1 Webinar with Christopher Armstrong Christopher Armstrong2016 Webinar Series | Thursday, October 20, 2016 | 1:00 PM Eastern Webinar with Christopher Armstrong Christopher Armstrong University of Melbourne
2 About Our Webinars Welcome to the 2016 webinar series!The audience is muted; use the question box to send us questions Webinars are recorded, and the recording is made available on our YouTube channel: The Solve ME/CFS Initiative is a research organization and does not provide medical advice
3 Save the Dates! Thursday, November 10: Anthony Komaroff, MDAnthony Komaroff, MD, Simcox-Clifford-Higby professor of medicine at Harvard Medical School and senior physician at Brigham and Women’s Hospital in Boston, Massachusetts Thursday, December 15: Zaher Nahle, PhD, MPA Zaher Nahle, PhD, MPA , Vice President for research and scientific programs at Solve ME/CFS Initiative
4 Webinar with Christopher Armstrong Christopher Armstrong2016 Webinar Series | Thursday, October 20, 2016 | 1:00 PM Eastern Webinar with Christopher Armstrong Christopher Armstrong University of Melbourne
5 Melbourne Group A/Prof Paul Gooley University of Melbourne Protein Biochemist Expertise in NMR Dr Neil McGregor Once editor of Journal of Chronic Fatigue Syndrome Researching ME/CFS for 20+ years Expertise in metabolism Dr Henry Butt Formed Bioscreen Medical company that screens fecal samples for intestinal dysbiosis Researching ME/CFS for 20+ years Dr Don Lewis Formed CFS Discovery clinic Specialist clinic for managing and treating people with ME/CFS Treating people with ME/CFS for 20+ years
6 Our metabolomics work in ME/CFSYet to be published
7 Recent metabolomics studies and other associated studies
8 What is metabolomics? Metabolites are any small organic molecule or chemical (amino acids, sugars, drugs, etc.) Metabolomics is the study of metabolites External environment Alter Metabolites Alter Instructions to make Alter Initiate Proteins Genes Genes and the extension of genes (proteins) evolved to move around metabolites like scientists in a lab *Images extracted from lecture by David Wishart from University of Alberta Informatics and Statistics for Metabolomics workshop hosted by the Canadian Bioinformatics Workshops
9 Value of metabolomics to ME/CFSStress on a biological system produces symptoms that vary upon the individual. May cause variations between people with ME/CFS . ME/CFS still may be an umbrella term for a number of similar disorders (spectrum disorder). ME/CFS is diagnosed by symptoms. Metabolites are closely linked to symptoms. Developed metabolite tests can be used in clinical studies to test effectiveness of treatments. Metabolomics produce a large picture from which we are able to look for potential cause.
10 History of metabolomics in ME/CFSWent through hundreds of papers to find metabolite studies Summarised some studies in tables (pictured) Comparing multiple metabolomics studies: Oxidative stress environment Sub-maximal usage of mitochondria for energy Decrease of amino acids Issues with nitrogen metabolism (urea cycle)
11 Our metabolomics work in ME/CFS
12 Blood and Urine Metabolites
13 Creatinine normalisationUrine metabolites when corrected by creatinine. It may be possible that there is an extreme reduction in metabolite excretion. Need to do 24 hour urine collection. As glycolysis was inhibited we suggest it’s likely that creatinine is increased to compensate.
14 AST Energy without oxygen Glycolysis inhibitionCreatinine replacement? Too much oxygen/reperfusion Hypoxanthine to allantoin Creates reactive oxygen species Energy production with oxygen Reduced acetate -> Reduced lipolysis Amino acids used for energy Aspartate Transaminase (AST) AST Increase Decrease
15 Increased Aspartate TransaminaseAspartate Transaminase (AST) increased 3-fold. Top enzymes involved in using amino acids and lipids to fuel the TCA cycle. Enzymes of electron transport were reduced.
16 Hypometabolism ~30 metabolites in both the blood and urine of 59 women (34 ME/CFS vs 25 age-matched controls) 6 metabolites were significant altered in blood and 5 were decreased (83%) 5 metabolites were significantly altered in urine and 5 were decreased (100%) Comparable to Naviaux et al. that found 84% of altered metabolites in blood were decreased. In NMR we only saw water-soluble metabolites (no lipids). Advantage of NMR is that you can see all the atoms within a sample, we quantitated about 95%. Metabolites are made of carbon, hydrogen, oxygen, nitrogen. Total C, H, O in blood were equal between ME/CFS and controls. Total N was significantly decreased in blood of ME/CFS. Total C, H, O, N in urine were equal between ME/CFS and controls.
