Third Annual Community Symposium on the Molecular Basis of ME/CFS Sponsored by OMF - DISCUSSION

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FMMM1

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I think its a form of stalling. They present good ideas as excuses for not doing anything now. Divide and conquer or split the baby.
They have been using this excuse for a long time, in their request for submissions earlier this year i explained that this is silly, they are saying you need the chicken before the egg, if you want young researchers who have no reason to come into a dead field then you won't get them because they are saying we won't fund the incumbent players who could impart their experience and knowledge to the next generation plus with no funding now it does not look like there will be funding for anyone later. Empty promises don't pay the bills when they have been employing them for years.
A fake Mexican standoff
I think Vicky comes from a patient community background similar to this - a young family member ill with a severely disabling disease + she's a scientist (PhD). Plus she hasn't done this alone; I guess Walter Crozier has been a keen player in this turn around. Yes they do need to deliver the measures in the report --- they referred to this "not being just another report".

I generally share your cynicism about policy/politics stuff - I work in it (at a junior level) and I'm generally a cynic.

Ron Davis has long highlighted that unless there's funding researchers will move to other areas.
 

Alvin2

The good news is patients don't die the bad news..
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I think Vicky comes from a patient community background similar to this - a young family member ill with a severely disabling disease + she's a scientist (PhD). Plus she hasn't done this alone; I guess Walter Crozier has been a keen player in this turn around. Yes they do need to deliver the measures in the report --- they referred to this "not being just another report".
She has been one of the many spouting this fake Mexican standoff. Either she is deluded or she is not really on our side.
Even if her superiors are lying to her that means she is deluded by proxy (aka being used)

Ron Davis has long highlighted that unless there's funding researchers will move to other areas.
Until proven otherwise this is the NIH's real goal.
 

MonkeyMan

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Not having been able to follow the talks, however, reading the summaries here, I am concluding that work is progressing. However, it looks like there is still so much to do, and a very long way off for any treatment--decades. No one has cracked the illness yet. I just don't understand why. We can fly to Mars, to the Moon, build extraordinary computers, and yet we cannot figure out what is going on with this illness. It is just not comprehensible to me. It is also very alarming. Of course, this group is committed, and hard working and passionate--this is not at issue. But we are talking about a grave and seriously disabling illness. What is it that is amiss? I am heartbroken. I know several severely ill young people, rotting in bed, and declining daily. Lord have mercy.
@perrier, having battled ME/CFS for 35 years, I share your feelings of disappointment and sadness at the slowness, but I think to say any treatment is "decades" is away is exaggerating. There are many potential treatments being studied, and several of these are existing agents that would not require the lengthy FDA approval process. Even those that would may receive expedited approval. There is truly grounds for hope. For me, Jarred Younger's work is the most exciting and promising.
 

FMMM1

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@perrier, having battled ME/CFS for 35 years, I share your feelings of disappointment and sadness at the slowness, but I think to say any treatment is "decades" is away is exaggerating. There are many potential treatments being studied, and several of these are existing agents that would not require the lengthy FDA approval process. Even those that would may receive expedited approval. There is truly grounds for hope. For me, Jarred Younger's work is the most exciting and promising.
I agree with @perrier i.e. so much has been achieved in other areas - when there has been a political wish to do so.

A solution may come from another disease/research area. E.g. Cort Johnson has just published an article on a switch to focusing on neuro-inflammation in Alzheimer's disease [https://www.healthrising.org/blog/2019/09/16/alzheimers-chronic-fatigue-fibromyalgia/ ].

Some of the drugs they're looking at in Alzheimers, to reduce neuro-inflammation, are currently licensed for other diseases and in one case they are reformulating an existing drug i.e. to get it across the blood brain barrier. @MonkeyMan will be aware that Jarred Younger is a leading researcher in neuro-inflammation in ME.

Another thing we could try is lobbying (I do a little with ME Action):
Dr Vicky Whittemore (NIH) at Invest in ME Conference (2019):
"advocacy groups --- that's what makes the difference -
- when they [elected representatives] hear that, from people with the disease -
- advocates -- telling them [elected representatives] what's needed is really what makes the difference"
 

nandixon

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I was pretty disappointed not to hear about any progress on or even any mention at all(?) of the "something in the blood" finding at the Symposium. As a former research scientist it's very strange to me that the Stanford OMF group is not laser-focused on narrowing down what the component is in ME/CFS plasma that causes healthy cells to act like ME/CFS cells in the nanoneedle. Instead they seem to be going off in more and more different directions. Why go on more fishing expeditions when you've already located the fish and just need to reel it in?

