Metabolic profiling indicates impaired pyruvate dehydrogenase function in myalgic encephalopathy/CFS (Fluge et al., 2016)

[lansbergen]

But I think it's too early to discount the persistent infection hypothesis entirely. Agree to disagree, of course!

I agree

 

[Hip]

Some important info for anyone interested in ME/CFS energy metabolism dysfunction:

A likely explanation has been found for the apparent contradiction in results from the Lawson et al study (which found higher than normal ATP levels the cells of ME/CFS patients), and the Myhill, Booth and McLaren-Howard study (which found lower than normal ATP levels the cells of ME/CFS patients).

In essence, the explanation is this: since the mitochondrial blocking factor that negatively affects the mitochondria in ME/CFS appears to be found in the blood serum (Fluge and Mella's results demonstrated this), unless you test cells that are directly extracted from ME/CFS patients, you won't see the effects of this blocking factor.

In the case of the Lawson study, they did not use cells directly extracted from ME/CFS patients, but rather cells that were taken from ME/CFS patients and then cultured in vitro, and were thus not really exposed to the blocking factor, so these cultured cells then appeared healthy. This is likely the basis of the contradictory results, and the explanation of why Lawson et al did not find a reduced energy metabolism in ME/CFS.

More info in this post.


This is an important point to bear in mind for any future research testing the energy metabolism of ME/CFS patients cells: if you are not using cells taken directly from ME/CFS patients, but instead grow them in culture, you will likely not be able to observe the energy metabolism dysfunction.

 

[deleder2k]

I tagged @Ben Howell in a previous post a couple pages ago here, and this is @deleder2k's thread, who I believe is actually in the current Phase 3 rituximab trial in Norway, so hopefully he'll ask Fluge/Mella about testing for S1P levels and looking for autoantibodies to the S1P receptors and to S1P itself.

I won't be going to Haukeland for sometime I'm afraid. I am sure both Fluge and Mella would love to hear more about this. Do send them an e-mail about this!

 

[nandixon]

.

Some important info for anyone interested in ME/CFS energy metabolism dysfunction:

A likely explanation has been found for the apparent contradiction in results from the Lawson et al study (which found higher than normal ATP levels the cells of ME/CFS patients), and the Myhill, Booth and McLaren-Howard study (which found lower than normal ATP levels the cells of ME/CFS patients).

Here's a slightly modified explanation for this under the sphingosine-1-phosphate (S1P) hypothesis. (The S1P hypothesis is that in ME/CFS there is impaired S1P signaling due to either abnormally low levels of S1P or because there are autoantibodies against S1P receptors.)

The major source of S1P in the blood is erythrocytes (RBCs), which act as a sort of storage vehicle for S1P. When released from the RBCs, S1P is bound to albumin and other plasma materials for transport to the S1P receptors on cell surfaces. (Reference; see also this reference under the "Sphingosine 1-Phosphate in the Blood" section.)

In the Lawson study, the cells being cultured were PBMCs (peripheral blood mononuclear cells), which do not contain or produce any significant amount of S1P - because there are no RBCs present.

Thus, the Lawson study cells never actually had any significant amount of the "blocking factor" (e.g., low S1P in the case of the patients) to begin with, and what was there at the start would have been diluted after culturing.

This put the patient cells and the control cells on essentially an equal footing after being cultured. Except the patient cells presumably would have had various upregulations built into their gene expressions to compensate for the impaired S1P signaling they had previously been subjected to, thus explaining the higher ATP that Lawson found in the patient cells.

On the other hand, Myhill used neutrophils directly without culturing. Those cells' receptors would have been primed with S1P immediately following their isolation, and the patient cells would have been primed with less S1P than the controls. (Note that RBCs are typically lyased during isolation of neutrophils, releasing additional S1P.) Or alternatively, there were autoantibodies still present in the case of the patient cells.

So in the Myhill study, the patient cells were subjected to impaired S1P signaling. This causes mTORC1 to not be properly activated (among other problems) and leads to inhibition of the PDH complex as found by Fluge and Mella, and the lower ATP found by Myhill.