17 We’re all on a similar page!Metabolite studies of the past and most recent Increased oxidative stress Increased use of lipids for ATP Issues of purine metabolism Issue with folate cycle and methionine Reduced glycolysis/increase sugars Increased use of amino acids for ATP
18 What could it mean? Well these findings aren’t unique. This set of findings are actually similar to two acute disorders: - Starvation - Sepsis Differences between them and ME/CFS seems to align with both in some capacity. Similar mechanisms involved with ME/CFS but at a slow chronic pace. Provides clues.
19 Sepsis and Starvation Similarities to ME/CFS.Infection or stressor (both) Cytokine release (sepsis) Oxidative stress (both, more in sepsis) Mitochondrial dysfunction at oxidative phosphorylation site (both) Low blood volume (both) Hypometabolism (starvation) Hyperglycemia (starvation) Mitochondria use lipids and amino acids for ATP production Key difference to ME/CFS. Increased glycolysis (sepsis) Hypermetabolism (sepsis) Collagen breakdown - POTS, hypermobility, slow gastric motility?
20 Recovery? Yes people recover from sepsis and starvation.Monitored and drip fed because recovery has it’s own limiting factors. Refeeding syndrome. Food given to help starvation causes a sudden increase in metabolites in the blood, these metabolites can lower phosphate, potassium, magnesium, calcium, etc. Metabolism requires cofactors, minerals and vitamins to function. These deplete as metabolites deplete, reintroducing increases of metabolites can exhaust these even more. Our current studies: Longitudinal monitoring of people with ME/CFS to understand which combination of metabolites, cofactors, vitamins and minerals may have positive affects.
21 Our metabolomics work in ME/CFSYet to be published
22 Microbes in fecal samplesLive microbial count from fecal samples Clostridium spp. and Bacteroides spp. ferment digested compounds to SCFA Species of Clostridium have a wider variety of substrates for SCFA production Clostridium can make SCFA from peptides and amino acids Yet to be published
23 Metabolites in fecal samplesIncreased SCFA and isovalerate with decreased amino acids in fecal samples Production of fatty acids from proteins may create damaging byproducts in the gut Yet to be published
24 Metabolite correlationsAbnormal energy metabolites in blood positively correlated with amino acids in fecal samples of ME/CFS patients. No significant correlations of increased SCFA with metabolites in blood or urine in ME/CFS. Significant negative correlations of fecal SCFA with metabolites in blood involved in energy metabolism. Yet to be published
25 Metabolite correlationsShort chain fatty acids appear to cause hypometabolism event in controls. Studies show that SCFA have this effect by triggering AMPK. AMPK is the master switch that increases lipid and amino acid production of ATP. Expected the increase in SCFA to also indicate an effect on hypometabolism in ME/CFS. Maybe something more at work. Reduced amino acids in feces appears linked to altered energy metabolism in ME/CFS. Yet to be published
26 What could it mean? Gut ties in with both starvation and sepsis.Bacterial translocation and leaky gut -> inflammation. Bacterial translocation is normal, however altered bacteria may pose a threat. pH of the gut and blood may be very important here. During starvation, proteolysis of proteins occurs. First are proteins in the gut. Could this be responsible for slow gut motility or reduced dietary protein breakdown?
27 Limitations Metabolite studies have been on blood and urine so far.We can only make inferences on what is occurring in cells. Until we look at the metabolites in cells and mitochondria we can’t be sure of what is occurring. Metabolome is dynamic Your metabolome changes constantly and is individual. Taking a snapshot of dynamics in biology doesn’t tell you nearly enough. Patient vary and will show differences based on sample collection parameters. ME/CFS is dynamic Symptoms change over the day and over the course of the illness.
28 Current/Future StudiesLongitudinal metabolomics and genomics study on ME/CFS patients with monitored intervention. Metabolomics of ME/CFS cells. Production of large symptom survey database for potential patient stratification. Genetic markers and metabolite population studies.
29 Acknowledgements Questions…?
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32 Thank you for joining usThank you for joining us! Our mission To make ME/CFS understood, diagnosable, and treatable