2 diabetes. “In patients with Alzheimer’s disease there is an increased risk of developing diabetes and these patients exhibit more severe and accelerated memory loss” says Dr. Fernyhough. Our studies are focused on identifying key signaling pathways that are impaired in animal models of Alzheimer’s disease. A major direction of the lab is to determine whether improper insulin signal transduction in neurons is central to axon and neuronal loss.
Abnormal peripheral nerve function in diabetic neuropathy
Evidence of neurodegenerative disease has been found in the peripheral nervous system in diabetes – commonly called diabetic neuropathies that involve damage to peripheral sensory neurons. Currently close to 50% of diabetic patients develop some form of peripheral nerve disease, which can lead to loss of protective sensation and limb amputation. The Aboriginal populations of Canada and the USA are experiencing an explosion in the incidence of type 2 diabetes. The incidence is expected to rise 10-fold in the next 10 years in First Nations peoples of Canada. Manitoba has one of the largest numbers of Aboriginal persons in Canada and so the health burden in this province is becoming severe. Diabetic sensory neuropathy and retinopathy are particularly severe complications in these populations. Approximate direct health costs in Manitoba for neuropathy (including amputation and foot treatment) are CA$100-150 million per annum. This excludes the social costs of loss of work, relocation and rehabilitation. The human cost is enormous. Young patients, 30 years of age, are undergoing amputations and death from infection is increasingly occurring.
Why is this work important?
Currently, there are no effective treatments for any of these serious neurological diseases. Dr. Fernyhough’s research is focused on identifying the key cellular/molecular pathways that are regulated by insulin in the maintenance of mitochondrial function and to determine what goes wrong in diabetic sensory neuropathy and Alzheimer’s disease.
What techniques and equipment are used in this laboratory?
- In vitro and in vivo models
- Animal models of type 1 and 2 diabetes (STZ rat and mouse; ZDF rat)
- Primary sensory neuron cell culture
- Molecular studies
- Confocal microscopy – inverted and upright
- Real time video microscopy – calcium, mitochondrial function, free radicals (standard light and confocal – Zeiss LSM 510)
- In vitro enzyme assays
- Viral-mediated transfection – lentivirus and adenovirus
- Western and Northern blotting (quantitative)
- Real time RT-PCR
- The Division of Neurodegenerative Disorders (DND) occupies 5,000 square feet of newly-renovated laboratory space. This includes; laboratories for six principal investigators (2 new PIs to be hired in the next year), three culture rooms, a major equipment room, a Carl Zeiss LSM510 confocal microscopy room, a room for calcium imaging, a Carl Zeiss Axioskop II light upright microscope suite, a Bio-Rad Fluor-S imaging suite, HPLC room, a dark room, a walk-in cold room, two non human surgery and behavioral monitoring rooms, and a conference/student room.
About Dr. Paul Fernyhough
Dr. Fernyhough was born and educated in East London, UK, and performed his B.Sc. degree in Biological Sciences at the University of Essex. Dr. Fernyhough performed his PhD in biochemistry in the department of Biochemistry (department of Sir Hans Krebs) at University of Sheffield in the UK. He also performed postdoctoral research at Colorado State University, Kings College London (department of Maurice Wilkins) and as a Wellcome Trust Postdoctoral Fellow at St Bartholomew’s Medical College (department of Sir John Vane). Drs Krebs, Wilkins and Vane are all Nobel Prize winners. All of these positions spanned 1985-1998. Dr. Fernyhough subsequently worked for 5½ years (1998-2004) as a fully tenured lecturer in the School of Biological Sciences (now the Faculty of Life Sciences) at the University of Manchester. Dr. Fernyhough’s general research interest is in the cell biology underlying neurodegenerative disorders of the peripheral and central nervous systems.
