Assessing cellular energy dysfunction in CFS/ME using a commercially available laboratory test, 2019, Morten, Newton et al

Sly Saint

Sly Saint

Senior Member (Voting Rights)
Abstract

The mitochondrial energy score (MES) protocol, developed by the Myhill group, is marketed as a diagnostic test for chronic fatigue syndrome/Myalgic Encephalomyelitis (CFS/ME). This study assessed the reliability and reproducibility of the test, currently provided by private clinics, to assess its potential to be developed as an NHS accredited laboratory test. We replicated the MES protocol using neutrophils and peripheral blood mononuclear cells (PBMCs) from CFS/ME patients (10) and healthy controls (13). The protocol was then repeated in PBMCs and neutrophils from healthy controls to investigate the effect of delayed sample processing time used by the Myhill group. Experiments using the established protocol showed no differences between CFS/ME patients and healthy controls in any of the components of the MES (p ≥ 0.059). Delaying blood sample processing by 24 hours (well within the 72 hour time frame quoted by the Myhill group) significantly altered many of the parameters used to calculate the MES in both neutrophils and PBMCs. The MES test does not have the reliability and reproducibility required of a diagnostic test and therefore should not currently be offered as a diagnostic test for CFS/ME. The differences observed by the Myhill group may be down to differences in sample processing time between cohorts.
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If I've understood........Dr Myhills test doesn't work.

full paper here
https://www.nature.com/articles/s41598-019-47966-z
 
Yeah, looks that way.

I thought the below comment was quite vocal.

"After a diagnosis of CFS/ME is made using the MES test, patients are subsequently sold supplements in order to treat their CFS/ME, despite there being no placebo-controlled trial to show their effectiveness. The first peer-reviewed publication regarding the MES test from the Myhill group came after they had already been using the test and supplement regime with CFS/ME patients despite there being no published evidence of the effectiveness, reliability, or reproducibility of the test."
 
jamari said:
Yeah, looks that way.

I thought the below comment was quite vocal.

"After a diagnosis of CFS/ME is made using the MES test, patients are subsequently sold supplements in order to treat their CFS/ME, despite there being no placebo-controlled trial to show their effectiveness. The first peer-reviewed publication regarding the MES test from the Myhill group came after they had already been using the test and supplement regime with CFS/ME patients despite there being no published evidence of the effectiveness, reliability, or reproducibility of the test."
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Exactly. She's making a killing down at Myhill farm. The tests provided are extremely expensive, and the results looked so convinving in the paper she published. it seems this was all due to sampling time and storage?
 
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I hadn't been quite aware how un-evidenced her test and treatments are.
The Myhill group have recently altered their protocol to use PBMCs instead of neutrophils, however, this research has not been published and we have no information on the control ranges used and whether they were developed from blood samples processed over 24 hrs. The CFS/ME PBMC study by Tomas et al. did show the utility of using PBMC using the Seahorse extracellular flux analyser to study energetics3. However, the sudden switch to using PBMCs for the MES protocol appears to be as a result of criticism over the use of neutrophils rather than being an evidence lead change. There has been no data published from the Myhill group regarding the suitability of PBMCs with their previously established protocol, or any publication of results with the new cell type. After a diagnosis of CFS/ME is made using the MES test, patients are subsequently sold supplements in order to treat their CFS/ME, despite there being no placebo-controlled trial to show their effectiveness. The first peer-reviewed publication regarding the MES test from the Myhill group came after they had already been using the test and supplement regime with CFS/ME patients despite there being no published evidenced of the effectiveness, reliability, or reproducibility of the test. Additionally, the MES test has not been conducted using other patient groups with fatigue as a core symptom, therefore its specificity for CFS/ME has not been confirmed.

If energetic dysfunction is to be used as a marker of CFS/ME its exact role in the condition needs to be better understood including studying energetic dysfunction in other fatigue groups. Only when we have clearer understanding of the disease process and knowledge of specific factors shown to be different in CFS/ME should we consider developing a diagnostic test to aid in treatment strategies and determining outcome in clinical trials.

Clinicians approached by patients with results from the MES test should be advised to interpret the results with caution, while patients considering paying for the test should be advised of the lack of supporting scientific evidence. The test in its current form does not have the reliability or reproducibility required of a diagnostic test and therefore should not be offered by the NHS or private clinics as a diagnostic test for CFS/ME. Other tests of energetic dysfunction could be developed using the seahorse extracellular flux assay but more research is required as to the meaning of the results in the aetiology of CFS/ME before a test using this approach should be developed.
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InitialConditions said:
this is the original paper: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2680051/

Could problems with storage and sample time really account for the original results, which show clear demarcation between HC and PWME, and also strong correlation between degree of disfunction and illness severity?
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If I am reading the new paper correctly, the thing they claim to be measuring is almost unmeasurable given variation in sample times as may be found by patients samples at poorly controlled temperatures being shipped to them.
In addition, there are severe questions about what they are in fact measuring.

And there is not enough data in the original paper to rule out storage delays between HC and PWME being the whole of the difference.
 
Jim001 said:
Is it unusual for a failed replication study, in ME/CFS no less, to appear in Nature?
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They do publish failed replication studies, not as often as you might like - but you can't make statistics from single events.