The only thing I can figure is that it's the problem of the current iteration of the nanoneedle having a very low throughput (perhaps together with not having anyone who has the skillset to do natural substances isolation). And one would need a pretty high throughput in the course of gradually fractionating out and isolating and identifying the "something."

Figuring out what the something is could obviously lead to both a biomarker and more importantly a treatment. So I've been wondering if it might be possible to use the nanoneedle to raise money… for the nanoneedle.

@Janet Dafoe (Rose49), @Ben H

In particular, I'm wondering if it's been determined whether fresh blood is really necessary for the nanoneedle test, or if the plasma could be pre-processed in some way and frozen at some temperature ahead of time such that it could be sent in by patients from a distance away and then tested on healthy cells there at Stanford? There's seemingly no need for the whole blood.

I've seen some plasma processing techniques, for example, that allow for exosome analysis to be done at a later date after freezing and storage of plasma. (Note that I'm just using this as an example since I'm assuming it's not known whether an exosome is involved or not.)

Nanoneedle testing could then be offered as a clearly spelt out purely for fun, non-diagnostic test (i.e., lots of disclaimers) for a donation. (Not only patients but also someone they know who is healthy and doesn't have ME/CFS might send in plasma specimens for an extra donation.)

The money would then go towards making the nanoneedle more efficient, smaller, etc., and for determining what the something in the blood is.
 

raghav

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Is it possible to create tracer versions of Copaxone and SS-31 and see with which exosome they are interacting with or whether they are entering the sick cell and then interacting with something inside the cell and proceed from there ? Is such an experiment possible ?
 

ljimbo423

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Figuring out what the something is could obviously lead to both a biomarker and more importantly a treatment.
Also, if they find the something in the blood. Maybe they could trace it back to the root cause of ME/CFS. If they can do that, that could open the door for very effective treatments or possibly even ways to cure ME/CFS.
 
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@nandixon We have to also remember the researcher who worked on the nanoneedle has moved to a new position at UC Irvine so perhaps that has slowed down the work, even if he is still involved. It's taken 2 years to test 40 patients so it is clearly a slow process and the nanoneedle needs improved for throughput.

One concern I have is that when PBMC's become activated using an activation compound their cellular impedance has been shown to change ~5x in papers [1]. It has also been shown that many rheumatic diseases and infections change the threshold at which PBMC's activate [3]. Karl Morten highlighted that PBMC's left in a tube start to become activated as part of his work on the Accumen mitochondrial function test failed replication [2]. This would seem to indicate that very fresh blood may be required for the plasma swap experiments if they are using immune cells.

Last week I emailed Alain Moreau to see if he has tried the same salt test on PBMC's + plasma using his CellKey cellular impedance analyzer but have not heard back yet. At the Stanford symposium he presented using the CellKey to test drugs by looking for a change in impedance of Jurkat cells soaked in patient plasma. If I understand right, Jurkat cells are a similar size to PBMC's.
MoreauJurkat.JPG

I did also suggest to Alain Moreau about using Lymphoblast cells similar to Paul Fisher, that way the experiments would use patient cells rather than purchased/grown cell lines.

EDIT : Add references
1. Activated PBMC cellular impedace - see supplemental fig 1
https://www.ncbi.nlm.nih.gov/pubmed/23483079
2. Activation of PBMC's over time observation by Karl Morten
https://www.meassociation.org.uk/wp...al-Function-in-MECFS-Acumen-Test-21.08.19.pdf
3. "Bioenergetics of immune cells to assess rheumatic disease activity and efficacy of glucocorticoid treatment"
https://ard.bmj.com/content/62/2/133

EDIT2: In the s4me Karl Morten Q&A videos released this week I did suggest in the questions that he look at cellular respiration using the same salt solution + plasma + cells to see if oxygen consumption differences can be seen on different equipment to the nanoneedle. He said in the video that he has put this on his to do list. This should be a fairly simple experiment. If patient cells + plasma are working harder to remove salt than healthy cells + plasma then that may be detectable by the change in cellular respiration when the salt is added. He did also add that type of cell + collection & storage method of plasma did affect his cellular respiration tests and he wants to get a better handle on what those variables are.
 