For more information, please contact:
Dr. Paul Fernyhough
Director – Division of Neurodegenerative Disorders at the St. Boniface Hospital Research Centre &
Professor, Dept. of Pharmacology & Therapeutics, University of Manitoba, Faculty of Medicine
Room R4046, 351 Tache Avenue
Winnipeg, Manitoba, Canada
Phone: (204) 235-3939
Fax: (204) 237-4092
Impaired calcium homeostasis and mitochondrial dysfunction in diabetic sensory neuropathy (CIHR-funded)
Image Approximately 50% of persons with diabetes present with sensory neuropathy which involves the dying back of distal axons and a failure of axons to regenerate. This leads to incapacitating pain, sensory loss and poor wound healing. The end result is lower extremity amputation (LEA) with approximately 8,000 diabetes-related amputations occurring each year in Canada and the expectation of a 5-fold increase over the next 10 years due to increased incidence of type 2 diabetes. Approximately $1.5 billion in Canada in 1998 was incurred for palliative treatment of neurological complications of diabetes, and at present there is no effective drug therapy. Diabetic sensory neuropathy and neurodegenerative diseases of the CNS disease, e.g. Alzheimer’s disease and ageing, share common pathological features involving the loss of distal axonal processes. Abnormal neuronal calcium (Ca2+) homeostasis has been implicated in numerous CNS diseases including diabetic sensory neuropathy. The endoplasmic reticulum (ER) regulates Ca2+ homeostasis and this process is linked to regulation of mitochondrial function and the activity of the anti-apoptotic transcription factor, NF-kB.
The major aim of this proposal is to determine the mechanisms whereby diabetes impairs ER-dependent Ca2+ homeostasis in dorsal root ganglia (DRG) sensory neurons. We will test the hypothesis that abnormalities in Ca2+ homeostasis in sensory neurons in diabetes are linked to impaired mitochondrial function and aberrant NF-kB signalling and lead to deficits in axon function that trigger distal axonal loss.
Mechanisms of impaired AKT signaling and associated mitochondrial dysfunction in sensory neurons in diabetic neuropathy (CIHR funded)
Diabetic sensory polyneuropathy is the most common form of peripheral neuropathy and has an increasingly detrimental impact on human health and associated costs – there is no treatment. The etiology remains poorly understood, however, a dying back of distal axons and a failure of axonal regeneration are key features of this neurodegenerative disease. Impairment of neuronal anaerobic or aerobic metabolism would be expected to have a significant deleterious effect on axonal function. Our studies demonstrate that mitochondria within the lumbar dorsal root ganglia (DRG) of streptozotocin (STZ)-diabetic rats (animal model of Type I diabetes) are depolarized. Our ongoing studies show that mitochondrial function and associated ATP levels in adult sensory neurons are regulated by insulin-dependent modulation of the phosphoinositide 3-kinase (PI 3-kinase)/Akt signaling pathway. Clearly in diabetes insulin and other neurotrophic growth factor-dependent support of neurons is sub-optimal. Therefore, the general purpose of this proposal is to identify the key cellular/molecular pathways regulated by insulin in the maintenance of mitochondrial function and to determine what goes wrong in diabetic sensory neuropathy.
The role of transcription factor, NF-κB, in the etiology of symmetrical sensory polyneuropathy in diabetes. (Cdn Diabetes Assoc. funded)
The development of symmetrical sensory polyneuropathy in persons with type 1 and 2 diabetes is a major cause of morbidity, leading to pain, limb ulceration, poor wound healing and in many cases lower extremity amputation (LEA). This crippling condition severely reduces the quality of life of patients and is an enormous financial drain of resources from Health Canada. There is no treatment, other than palliative foot care which reduces the severity of the disease. A key component of the pathology is the irreversible dying-back of distal axons with the longest myelinated and unmyelinated nerve fibers being preferentially targeted. Studies in animal models of diabetes reveal a heterogeneous etiology involving hyperglycemia and hypoinsulinemia which reduces peripheral nerve function through impairments in endothelial cell, Schwann cell and/or axon function and leading to generation of pain and neuropathy. Recent clinical work on human sural nerve biopsies has shown that early in the disease progression there is unmyelinated fiber loss and structural abnormalities in myelinating Schwann cells concomitant with microangiopathy in nerve blood vessels. However, these structural alterations were observed in tandem with a normal appearance in the axons of myelinated fibers. Hence, Schwann cells and the endothelial cells lining the endoneurial blood vessels have been proposed as primary targets of diabetes-related trauma in myelinated and unmyelinated nerve fibers. Recent work from our laboratories and by others has shown an elevation in activation of the transcription factor, NF-kB, in peripheral nerves of streptozotocin (STZ)-diabetic rats. NF-kB is activated under conditions of oxidative stress in a range of tissues and disease states and, in some cases, can contribute to cellular dysfunction. Consequently, we will test the hypothesis that activation of NF-kB in endothelial cells and Schwann cells of peripheral nerve fibers is aberrant in diabetic rats, and leads to impaired axon function, abnormal myelinated and unmyelinated fiber structure, reduced axonal regeneration and ultimately irreversible distal axon loss.
