Metabolic profiling reveals anomalous energy metabolism and oxidative stress pathways

[JaimeS]

Hi all,

Thank you for the interest in this article I wrote, I was looking up whether it had been published yet and I noticed this forum came up. Been looking through the forum and it's refreshing to see such a positive collective of people helping one another like this.
I'm more than happy to answer any questions about the work and discuss some of the thoughts in better detail. I will scan back through the posts over time to answer questions that have already been asked.

Cheers,

Chris

Chris,

If you're open to hearing, I did note a mistake. (I was looking at this article a great deal!) I already sent this via 'send researcher a message' on some Research Gate-like site, but I have no idea how / if you'll ever get it.

In the article, you state that there are six metabolites off in the absolute blood metabolites, and six off in the relative. However, in one case, you then go on to list seven rather than six metabolites:

....there were again, six metabolites that were significantly different (Fig. 3a). For this analysis aspartate and glucose were increased, whereas acetate, glutamate, hypoxanthine, lactate, and phenylalanine decreased in ME/CFS patients.

Acetate is the one that is incorrect; Figure 3a shows that acetate is not increased in the relative blood metabolites, just in the absolute.

Thank you so much for this study! I find it very interesting (obviously!)

-J

 

[South]

@JaimeS If you want Chris to see your post, shouldn't you put the "@" symbol in front of his name (to put an "alert" in his phoenix rising In box)?

It would be great if he gets your info!

 

[JaimeS]

@JaimeS If you want Chris to see your post, shouldn't you put the "@" symbol in front of his name (to put an "alert" in his phoenix rising In box)?

It would be great if he gets your info!

I think quoting his post will alert him?

-J

 

[ChrisArmstrong]

One of the issues I think needs explanation, @ChrisArmstrong , is why so many studies point at high lactate in serum, spinal fluid and even in vitro muscle cells, if the problem is poor glycolysis, which I realize is not necessarily an implication of the paper.

Poor glucose utilization, perhaps as a result of a defective insulin response, does not necessarily imply poor glycolysis in terms of mechanism, just output.

There is also a question of what is happening in different tissues, or in tissue versus individual cells.

This study may have involved resting patients as well. Post exercise may lead to different results.

On that theme I find these results consistent, presuming that the samples were taken at rest. At rest we tend to be alkaline, at exercise we are highly acidic. I wonder if this implies different biochemistry at rest and during activity than expected.

A post exercise version of this testing would seem to be the next logical step, or perhaps both pre- and post- exercise (or mental activity) with a comparison of results, versus controls. I realize this requires funding and organization for a disease that is close to blatantly ignored by funding bodies.

There is always a question of circadian variation in these patterns as well, particularly since a large number of us have highly disturbed circadian regulation.

Hi Alex,

Well I haven't actually seen so many recent studies that have indicated an increase of lactate in the blood. In fact, I've seen more of consistent decrease in recent studies: this one in 2010 (Nitric Oxide Metabolite Production During Exercise in Chronic Fatigue Syndrome: A Case-Control Study) and this patent in 2013: Biomarker for fatigue, and use thereof US 20130210045 A1.

I'm aware of the increase found in the CSF, I personally believe the disconnect between the two is extremely important as studies have shown that plasma lactate and CSF lactate usually correlate.
I have read that people with ME/CFS will have an accelerated glycolysis during exercise and it's possible that an accelerated glycolysis could also occur during hyper-activity in the brain. Much easier to control the resting the body than the brain in experiments. Though I wouldn't imagine this is just it, it could well be an issue of transport at the blood brain barrier, I hope they look further into this.

In this paper I discussed the results we had and what conclusion they led to. The data implied a reduced use of glycolysis and a reduced level in some amino acids that highlighted a path by which the TCA cycle could still run (albeit less efficiently). Didn't go too indepth into theories because there is no data to back it up yet and it's not pertinent to do this in an analytical journal. We will come out with a hypothesis paper at some stage but we really need to do further studies. What I will say is that metabolism will change under certain conditions and with purpose. That is, we saw under utilisation of glycolysis at rest (aerobic), but that could be because the body is compensating for the fact that a faster glycolysis acceleration occurs during anaerobic conditions (these fluctuations in metabolism usually relate to alterations in pH, the bigger the fluctuations in pH then the bigger the changes in metabolism under different stressors). Alternatively, the reduced use of glycolysis could be limited to reduce side-effects or limited to promote the production of other metabolites. Molecule/metabolites serve two major purposes in the body, they are the building blocks of matter and they are used to create energy. Generally, if you are fatigued it is because the metabolites are being used to create matter over energy. This is the basic premise from which we work and using Occam's razor I'd suggest that the matter that is being created in ME/CFS is for the creation of immune cells.

