(reproduced with permission from the combined notes of Duncan Cox and Jacqui
“OPEN MEDICAL MEETING”: LEAMINGTON SPA, 3 rd SEPTEMBER
Presentation by Dr John Gow
The first part of this afternoon organised by the ME Association was an excellent presentation by Dr John Gow, Senior Lecturer in Clinical Neuroscience at the University of Glasgow . Dr Gow, who has a background in genetics, neurology and virology, spoke about his ground breaking gene expression research and afterwards he answered questions on this work.
The aim of the study was to determine any characteristic changes in the biochemical pathways of patients with ME/CFS using DNA Chip microarray analysis (Affymetrix gold standard) of the entire human genome (33 000 genes ) construct a gene expression map of ME/CFS. It would also provide a rational basis for targeting pharmacological interventions.
Eight male ME/CFS patients, 18-54 years (mean 36yrs) were selected for the initial study from the Glasgow Southern General hospital using the 1994 CDC (Fukuda) criteria - they were compared them with seven healthy matched controls, aged 22-58 years (mean 34 yrs). Dr Gow said that the patients selected were considered to be "ding dong cases of ME” - to coin a Professor Peter Behan phrase. Dr Gow stated that up to 75% (sic) of patients that had been under an “ME label” at the hospital had been re-diagnosed with other conditions, such as MS, Lupus etc.
Blood samples were taken from the participants from which the RNA was isolated from peripheral blood mononuclear cells and analysed using a technique known as DNA chip microarray analysis. The sample “chips” were then scanned by computer to identify which of the 33,000 genes were either ‘up' or ‘down' regulated in comparison to the control (ie whether a gene was switched ‘off' when the control was ‘on' or vice versa). The work is at a sub-nanotechnology level with each patient's RNA sample producing 300 pages of data. It is also expensive as the chips are £1,000 each (one chip is needed for each patient or control sample). Therefore the very small sample was used initially to get the full gene picture, then other less expensive techniques could be used to test and confirm the specific gene patterns found, the microarray analysis having shown which genes to target. A total of 50 patients have now been studied.
It was interesting to see a slide comparing the different genetic studies that are currently taking place and the number of genes that each study has analysed:
Vernon ( Atlanta ) - 1,716 genes
Kaushik 1 ( London , Chronic Fatigue Syndrome Research Foundation) - 9,522 genes
Gow/Chaudhuri ( Glasgow ) - 33,000 genes (which actually amounted to 40,000)
The same gene signature was found in all samples. The study found eight "putative bio-markers" by the microarray assay and they were subsequently confirmed by using the less expensive “RT-PCR” and “western blot analysis” techniques with a larger sample. Separate lists were obtained for upregulated and downregulated genes, the most significant of which were up to 4 times more active than in controls. Twelve of the genes identified as being either up or down regulated were in the “oxygen transport group”. The top ones were Haemoglobin gamma A and Defensin alpha 1, Thrombospondin 1, Chondroitin Sulphate, Dynamin 1, Phosphatidyl Serine, Tubulin ß, Arginase. Prostaglandin synthesis and Cox 1 & 2 were found to be up-regulated and the MHC Class I system down-regulated. There was phosphatidyl serine binding. T-cell receptors were switched off.
In conclusion, the gene signature indicates:
.• A shift of immune response with preferential antigen presentation to MHC Class II (up-regulation of MHC Class II receptors and down-regulation of the MHC Class I system) response, including suppression of NK cell receptors
• Oxidative Stress
• Increased Neutrophil Apoptosis (Early cell death)
These were the 3 main areas highlighted for clinical/therapeutic intervention – some of which appear to correlate with MERGE studies at the University of Dundee 2, 3
Dr Gow indicated there already exist useful interventions for these areas but he could not make specific recommendations until phases of trials were completed. However, he said if you gave that list to your doctor, he/she would most likely come up with the same sort of interventions as he had in mind.
Various other comments made included:
There were immune problems, but not auto-immune, although one possibility is an auto-immune reaction to entero-virus infection.