 

[alicec]

I tend to think that all the symptoms of this illness are fundamentally caused by the same problem: lack of energy.

Yes I'm inclined to agree with you, I was just trying to think of a reason for the lack of effect of allithiamine on brain fog.

You say that sulbutiamine did help with this. Maybe it is just dose related since sulbutiamine appears to be more potent, ie its molecular structure means that it achieves even higher intracellular levels of active B1 than allithiamine, albeit by the same mechanism.

 

[Sasha]

This is an important point to bear in mind for any future research testing the energy metabolism of ME/CFS patients cells: if you are not using cells taken directly from ME/CFS patients, but instead grow them in culture, you will likely not be able to observe the energy metabolism dysfunction.

But didn't Julia Newton's team find problems in cultured muscle cells from ME patients that were "excercised" by an electrical stimulus (although I seem to remember that this didn't replicate, though I'm not sure)?

 

[Hip]

But didn't Julia Newton's team find problems in cultured muscle cells from ME patients that were "excercised" by an electrical stimulus (although I seem to remember that this didn't replicate, though I'm not sure)?

Good point; I think this may be the Julia Newton study you are referring to.

Though I can see a likely explanation, based on what McLaren-Howard said: it's to do with the number of generations of new cells (cell replications) that you grow in your cell culture.

In Dr John McLaren-Howard's original note on this subject, he says that the more new generations of cells you grow in your cell culture (ie, the more cell replications you have in your culture dish), the more it dilutes down the concentration of the mitochondrial blocking factor in these cells (the mitochondrial blocking factor that is assumed to be in the blood of ME/CFS patients).

This is because when you first extract cells from an ME/CFS patient (cells from the blood or tissues), obviously these cells have been bathing within the ME/CFS patient's blood within his body, and so these cells will contain the full concentration of blocking factor, and will thus have a dysfunctional energy metabolism.

But now when you begin growing your culture of cells, starting with seed cells that you extracted from the patient, each new replication of the cells (which occurs by cell division, creating two daughter cells from the original cell) will obviously half the concentration of the blocking factor inside the cell, because the same amount of blocking factor that you had in one cell now has to be shared between its two daughter cells.

So each time a cell replicates in your cell culture dish, the concentration of the blocking factor halves. Thus if for example an original cell has replicated 5 times, your blocking factor concentration inside the cells goes from 100%, down to 50%, then down to 25%, then 12.5%, then 6%, and finally 3%.

So after just 5 new generations of cells, the original blocking factor concentration in the cell goes from 100% down to just 3%, and so the cells then start functioning normally, as most of the blocking factor has gone.

So the crucial issue in any cell culture experiment is the number of new generations of cells that you grow in the culture dish. The more generations you grow, the more cell replications, the more the ME/CFS blocking factor gets diluted down and washed out, and the more the cells will return to normal healthy functioning.



I am not sure of the exact experimental procedure used in the Lawson et al study, but if in their cell culture they grew many generations of cells, this would have washed out nearly all of the blocking factor.

Likewise, I don't know the experimental setup used in Julia Newton et al study, but if they grew very few new cells in culture (perhaps because they already had extracted sufficient cells from the patients' muscles), then the blocking factor would not have been washed out, and would have remained in high concentration within the cells in the culture, so in which case, the cells would have maintained their original ME/CFS dysfunctional energy metabolism.

 

[Sasha]

@Hip, I haven't been reading this thread with the care it deserves so this might have come up, but is it likely that the researchers are aware of this issue and the possible effects on replicability? Should we be writing and telling them?

 

[Hip]

@Hip, I haven't been reading this thread with the care it deserves so this might have come up, but is it likely that the researchers are aware of this issue and the possible effects on replicability? Should we be writing and telling them?

It's quite possible that other researchers may not be aware of this, as the notion that the ME/CFS dysfunctional energy metabolism may be caused by a blocking factor in the blood is, as far as I am aware, a not a widely understood or recognized idea / discovery.

This idea of blocking factor in the blood is implicit in the 2009 and 2012 ME/CFS energy metabolism studies of Myhill, Booth and McLaren-Howard, and the earliest reference to such a concept that I know of is found in a 1985 study by Professor Peter Behan, who suggested that an anti-mitochondrial antibody in the blood (arising from viral infection) could be the cause of the energy metabolism dysfunction of ME/CFS.