Metabolism is extremely complex and dynamic, we think there is value in looking at how it changes under different conditions in people ME/CFS and so you hit the nail on the head when you mentioned exercise studies. That's exactly where I'm at right now, applying for grants so that we can use this method to follow people with ME/CFS over a time-course and monitor the fluctuation in their metabolism when they encounter different stressors (exercise, diet changes, altered states, illness etc). You have to start somewhere in science, this study was done as an information-seeking exercise and a proof of concept for the use of metabolomics in studying ME/CFS while trying to build credibility in the metabolomics field (not many studies have tried too correlate two biofluids). Funding is difficult for ME/CFS alone but by branching out in two fields we can get more general interest into our work and hopefully better opportunities for grants.

I hope this helps answer some questions, though I'm sure it will create new ones. I can try use analogies if people need them to help understand concepts.

-Chris

 

[ChrisArmstrong]

Chris,

If you're open to hearing, I did note a mistake. (I was looking at this article a great deal!) I already sent this via 'send researcher a message' on some Research Gate-like site, but I have no idea how / if you'll ever get it.

In the article, you state that there are six metabolites off in the absolute blood metabolites, and six off in the relative. However, in one case, you then go on to list seven rather than six metabolites:



Acetate is the one that is incorrect; Figure 3a shows that acetate is not increased in the relative blood metabolites, just in the absolute.

Thank you so much for this study! I find it very interesting (obviously!)

-J

Hi Jaime,

Thank you very much for finding and alerting us to that. I didn't receive an email but correspondence probably went to my supervisor. I'll ask him him and see if he corrected it. I can see that you are very interested in the work, do you have a background in science? I myself was blown away by the data that we got and I almost couldn't believe that such a large difference was observed between the ME/CFS and non-ME/CFS groups. There should have been the raw data attached to the article so that others can view it. Did you see it?

-Chris

 

[JaimeS]

I can see that you are very interested in the work, do you have a background in science?

It's late enough in my time-zone that I'm picturing this as the backdrop of a play about science.

My science experience is a mixed bag. BS Rutgers, Plant Science, MS random herbal medicine school you've never heard of, National Boards Certification in Chemistry, many years teaching science - currently Orgo online.

I don't believe I saw the raw data, @ChrisArmstrong, just the tables and such.

-J

 

[JaimeS]

Well I haven't actually seen so many recent studies that have indicated an increase of lactate in the blood.

In exercise tests, our blood lactate tends to be high. My arterial lactate was something like 3x the highest normal value. Whether that's the case at rest remains to be seen; but at rest, my anion gap is high. n=1, but still.

that could be because the body is compensating for the fact that a faster glycolysis acceleration occurs during anaerobic conditions (these fluctuations in metabolism usually relate to alterations in pH, the bigger the fluctuations in pH then the bigger the changes in metabolism under different stressors).

I've never heard it put quite that way, @ChrisArmstrong. It should be mentioned that - again, anecdotally - a lot of us seem to have reactive hypoglycemia, with blood sugar too high sometimes, and too low others.

Generally, if you are fatigued it is because the metabolites are being used to create matter over energy.

As a chemist, this statement makes my brain hurt.

by branching out in two fields we can get more general interest into our work and hopefully better opportunities for grants.

Excellent point!

-Jaime

 

[ChrisArmstrong]

It's late enough in my time-zone that I'm picturing this as the backdrop of a play about science.

My science experience is a mixed bag. BS Rutgers, Plant Science, MS random herbal medicine school you've never heard of, National Boards Certification in Chemistry, many years teaching science - currently Orgo online.

I don't believe I saw the raw data, @ChrisArmstrong, just the tables and such.

-J

Haha, who wouldn't love a musical about science. I'd go just to see them dance out a PCR reaction.

Yeah the raw data is meant to be attached to the journal online. It's part of the metabolomics journal to have a data deposition at the bottom of the page. It's a link to a site called metabolights.

 

[JaimeS]

It's a link to a site called metabolights.

Oooooh.

Also, a musical about science would be AWESOME. <3

-J

 

[ChrisArmstrong]

In exercise tests, our blood lactate tends to be high. My arterial lactate was something like 3x the highest normal value. Whether that's the case at rest remains to be seen; but at rest, my anion gap is high. n=1, but still.



I've never heard it put quite that way, @ChrisArmstrong. It should be mentioned that - again, anecdotally - a lot of us seem to have reactive hypoglycemia, with blood sugar too high sometimes, and too low others.



As a chemist, this statement makes my brain hurt.



Excellent point!

-Jaime

I don't know how to just tag a name so I'm replying.

Yeah, the exercise tests seemed to indicate an accelerating glycolysis that produces lactate. Anecdotally people will experience different things, I'm not expecting to grab a single cause for such a complicated illness but understand a broad mechanism that may be in play. Metabolite levels are different from person to person and are impacted by a myriad of factors. The pH fluctuation in people with ME/CFS interest me greatly, I think bicarbonate is of particular importance in ME/CFS because if the reduced level of ATP from mitochondria does occur then it would lower the level of CO2 and thus bicarbonate.