Growth factor-dependent regulation of neuronal metabolism. (funded thru National Sciences and Engineering Research Council of Canada).
The survival and growth of embryonic and adult neurons is regulated by neurotrophic factors, such as nerve growth factor (NGF) and insulin and insulin-like growth factors (IGFs). Neurotrophic factors control an array of signal transduction pathways that modulate the phenotype of neurons through adjustment of gene expression at the transcriptional, post-transcriptional and post-translational levels. For the neuron to perform such a range of catalytic and homeostatic operations there must be a constant supply of metabolites and energy, however, the processes whereby neurons regulate their own metabolism and energy production are poorly understood. We will test the hypothesis that neurotrophic factors enhance neuronal growth and survival through signal transduction pathways that directly control metabolite levels and ATP synthesis. Our preliminary studies in adult sensory neurons indicate that neurotrophic factors, such as NGF and insulin, enhance the mitochondrial function and raise ATP levels through activation of the phosphoinositide 3-kinase pathway (PI 3-kinase). Recent results indicate a central role for the multi-functional enzyme Akt, downstream of PI 3-kinase, in directing neurotrophic factor-dependent signals controlling neuronal metabolism. The objective of our future work will be to fully identify the pathways utilized by neuronal growth factors, focusing mainly on the PI 3-kinase/Akt axis, to modulate the full spectrum of mitochondrial bioenergetics in embryonic and adult neurons of the central and peripheral nervous systems (CNS and PNS).
Impact of drugs on axon sprouting and regeneration of adult sensory neurons.(funded thru JDRF-USA)
The current proposal plans to screen 1000 FDA approved drugs using an in vitro assay involving the culture of adult rat sensory neurons. The principal applicant has extensive experience with this culture system and over a 13 year period has used this methodology to study processes that regulate axon outgrowth, intracellular calcium levels, mitochondrial function, impact of high glucose and gene expression – all the studies being aimed at understanding the etiology of sensory loss in diabetes. This tried and tested culture system permits the in vitro assessment of axonal spouting and/or regeneration from adult sensory neurons from the dorsal root ganglia. A range of studies have previously shown this in vitro system to be relevant with regard to in vivo processes governing axon regeneration and sprouting.
Integrated tissue imaging lab. (funded thru Canadian Foundation for Innovation)
Technical restrictions currently limit study of the cell biology of neurodegenerative and vascular disorders to cell cultures. The applicants aim to bring their research to the forefront on the global scene by studying real time events in cells that are situated in intact tissues. With the requested infrastructure, this goal will be achieved by allowing simultaneous assessment of several aspects of cell function in brain slice preparations, and the proposed facility will form an integrative live tissue imaging lab that will allow the applicants to raise their current research to a multi-disciplinary, internationally recognizable level. The following sections outline the infrastructure components and their projected uses. The applicants occupy four adjacent labs on the 4th floor of the St. Boniface Hospital Research Centre, and the equipment will be installed in a shared facility.