And of course now with the very incisive 2016 study by Fluge and Mella discussed in this thread, who exposed healthy myoblast cells from healthy people to the blood serum of ME/CFS patients, and found that this led to alterations in the energy metabolism of those cells, we have clearcut evidence that there is some factor in the serum of ME/CFS patients that affects cellular energy metabolism.


I certainly think it would worth writing to and pointing all this out to any researcher who we hear is going to conduct a new study on the cells of ME/CFS patients.

 

[Sasha]

It's quite possible that other researchers may not be aware of this, as the notion that the ME/CFS dysfunctional energy metabolism may be caused by a blocking factor in the blood is, as far as I am aware, a not a widely understood or recognized idea / discovery.

This idea of blocking factor in the blood is implicit in the 2009 and 2012 ME/CFS energy metabolism studies of Myhill, Booth and McLaren-Howard, and the earliest reference to such a concept that I know of is found in a 1985 study by Professor Peter Behan, who suggested that an anti-mitochondrial antibody in the blood (arising from viral infection) could be the cause of the energy metabolism dysfunction of ME/CFS.

And of course now with the very incisive 2016 study by Fluge and Mella discussed in this thread, who exposed healthy myoblast cells from healthy people to the blood serum of ME/CFS patients, and found that this led to alterations in the energy metabolism of those cells, we have clearcut evidence that there is some factor in the serum of ME/CFS patients that affects cellular energy metabolism.


I certainly think it would worth writing to and pointing all this out to any researcher who we hear is going to conduct a new study on the cells of ME/CFS patients.

@Jonathan Edwards - do you think that researchers are aware that a possible reason for some studies not replicating might be this issue (which I don't trust myself to summarise)? I wonder if it's worth a bit of a re-review of past work.

 

[nandixon]

I won't be going to Haukeland for sometime I'm afraid. I am sure both Fluge and Mella would love to hear more about this. Do send them an e-mail about this!

I just sent an email to Dr Fluge. I wrote the following:

Dear Dr. Fluge,

I'm a ME/CFS patient of nearly 20 years and I wanted to first express my deep gratitude for the work that you and Dr. Mella are doing and for the care and concern you show towards your patients.

It seems apparent to me, as a former researcher in medicinal chemistry (and having undergone many tests myself to try to understand this disease), that your most recent study showing impaired PDH complex functioning is indicating that the mTORC1 pathway is not being properly activated in ME/CFS.

I write on the Phoenix Rising ME/CFS forum under the name "nandixon" and I first posted about this here: http://forums.phoenixrising.me/inde...-encephalopathy-cfs.48446/page-12#post-800136

I then found that your study fits very well with that of Dr. Naviaux and his finding of low ceramides to suggest that there is impaired sphingosine-1-phosphate (S1P) signaling in ME/CFS to cause the inactivity of mTORC1. I wrote about this here: http://forums.phoenixrising.me/inde...-encephalopathy-cfs.48446/page-15#post-801205

Have you considered measuring the levels of S1P in ME/CFS? It doesn't appear that any researchers have ever done this.

If S1P is low, then presumably it's because of the low ceramides (or because there are autoantibodies directed against S1P). In this case, rituximab and cyclophosphamide may be working by resetting the sphingomyelin cycle (perhaps by breaking a negative feedback loop that includes disregulated signaling by B- and/or T-cells) to improve, for example, the activity of acid sphingomyelinase and increase ceramides.

If S1P is high (or normal), then perhaps it's because there are autoantibodies directed against one or more S1P receptors (e.g., S1PR1 aka S1P1), causing S1P production from ceramides to be upregulated in a effort to overcome this (and making ceramides low in the process).

Thank you for considering this, and many thanks again for your work.

 

[eljefe19]

@nandixon Thanks! Very well written.

 

[trails]

@nandixon Thank you for passing along your insights to Dr. Fluge!

 

[Snow Leopard]

I won't be going to Haukeland for sometime I'm afraid. I am sure both Fluge and Mella would love to hear more about this. Do send them an e-mail about this!