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nandixon

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This would seem to indicate that very fresh blood may be required for the plasma swap experiments if they are using immune cells.
Under the scenario I gave, the patient's blood cells would be discarded at the time of the blood draw. The patient's plasma would then be sent to Stanford and tested on freshly prepared cells from a healthy donor. I'm hoping that the plasma itself doesn't need to be fresh, per se, but just needs to be processed and stored in such a way as to keep the "something in the blood" intact.

[For other people: The plasma exchange experiments are implying that the patient's actual cells are not necessary for testing purposes because in the nanoneedle they act just like healthy cells when bathed in healthy plasma. Therefore the researchers should seemingly be able to substitute healthy cells for ME/CFS cells for purposes of testing plasma from an ME/CFS patient in the nanoneedle to see if an abnormal (i.e., increased) impedance signal is given.]

I'll try to locate and post an example of a plasma storage procedure that I saw that I'm hoping can be used to allow patients to send in just the plasma only.
 

nandixon

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This is what I'm hoping might work:

Optimized exosome isolation protocol for cell culture supernatant and human plasma
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4507751/

Twenty millilitres of blood was obtained from a healthy volunteer in EDTA-coated tubes and allowed to sit at room temperature for 30 minutes. Whole blood was then centrifuged at 1,200 g for 10 minutes at 4°C to separate plasma. Plasma was transferred to a clean tube and centrifuged again at 1,800 g for 10 minutes at 4°C before being aliquoted, snap frozen on dry ice and stored at −80°C until use.

(The plasma would be shipped to Stanford surrounded by dry ice.)

And again, we don't know if an exosome is involved or not. I'm just hoping that same procedure would keep whatever the "something in the blood" is viable until tested.
 

nandixon

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EDIT2: In the s4me Karl Morten Q&A videos released this week I did suggest in the questions that he look at cellular respiration using the same salt solution + plasma + cells to see if oxygen consumption differences can be seen on different equipment to the nanoneedle. He said in the video that he has put this on his to do list. This should be a fairly simple experiment. If patient cells + plasma are working harder to remove salt than healthy cells + plasma then that may be detectable by the change in cellular respiration when the salt is added. He did also add that type of cell + collection & storage method of plasma did affect his cellular respiration tests and he wants to get a better handle on what those variables are.
Thanks for suggesting that!
 

FMMM1

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@nandixon We have to also remember the researcher who worked on the nanoneedle has moved to a new position at UC Irvine so perhaps that has slowed down the work, even if he is still involved. It's taken 2 years to test 40 patients so it is clearly a slow process and the nanoneedle needs improved for throughput.

One concern I have is that when PBMC's become activated using an activation compound their cellular impedance has been shown to change ~5x in papers [1]. It has also been shown that many rheumatic diseases and infections change the threshold at which PBMC's activate [3]. Karl Morten highlighted that PBMC's left in a tube start to become activated as part of his work on the Accumen mitochondrial function test failed replication [2]. This would seem to indicate that very fresh blood may be required for the plasma swap experiments if they are using immune cells.

Last week I emailed Alain Moreau to see if he has tried the same salt test on PBMC's + plasma using his CellKey cellular impedance analyzer but have not heard back yet. At the Stanford symposium he presented using the CellKey to test drugs by looking for a change in impedance of Jurkat cells soaked in patient plasma. If I understand right, Jurkat cells are a similar size to PBMC's.
View attachment 34742

I did also suggest to Alain Moreau about using Lymphoblast cells similar to Paul Fisher, that way the experiments would use patient cells rather than purchased/grown cell lines.

EDIT : Add references
1. Activated PBMC cellular impedace - see supplemental fig 1
https://www.ncbi.nlm.nih.gov/pubmed/23483079
2. Activation of PBMC's over time observation by Karl Morten
https://www.meassociation.org.uk/wp...al-Function-in-MECFS-Acumen-Test-21.08.19.pdf
3. "Bioenergetics of immune cells to assess rheumatic disease activity and efficacy of glucocorticoid treatment"
https://ard.bmj.com/content/62/2/133