I would truly like to know the editorial process and reviewers on this, and hope no part of the decision was to bash a biomedical approach.
 
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This strikes me as a big step forward for the field.

I remember reading the original paper and feeling that the results looked almost too good to be true. But the acid test is always a replication attempt, ideally by an independent group and that’s exactly what’s happened here.

An independent group is more likely to be alive to any issues of implementation, such as different processing times and temperatures between patients and controls, which appears to be the issue here.

Thanks to the ME Association for funding this work.

To be fair to the original authors, we should wait to see their response to this new paper. But it does very much look like their test is not reliable and so should no longer be sold to ME/CFS patients.
 
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I agree. This is the sort of study we should be encouraging. It shows that there are people in biomedical ME research committed to getting reliable answers.

I found the original Myhill paper very hard to make sense of. It looked like a clear cut result but it was very unclear to me what it was measuring or why a result like that should explain symptoms. One of the problems of study peripheral blood cells is that by and large they are not doing anything. They are in transit to places where they might be active. In a disease situation the turnover is likely to change so interpretation is very hard. In all the disease I have worked on where white blood cells are heavily involved in tissue pathology you find nothing much in blood cells.
 
According to the PACE authors and their acolytes, since this trial did not show results that we should like (according to their model and claims of the past several decades anyway), we should be angry about it and harassing the researchers over it since they had the promise of showing results we should like and didn't.

Strange how that works out. They claim we are anti-science. I claim they are just bad at it and use it as a cover for promoting a belief system with zero consideration for the impact of their work in real life. Big difference.

Objective evidence or bust, whatever it shows. This is how science works. No moving goalposts, no hyping, no displeasure at null results, no fraud to turn negative results into claims of results so positive they amount to a complete cure if you believe in the treatment. Observe, hypothesize, falsify (or fail to). That's how it works and that's what we want.
 
I am glad this research was done although I share some confusion

I had this test many many years ago. It’s a shame it wasnt validated until now and probably a lot of patients money wasted. My results showed low ATP which now they’re saying was not reliable. I see dr M as arrogant on the fatigue issue but equally she was stabbing in the dark when there was no research to give anyone any other treatments and patients wanted to try anything . It was a reasonable hypothesis, we are still looking into atp today. Unfortunately her simple ideas for supplements didn’t fix the problems we are still trying to understand.
 
It seems Norman Booth, had similar concerns about the state of the Neutrophils which he discussed with John Mclaren Howard the one who came up with the test.
Source : https://forums.phoenixrising.me/threads/mitochondrial-and-energy-metabolism-dysfunction-in-me-cfs-—-myhill-booth-and-mclaren-howard-papers.47488/post-785000
A number of scientists have quoted published results of other studies of neutrophils that appear to disagree with John’s measurements. I have worried about this and have had several discussions with John about the disagreements. I am convinced of John’s integrity and talent. He measures neutrophils when they are still alive and are in a sterilized, aerated environment. Here they are going about their task of sensing the presence of foreign bodies and pathogens using their motility and chemotaxis. They are not in the process of honing in on pathogens or carrying out phagocytosis, which heavily rely on glycolysis. In one of the first steps in the ATP Profile, John inhibits mitochondrial function with sodium azide, but for only 3 minutes. This short period is adequate and also short enough to prevent apoptosis of any of the neutrophils. In some other studies neutrophils have been inhibited with the toxin Rotenone for as long as 6 hours [4]. If any of the cells are still alive they will have to have found some other pathway to provide them with ATP. As of 1st February 2016 John has made a change in his procedure, and now measures mixed leukocytes containing monocytes as well as neutrophils. There are only small differences from neutrophils only. John is not happy to use lymphocytes alone because of the varying subtypes present and the responses to infection.
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It does seem today's paper uses sodium azide for 3 minutes which others studies trying to replicate did not.
ADP to ATP efficiency
The efficiency of the conversion of ADP to ATP was monitored in the presence of excess magnesium with and without the complex IV inhibitor sodium azide. ATP concentration was determined as described previously. Following treatment with the inhibitor the cell suspension solution was centrifuged to remove the inhibitor and the pellet resuspended in SBS (combination of NaCl and KCl phosphate buffered to pH 7.8). This solution was left at room temperature for 3?minutes before being centrifuged again and the supernatant removed.
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Norman Booth was big on trying to get the work replicated to prove or disprove the test, and also bringing in Karl Morten. These are his own words.
But there is no better course than to have some other group to repeat what John has done or come up with some other method to confirm or disprove his results. Fortunately, the Ramsay Research Fund of the ME Association is now involved in funding four research projects involving mitochondrial function, including a new project led by Dr Karl Morten and Professor Joanna Poulton at the University of Oxford [5].
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Thank you Norman, rest in peace.
 
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The authors write in the Acknowledgements section
We would like to acknowledge Dr Norman Booth who sadly died last year. Norman has been instrumental in bringing the Oxford group into the CFS/ME research domain. Although Norman will be disappointed by our findings he would have appreciated the need to search for a robust diagnostic test for CFS/ME. This will be hopefully come with more straight forward approaches than assays involving live cells which are notoriously difficult to develop into diagnostic tests. We would also like to thank the ME Association, the Medical Research Council, and Action for ME.
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