There is no evidence of entero-virus activity, but enteroviral sequences found. A virus is opportunistic rather than the cause, ie a virus takes advantage of the immune problem.
A wide range of infections can trigger the response but it is not caused by persistent entero-viral infection. However, the gene signature is indicative of some sort of infection – the immune system doesn't switch off. Possibly there is some as yet unknown virus.
The same genes are switched on in borrelia infection.
The symptoms of ME/CFS are similar to those of basic hypothalamic dysfunction.
The results had been submitted to "Nature" magazine but they have not yet been accepted for publication.
Dr Gow has been struggling to finance this research and in May 2005 the press reported the Scottish Enterprise Council had withdrawn its support for the study. It has only been through the generous donations from the likes of the MEA (27K) and MERGE (8K) plus a number of smaller private donations that have helped keep this project alive. Dr Gow said that as yet he hasn't applied to Medical Research Council for funding, but he hoped that a joint funding proposal with Professors Chaudhuri & Findlay from the National ME Centre in Romford would be forthcoming: he didn't give any indication of when, although he gave the impression that it might be reasonably soon.
Dr Gow is currently in dialogue with an American University ( Atlanta ) that is also carrying out gene research and he is also in contact with four centres in Europe , including a joint project in Amsterdam . Also Dr Ben Natelson in New Jersey is taking this work one step further into proteins, using proteomics.
Further testing of large samples and comparisons not only with healthy controls but also with other disorders are now necessary to confirm this gene signature as the signature for ME/CFS, but Dr Gow seemed quietly confident that this was the long-awaited breakthrough in identifying significant bio-markers for ME/CFS.
It is hoped that by identifying the significant putative bio-markers and the immune response category, existing drugs can then be selected and tailored to help in the management of ME/CFS. Beyond that this work may give pointers to pharmaceutical companies to develop new and effective drugs.
Currently this sort of testing is only possible in highly specialist labs, and so is not available for general use. However it is hoped that in the medium term it should be possible to develop a specific test that could be conducted by a hospital pathology lab. Dr Gow thought it conceivable that a simple ‘dip stick colour change' kit could be developed for use in the GP's surgery.
1. N Kaushik, D Fear, S C M Richards, C R McDermott, E F Nuwaysir, P Kellam, T J Harrison , R J Wilkinson, D A J Tyrrell, S T Holgate and J R Kerr.
N. Gene expression in peripheral blood mononuclear cells from patients with chronic fatigue syndrome. Journal of Clinical Pathology. 2005; 58; 826-832.
2. Kennedy G, Spence VA , Underwood C, Belch JJF. Increased Neutrophil Apoptosis in Chronic Fatigue Syndrome. Journal of Clinical Pathology .2004; 57:891-3.
3. Gwen Kennedy, Vance Spence, Margaret McLaren, Alexander Hill, Christine Underwood & Jill JF Belch. Oxidative stress levels are raised in chronic fatigue syndrome and are associated with clinical symptoms. Free Radical Biology & Medicine 2005: 39(5): 584-9
Question & Answer Session
The second part of the meeting consisted of a question and answer session facilitated by Dr Charles Shepherd. The members of the medical panel were Dr John Gow, Dr Anne Macintyre, and Dr Rashmin Tamnhe, a paediatrician from Leicester . There were questions on a wide variety of topics including disability benefit assessment problems; the role of infection; and ME/CFS in children and adolescents. The session finally closed at 5pm but by that time many of the audience had left! My “key” notes were as follows:
• Dr Gow advised ME/CFS patients to steer well clear of the anti-viral drug Ampligen.
• Dr Anne Macintyre commented that many Examining Medical Practitioners (that she knew in the Bristol area) were of poor quality - which I guess we knew anyway! She also advised patients against resting before medicals, believing that it is better for the examining doctor to see you as you really are!
• Dr Charles Shepherd, said that the APPG have asked for a meeting with the Chairman of the Medical Research Council Professor Colin Blakemore about funding ME/CFS research in general
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