Several small and quite different ME/CFS studies have come up with the same tantalising finding – and now a team of two very talented resarchers, one a patient, are embarking on a robust replication that could help move the whole field forward.

The finding? That if you take blood from people with ME/CFS, and add it to healthy cells grown in the lab, it changes the cells’ behaviour, while blood from healthy people does not.

This could be a major biological clue to what is going wrong in ME/CFS, if the results from the early studies – all small and most unpublished – hold up. Confirming the finding would be a major boost to efforts to understand the mechanism of the disease and could lead to a diagnostic test.

Enter Dr Audrey Ryback and Charlie Hillier, her partner both in life and science. Audrey gained her PhD last year on a prestigious Wellcome programme, won funding from Action for ME, and is now working with Professor Chris Ponting at Edinburgh University.

Charlie, unable to take up a university place to do medicine because he developed ME/CFS, later did a molecular biology degree when his illness improved. Audrey and Charlie met in their first term at university.

Charlie had a year in industry with the prestigious Sanger Institute and was the lead author on a paper on cell biology in malaria. He gained a place to do a PhD at the European Molecular Biology Laboratory, as impressive as it sounds, but had to drop out after a bad relapse, and he hasn’t been able to work formally since. I interviewed the two of them together for this blog.

The first to observe this ‘something in the blood’ effect were Norwegians in 2016, led by Dr Oystein Fluge and Professor Olaf Mella, who ran the rituximab trial for ME/CFS, and by Professor Karl Tronstadt. Dr Ron Davis also found an effect, using his novel ‘nanoneedle’ device, as did researchers at three other independent research centres.

Why focus on ‘something in the blood’?

Blood has two main components: relatively large stuff like red and white blood cells, and the liquid in which they float, which contains many soluble factors such as antibodies. And all five labs that looked have found that it is this liquid (presumably, the molecules in it) from people with ME/CFS that appears to change how healthy cells behave.

Audrey and Charlie see these findings as the best biological lead for understanding ME/CFS. Their first goal is to replicate the Norwegian study, which found that serum (the liquid left over after blood has clotted) affects the ability of muscle cells grown in the lab to produce energy.

They chose this study out of the five that found an effect because:

• It uses proven, commercially available technology so they can be sure they can replicate the original method.
• The results are straightforward to interpret as they show exactly what part of energy metabolism changes. That’s very different from, say, the results for the nanoneedle, which are hard to interpret.
• The technology used in the study can process large numbers of samples.

The replication study

The original study used a small sample of 12 patients and 12 controls. To get more confidence in their results, Audrey and Charlie have upped that to 67 patients and 53 controls, significantly more than are used in most ME/CFS laboratory studies.

Their new study selected people using the DecodeME protocol. This requires people to confirm they have a medical diagnosis and also meet either the Institute of Medicine or Canadian Consensus criteria, based on DecodeME’s symptom questionnaire.

For the main study, muscle cells grown in the lab, called myoblasts, will be incubated in serum from patients or controls. Then Audrey and Charlie will use a Seahorse Analyzer – a piece of kit that measures energy metabolism in real time – to test the ability of myoblasts to produce energy in two ways:

• By glycolysis, in which glucose is metabolised without using oxygen to produce small amounts of ATP, the cell’s go-to energy molecule. This process can produce acid. 
• In oxidative phosphorylation, in which oxygen is consumed and mitochondria produce far more ATP.

This will show whether the findings of the Norwegian study replicate – that is, that there is something in the serum from people with ME that affects how mitochondria work.

Painting cells

Audrey and Charlie will also look at the effect of serum using cell painting. This technique uses fluorescent dyes to ‘paint’ key organelles of the cell (structures such as the mitochondria, nucleus and endoplasmic reticulum). Any changes in the appearance of the organelles revealed by the painting can give clues to how molecules in the serum are affecting cell biology.

Cell painting will also allow Audrey and Charlie to look for the mitochondrial fragmentation seen by Dr Bhupesh Prusty in the presence of serum from people with ME/CFS.

If either of these approaches – the Seahorse Analyzer or the cell painting – is a success, that itself would be a big achievement. It should also pave the way to better understand ME/CFS by helping shape testable ideas about what is going wrong. It could even lead to a diagnostic test.

What happens if they strike gold?

Audrey and Charlie would also want – if they have the time and funds – to narrow down what it is that affects cells. This is a needle-in-a-haystack problem, as serum contains many different molecules.

The team would probably start by splitting serum into fractions by size, from small to large molecules. The smallest would contain lipids and small molecules, while the largest would contain antibodies and protein complexes, and if one fraction contained the active component, it could be split further. Audrey and Charlie might also process serum biochemically, for example, by removing all the proteins and seeing if the serum still affected cells. They have enough processed blood samples to enable a lot of testing.

Even if this work doesn’t identify specific molecules, it should dramatically narrow down the possibilities, perhaps to a type of molecule, such as antibodies

And what if the original findings don’t replicate?

Publishing a finding that the study result didn’t replicate would be helpful too. ME/CFS research is awash with hundreds of interesting findings that have never been followed up. Most of them are probably wrong (see Why most published findings are false). Replications like Audrey and Charlie’s can show other researchers which findings are important and which are not – and that will help speed up research.

When will we see results?

Audrey only took up her position in September. By the end of February this year, she and Charlie had collected all the samples, with help from Caroline Dalton at Sheffield Hallam University (and Audrey stressed, from people with ME who shared posts on social media). They hope to have the main experiments completed later this year and publish their results next year.

This is fast going, and exciting stuff. It was a huge pleasure to talk to Audrey and Charlie, who have a clear vision of how to make progress and are obviously committed to patient involvement in studies. ME/CFS research has gained two brilliant minds, at least for now.

Coming next: Do young scientists have any future in ME/CFS research?

‘Ring-fencing from the big funders is essential if we are to get anywhere with ME research.’ Audrey Ryback

People like Audrey and Charlie are the future of ME/CFS research. Audrey’s funding runs out next year, yet there is next to no funding for ME/CFS research, which is often shunned by the big funders. We all lose out if people like her are forced to go elsewhere. What can be done?

Image credit Blood Image by OpenClipart-Vectors from Pixabay