Fernyhough Top 10 publications – Nov 2012
Roy Chowdhury, S.K., Smith, D.R., Saleh, A., Schapansky, J., Marquez, A., Gomes, S., Akude, E., Morrow, D., Calcutt, N.A. and P. Fernyhough (2012). Impaired AMP-activated protein kinase signaling in dorsal root ganglia neurons is linked to mitochondrial dysfunction and peripheral neuropathy in diabetes. Brain. 135, 1751-1766. (if, 9.23)
Saleh, A., Roy Chowdhury, S.K., Smith, D. R., Balakrishnan, S., Tessler, L., Martens, C., Morrow, D., Frizzi, K., Calcutt, N.A. and P. Fernyhough (2012). Ciliary neurotrophic factor activates NF-B to enhance mitochondrial bioenergetics and prevent neuropathy in sensory neurons of streptozotocin-induced diabetic rodents. Neuropharmacology. In Press. (if, 4.81)
Roy Chowdhury, S.K., Smith, D.R. and P. Fernyhough (2012). The role of aberrant mitochondrial bioenergetics in diabetic neuropathy. Neurobiology of Disease. In Press. (if, 5.12)
Akude, E., Zherebitskaya, E., Roy Chowdhury, S.K., Smith, D.R., Dobrowsky, R.T. and P. Fernyhough (2011). Diminished superoxide generation is associated with respiratory chain dysfunction and changes in the mitochondrial proteome of sensory neurons from diabetic rats. Diabetes. 60, 287-299. (if, 8.9)
Jackson, A.C., Kammouni, W., Zherebitskaya, E. and P. Fernyhough (2010). Role of oxidative stress in rabies virus infection of adult mouse dorsal root ganglia neurons. Journal of Virology. 84, 4697-4705. (if, 5.31)
Roy Chowdhury, S. K., Zherebitskaya, E., Smith, D.R., Akude, E., Chattopadhyay, S., Jolivalt, C.G., Calcutt, N.A. and P. Fernyhough (2010). Mitochondrial respiratory chain dysfunction in lumbar dorsal root ganglia of streptozotocin-induced diabetic rats and its correction by insulin treatment. Diabetes. 59, 1082-1091. (if, 8.9)
Zherebitskaya, E., Akude, E., Smith, D.R. and P. Fernyhough (2009). Development of selective axonopathy in adult sensory neurons isolated from diabetic rats: role of glucose-induced oxidative stress. Diabetes. 58, 1356-1364. (if, 8.5)
Fernyhough, P., Smith, D.R., Schapansky, J., Van Der Ploeg, R., Gardiner, N.J., Tweed, C.W., Kontos, A., Freeman, L., Purves-Tyson, T.D. and G.W. Glazner (2005). Activation of NF-κB via endogenous TNFregulates survival of axotomized adult sensory neurons. Journal of Neuroscience. 25,1682-1690. (if, 8.3)
Middlemas, A., Delcroix, J.-D., Sayers, N.M., Tomlinson, D.R. and P. Fernyhough (2003). Enhanced activation of axonally transported stress-activated protein kinases in peripheral nerve in diabetic neuropathy is prevented by neurotrophin-3. Brain. 126, 1671-1682. (if, 8.0)
Huang, T.-J., Price, S., Chilton, L., Calcutt, N.A., Tomlinson, D.R., Verkhratsky, A. and P. Fernyhough (2003). Insulin prevents depolarization of the mitochondrial inner membrane in sensory neurons of Type I diabetic rats in the presence of sustained hyperglycemia. Diabetes. 52, 2129-2136.