Haha yeah i'm definitely not using science language there. Energy is not created by metabolites, they are a source of energy that is released when their bonds are broken. By matter I mean they form biological mass.
The best example of this is the Warburg effect if you want to read more. Glycolysis is used for quick energy over efficiency and along with glutaminolysis you create a bunch of metabolites that are required to build a cell. This process purposefully stops the use of the TCA cycle because although it is far more energy efficient it is also creates CO2, which essentially removes carbon from the cells (an essential part of biological mass).

 

[JaimeS]

To tag someone, you just use the at symbol followed by the name below their picture, thus: @ChrisArmstrong . Usually, the computer will autocomplete it after you've typed a few letters.

So would the implication be accelerated cell turnover due to oxidative stress? Which causes TCA cycle to slow, to save carbon?

Hmmmm. That's certainly an interesting thought... Bed now, though!

-J

 

[ChrisArmstrong]

To tag someone, you just use the at symbol followed by the name below their picture, thus: @ChrisArmstrong . Usually, the computer will autocomplete it after you've typed a few letters.

So would the implication be accelerated cell turnover due to oxidative stress? Which causes TCA cycle to slow, to save carbon?

Hmmmm. That's certainly an interesting thought... Bed now, though!

-J

@JaimeS I would think accelerated cell turnover would be due to oxidative stress but also other factors. The proliferation of immune cells for a chronically activated immune system would be a factor, pH would be a factor, periods of fever and the expulsion of cellular mass that follows. Many reasons why people with ME/CFS may have an accelerated cell turnover. What isn't clear is whether these results are suggesting cell turnover because this data showed that glycolysis appeared to be inhibited. When I say appeared, I say it because it could be the complete opposite. It may be that the body is so well adapted to dealing with accelerated glycolysis that it may have reset it's baseline levels at rest to accommodate a larger than average change during anaerobic periods (this would actually backup the current mitochondrial dysfunction hypothesis - i didn't include it because it's too much of a leap to make right now). That's why it's important that we look at the change over time in longitudinal studies.

 

[alex3619]

It may be that the body is so well adapted to dealing with accelerated glycolysis that it may have reset it's baseline levels at rest to accommodate a larger than average change during anaerobic periods (this would actually backup the current mitochondrial dysfunction hypothesis - i didn't include it because it's too much of a leap to make right now).

This is my suspicion based on our resting lactate versus exercising lactate. We are too alkaline at rest, and too acidic in activity, based on Julia Newtons data.

 

[ChrisArmstrong]

This is my suspicion based on our resting lactate versus exercising lactate. We are too alkaline at rest, and too acidic in activity, based on Julia Newtons data.

Which data are you talking about specifically, can you link me?

 

[alex3619]

Which data are you talking about specifically, can you link me?

This was, I think, from a video presentation of her data last year, but I could be wrong. It was at a UK conference or something. I am not sure this has been formally published. The best way might be to contact her in the UK. At least one of her muscle test bed studies, on electrically stimulated muscle cells in vitro, has been published.

 

[ChrisArmstrong]

This was, I think, from a video presentation of her data last year, but I could be wrong. It was at a UK conference or something. I am not sure this has been formally published. The best way might be to contact her in the UK. At least one of her muscle test bed studies, on electrically stimulated muscle cells in vitro, has been published.

Thank you, I will look it up. I also think that a move from ATP creation to cell proliferation is occurring here but not ready to publish this theory in scientific journals. This work in this paper of ours is trying to provide a framework to link all this together. It needs to be clear that it is cell proliferation and from there we can try to ascertain the cause because it still may be a range of any other possibilities. I've also done some work on the gut with this same cohort, I'm looking through this work now and hopefully it will shed some light on its involvement. After this I might release a hypothesis paper.

 

[SOC]

Fascinating way to put it, @ahmo . Is there a cat inside?

-J

And is it dead or alive ?

Did you ever wonder what would happen if there were two cats in Schrodinger's box (to say nothing of @ahmo's body)?

 

[halcyon]

@ChrisArmstrong I was curious why this study was only done on women. Sorry if this is answered in the full paper I haven't been able to read it.

 

[ChrisArmstrong]

@ChrisArmstrong I was curious why this study was only done on women. Sorry if this is answered in the full paper I haven't been able to read it.

One gender was chosen to minimise gender-based differences that may confound metabolites differences. Chose women because there are more of them with ME-CFS.

 

[alex3619]

And is it dead or alive ?

Did you ever wonder what would happen if there were two cats in Schrodinger's box (to say nothing of @ahmo's body)?

Two zombies instead of one? Both dead AND alive? Maybe they have ME.