EDIT2: In the s4me Karl Morten Q&A videos released this week I did suggest in the questions that he look at cellular respiration using the same salt solution + plasma + cells to see if oxygen consumption differences can be seen on different equipment to the nanoneedle. He said in the video that he has put this on his to do list. This should be a fairly simple experiment. If patient cells + plasma are working harder to remove salt than healthy cells + plasma then that may be detectable by the change in cellular respiration when the salt is added. He did also add that type of cell + collection & storage method of plasma did affect his cellular respiration tests and he wants to get a better handle on what those variables are.
Haven't got to watch the videos from the OMF Symposium; hoping to i.e. when they become available. Interesting that activation/maturation increase impedance. If the --- something in the blood - causes maturation then that might give a clue i.e. what the "something in the blood" is. I'm baffled by much of this; hopefully Kark, Ron --- will get funding so that they can clarify what's going on here.
 

nandixon

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Haven't got to watch the videos from the OMF Symposium; hoping to i.e. when they become available.
A video of the Symposium in two parts was made available on Facebook. I don't have a link anymore. Note that there is no mention of the something in the blood so far as I saw. (I skipped over some speakers though.)

Interesting that activation/maturation increase impedance. If the --- something in the blood - causes maturation then that might give a clue i.e. what the "something in the blood" is.
There are many things that can increase cellular impedance. I can't rule it out completely but immunological activation of the cells by the "something in the blood" seems less likely to me because the increase in nanoneedle impedance doesn't happen when cells are simply placed in ME/CFS plasma, but rather only after those cells are additionally stressed with salt. (But maybe the nanoneedle isn't configured to be able to see an initial activation, and only shows the effect of prior cell activation when salt is added.)
 
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Note that there is no mention of the something in the blood so far as I saw.
FYI - Alain Moreau in his talk at the Symposium gave his version of "something in the blood", and how the cellular impedance response of Jurkat cells bathed in patient plasma to various stimuli, measured on his CellKey instrument, may vary by mi-RNA sub group of the patient plasma. Presentation & graphs brought up a lot of questions in my mind as to what he is actually seeing........... As far as I am aware this is the 5th group to identify "something in the blood".

This slide testing with plasma from control & 5 subgroups was one example
MoreauJurkat3.JPG
 
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Moof

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Enjoying following this discussion. Re. the earlier comments about the timing of sample processing, and bearing in mind that my aptitude for maths and science is such that I've never even mastered long division: even if the 'something in the blood' is a lab artefact, it's a different artefact to that produced by healthy controls. It doesn't help us find out what's behind it, but – assuming that the HC samples weren't processed in a much shorter time frame, which would be easy to establish – it still needs explanation. I really feel hopeful that we're closing in on something, despite the enormous technical challenges.
 

FMMM1

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A video of the Symposium in two parts was made available on Facebook. I don't have a link anymore. Note that there is no mention of the something in the blood so far as I saw. (I skipped over some speakers though.)


There are many things that can increase cellular impedance. I can't rule it out completely but immunological activation of the cells by the "something in the blood" seems less likely to me because the increase in nanoneedle impedance doesn't happen when cells are simply placed in ME/CFS plasma, but rather only after those cells are additionally stressed with salt. (But maybe the nanoneedle isn't configured to be able to see an initial activation, and only shows the effect of prior cell activation when salt is added.)

Thank you both @nandixon @ljimbo423 for the heads up about the videos.

I think you're correct about the addition of the salt i.e. the cells with ME plasma have the same impedance signal until the cells are forced to do work (pump salt out of the cell). So I think(!) that suggests the change in impedance isn't simply maturation.
 

FMMM1

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Enjoying following this discussion. Re. the earlier comments about the timing of sample processing, and bearing in mind that my aptitude for maths and science is such that I've never even mastered long division: even if the 'something in the blood' is a lab artefact, it's a different artefact to that produced by healthy controls. It doesn't help us find out what's behind it, but – assuming that the HC samples weren't processed in a much shorter time frame, which would be easy to establish – it still needs explanation. I really feel hopeful that we're closing in on something, despite the enormous technical challenges.
Sounds like a pretty good suggestion i.e. what's the difference in healthy control plasma versus ME plasma. Regarding the challenge in identifying what causes something in the blood, Ron Davis spoke of the body using the same chemical for different purposes --- that makes it more difficult to identify what's causing the change. I think Ron used the example of ATP inside the cell is used for energy production; outside the cells it is used for signalling (purinergic receptors).
 
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@wigglethemouse Thank you for contacting Dr Moreau regarding his work with plasma and salt stress. A website that I find really helpful when I forget the type of cell line being used, such as the Jurkat cells is:
https://www.atcc.org/
It's quite informative, so for Jurkat, you can see that they are from a human who had acute T cell leukemia. So the cells are immortalized and they can be passed over and over, without a lot of apoptosis.