2008 Juvenile Diabetes Research Foundation, Mary Jane Kugel Award
2007 University of Manitoba Presidential Outreach Award
2006 Juvenile Diabetes Research Foundation, Mary Jane Kugel Award
1993 – 1998 Wellcome Trust Postdoctoral Fellowship Pharmacology $ 700,000
Department of Pharmacology
Queen Mary & Westfield College
University of London, United Kingdom
1991 – 1993 Merck, Sharp & Dome Pharmacology $100,000
Postdoctoral Fellowship Research Award
St. Bartholomew’s Medical College
Queen Mary & Westfield College
University of London, United Kingdom
1987 – 1989 MRC Postdoctoral Fellowship Award Cell Biology $ 90,000
MRC Cell Biophysics Unit, King’s College
London, United Kingdom
1981 – 1984 BBSRC PhD Scholarship Research Award Biochemistry $ 30,000
University of Sheffield, United Kingdom
Current Research group:
Dr. Ali Saleh – research associate (CIHR)
Dr. Subir Chowdhury – research associate (JDRF)
Dr. Wafa Kammouni – research associate (CIHR)
Mr. Randy Van Der Ploeg – technician (JDRF)
Dr. Darrell Smith – research associate (SBRC)
Ms. Lori Dunn – technician (JDRF)
Mr. Dwane Morrow – technician (JDRF)
Mr. Tarek Habash – MSc student
Ms. Mahalakshmi Razdan – MSc student
Zaahra Waly – PhD student
Listing of active grants/PER ANNUM
Juvenile Diabetes Research Foundation (JDRF # 1-2011-590)
Total US$495,000. Operating grant: Metabolic regulation of neuronal mitochondrial function in diabetes. PI: Fernyhough, P.; Co-PI: Czubyrt, M., Univ of Manitoba.
Canadian Institutes of Health Research (CIHR # RPA 113765)
Total $200,000. Operating grant: Targeting the cytokine/NF-B axis in diabetic neuropathy. PI: Glazner, G.W.; Co-PI: Fernyhough, P.
Juvenile Diabetes Research Foundation (JDRF # 17-2011-252)
Total $605,273. Operating grant: Muscarinic receptor antagonists for treatment of diabetic neuropathy. PI: Fernyhough, P.; Co-PIs: Calcutt, N.A., UCSD and Kotra, L., Univ of Toronto.
Juvenile Diabetes Research Foundation (JDRF # 17-2010-795)
Total $309,925. Operating grant: DLK/MLK inhibitors for the treatment of diabetic neuropathy. PI: Dewhurst, S., Univ. of Rochester; Co-PIs: Fernyhough, P., Calcutt, N.A., UCSD, and Goodfellow, V.S., Califia Bio, Inc..
Juvenile Diabetes Research Foundation (JDRF # 39-2009-647)
Total $596,423. Operating grant: Diabetic neuropathy, neuronal insulin and its interaction with AGE-RAGE. PI: Zochodne, D., Univ of Calgary; Co-PIs: Fernyhough, P. and Toth, C., Univ of Calgary.
Juvenile Diabetes Research Foundation (JDRF # 17-2009-722)
Total $137,500. Operating grant: High content screening of sensory neurons. PI: Fernyhough, P.
Canadian Foundation for Innovation (CFI)
Total $978, 582. Leaders Opportunity Fund: High Throughput Bioimaging Facility. PI: Fernyhough, P.; Co-PI: Zahradka, P.
Canadian Institutes of Health Research (CIHR)
Total $200,000. Operating grant: Rabies virus-induced injury to neuronal processes: role of oxidative stress. PI: Jackson, A.; Co-PI: Fernyhough, P.
2007 – 2012
Canadian Institutes of Health Research (CIHR; # MOP-84214)
Total $600,000. Operating grant: Impaired calcium homeostasis and mitochondrial dysfunction in diabetic sensory neuropathy. PI: Fernyhough, P.; Co-PI: Glazner, G.W., Univ of Manitoba.
Canada Foundation for Innovation (CFI)
Total $119,000. Infrastructure Operating Fund: Integrated tissue imaging lab. PI: Anderson, C.; Co-PI’s: Anderson, H., Fernyhough, P., Albensi, B.
2006 – 2011
Canadian Foundation for Innovation (CFI)
Total $1,004,835. New Opportunities Fund: Integrated Tissue Imaging Lab Co-PI’s: Anderson, C., Fernyhough, P., Albensi, B., Anderson, H.
2006 – 2011
Natural Sciences and Engineering Research Council (NSERC; # 3311686-06)
Total $180,000. Individual Discovery Grant: Growth factor-dependent regulation of neuronal metabolism. PI: Fernyhough, P.
Roy Chowdhury, Subir