Are they open to discussing with (educated) patients? (via unsolicited email?)

Also, has anyone been able to talk with @znahle via email? I don't seem to be getting any replies. It would be interesting to hear his thoughts on this study and the various hypotheses.

 

[Snow Leopard]

@nandixon If they need more convincing.

Endothelial functions of sphingosine-1-phosphate.
https://www.ncbi.nlm.nih.gov/pubmed/20502008

The role of sphingosine-1-phosphate in endothelial barrier function
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4169319/

S1P Control of Endothelial Integrity
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4240614/

Sphingosine-1-Phosphate Signaling in Endothelial Disorders.
https://www.ncbi.nlm.nih.gov/pubmed/27115142

etc.

Also:
The role of sphingolipids in the control of skeletal muscle function: a review.
https://www.ncbi.nlm.nih.gov/pubmed/10937863

Additional Edit, also:
Viral infections and sphingolipids.
https://www.ncbi.nlm.nih.gov/pubmed/25525752

Sphingolipids in viral infection.
https://www.ncbi.nlm.nih.gov/pubmed/25525752

Sphingosine-1-Phosphate Rapidly Increases Cortisol Biosynthesis and the Expression of Genes Involved in Cholesterol Uptake and Transport in H295R Adrenocortical Cells
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3508734/

It seems to me that they may have (should have) come across hypotheses along these lines before,when investigating their Endothelial dysfunction hypothesis!

I still think it's a receptor issue, but S1P levels need to be investigated anyway!

There are alternative hypotheses to low ceramides vs S1P, (eg a shift in equilibrium towards S1P) which is why these things need to be investigated directly.

I also wonder about the relationship between anti-ganglioside autoantibodies and sphingolipids...

 

[Sidereal]

Great email @nandixon. Worth letting Naviaux know about this too IMHO.

 

[A.B.]

Question: could decreased ceramides explain the blood sugar issues in a subgroup of patients? Would excessive sensitivity to insulin lead to blood sugar levels that are somewhat unstable and tend to fluctuate too rapidly?

Hormonal
Increased ceramide synthesis leads to both leptin resistance and insulin resistance by increasing SOCS-3 expression.[13] Elevated level of ceramide results in the inhibition of insulin signal transduction pathway and the serine phosphorylation of JNK, leading to insulin resistance.[14]

https://en.wikipedia.org/wiki/Ceramide

 

[Bdeep86]

I've ordered 25 grams of dichloroacetate (DCA) for £32, and will be trying it soon.

I am going to take benfotiamine, alpha lipoic acid and acetyl-L-carnitine with the DCA in order to try to prevent any neuropathy (as recommended by the article quoted in this post).

Other side effects of DCA can include heartburn, nausea, vomiting, indigestion, but these can be countered with a proton pump inhibitor drug (see the same post). I wonder if DCA can be administered as a suppository or transdermally to try to avoid these stomach issues.

The one forum member who tried DCA said:

I want to try to avoid these stomach side effects.

I read that caffeine can boost the effects of DCA, which then allows for lower doses of DCA, and thus lower side effects. However, I have also read articles advising caution when taking caffeine with DCA, perhaps because it boosts the effects of DCA too much. One article said taking caffeine with DCA is more likely to cause the fatigue and weakness side effects.

Daily DCA dose recommendations for cancer treatment that I saw here are 10 to 20 mg per kg body weight. In a study of treatment of congenital lactic acidosis in children, a DCA dose of 12.5 mg per kg body weight was given every 12 hours.

So typical DCA doses would equate to around 1 to 2 grams daily. I may try 300 mg of DCA three times daily to start with.

Can you repost the initial link the article, it didn't work for me when i clicked it. Thanks Hip

 

[junkcrap50]

Can you repost the initial link the article, it didn't work for me when i clicked it. Thanks Hip

Ah yes @Hip , your first link doesn't work.

 

[Cheshire]

Miriam Tucker's coverage of the study (interesting interview, they explain, that according to them, these abnormalities cannot be explained by deconditionning):

http://www.medscape.com/viewarticle/874434#vp_2