Dysregulated provisions of oxidisable substrates to the mitochondria in ME/CFS lymphoblasts (Missailidis et al., 2021)

Pyrrhus

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Dysregulated Provision of Oxidisable Substrates to the Mitochondria in ME/CFS Lymphoblasts (Missailidis et al., 2021)
https://www.mdpi.com/1422-0067/22/4/2046
From Paul Fisher's group in Australia.

Written abstract:
Missailidis et al 2021 said:
Although understanding of the biomedical basis of Myalgic Encephalomyelitis/Chronic Fatigue Syndrome (ME/CFS) is growing, the underlying pathological mechanisms remain uncertain. We recently reported a reduction in the proportion of basal oxygen consumption due to ATP synthesis by Complex V in ME/CFS patient-derived lymphoblast cell lines, suggesting mitochondrial respiratory inefficiency.

This was accompanied by elevated respiratory capacity, elevated mammalian target of rapamycin complex 1 (mTORC1) signaling activity and elevated expression of enzymes involved in the TCA cycle, fatty acid β-oxidation and mitochondrial transport. These and other observations led us to hypothesise the dysregulation of pathways providing the mitochondria with oxidisable substrates.

In our current study, we aimed to revisit this hypothesis by applying a combination of whole-cell transcriptomics, proteomics and energy stress signaling activity measures using subsets of up to 34 ME/CFS and 31 healthy control lymphoblast cell lines from our growing library.

While levels of glycolytic enzymes were unchanged in accordance with our previous observations of unaltered glycolytic rates, the whole-cell proteomes of ME/CFS lymphoblasts contained elevated levels of enzymes involved in the TCA cycle (p = 1.03 × 10−4), the pentose phosphate pathway (p = 0.034, G6PD p = 5.5 × 10−4), mitochondrial fatty acid β-oxidation (p = 9.2 × 10−3), and degradation of amino acids including glutamine/glutamate (GLS p = 0.034, GLUD1 p = 0.048, GOT2 p = 0.026), branched-chain amino acids (BCKDHA p = 0.028, BCKDHB p = 0.031) and essential amino acids (FAH p = 0.036, GCDH p = 0.006).

The activity of the major cellular energy stress sensor, AMPK, was elevated but the increase did not reach statistical significance. The results suggest that ME/CFS metabolism is dysregulated such that alternatives to glycolysis are more heavily utilised than in controls to provide the mitochondria with oxidisable substrates.
(spacing added for readability)


Graphical abstract:
953EE321-7CA4-405B-99CC-8F85A69DC0D3.jpeg
 
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Learner1

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I've had both lab and metabolic testing in a treadmill showing that I'm almost exclusively using glycolysis, such that I burn up all the glycogen stores in my muscles prematurely contributing to exercise intolerance, and that I have trouble with fatty acid oxidation.

As with everything else in this disease, testing and treatment must be individualized to the patient - pronouncements of a general nature contribute to the failure of many treatments when they are inappropriately applied to the wrong problem, unfortunately, contributing to frustration and despair.

However, it is always good to see the new research and compare to one's own situation to glean any insights that might inform one's own treatment.
 

gbells

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We recently reported a reduction in the proportion of basal oxygen consumption due to ATP synthesis by Complex V in ME/CFS patient-derived lymphoblast cell lines, suggesting mitochondrial respiratory inefficiency.
https://www.mdpi.com/1422-0067/22/4...DDjhl05RuAkuX8I_arkUWEAtVMDUapyWfeS8yc9HqCE8g

I told you guys that aerobic respiration in the mitochrondria was inhibited by mitochondral fragmentation from the viruses. Prusty proved that. It is a protective response to limit viral spread. That's why D-ribose improves energy by boosting anaerobic ATP generation.
 

Learner1

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That's why D-ribose improves energy by boosting anaerobic ATP generation.
NAD+ works closer to actual energy production than d-ribose, which has to go through several conversions, with co-factors, to get there.

And:

"Keep in mind that the safety profile of D-ribose is relatively unknown given the lack of well-designed clinical studies. The list of side effects below is not a definite one, and you should consult your doctor about other potential side effects based on your health condition and possible drug or supplement interactions. Seek medical attention if you notice any severe or mild, persistent adverse effects after supplementing with D-ribose.

By inducing protein aggregation and rapidly producing AGEs (advanced glycation end products), D-ribose may be involved in cell dysfunction and cognitive impairments [31, 32].

Long-term oral administration of D-Ribose induces memory loss with anxiety-like behavior and also elevates Aβ-like deposition and Tau hyperphosphorylation associated with Alzheimer’s [33]."

From:
https://selfhacked.com/blog/d-ribose-health-benefits/
 

gbells

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NAD+ works closer to actual energy production than d-ribose, which has to go through several conversions, with co-factors, to get there.

And:

"Keep in mind that the safety profile of D-ribose is relatively unknown given the lack of well-designed clinical studies. The list of side effects below is not a definite one, and you should consult your doctor about other potential side effects based on your health condition and possible drug or supplement interactions. Seek medical attention if you notice any severe or mild, persistent adverse effects after supplementing with D-ribose.

By inducing protein aggregation and rapidly producing AGEs (advanced glycation end products), D-ribose may be involved in cell dysfunction and cognitive impairments [31, 32].

Long-term oral administration of D-Ribose induces memory loss with anxiety-like behavior and also elevates Aβ-like deposition and Tau hyperphosphorylation associated with Alzheimer’s [33]."

From:
https://selfhacked.com/blog/d-ribose-health-benefits/
D-ribose and NAD work through different mechanisms and aren't equivalent.

As the adage goes "the dose makes the poison". I dose d-ribose per appetite along with the immunotherapy regimen so that probably limits the AGEs and other side effects. It probably also is limited by administration of reduced glutathione. I haven't experienced any of the side effects you mentioned thus far through intermittent appetite-based dosing of D-ribose.
 
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Pyrrhus

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Let's remember that there are many other threads to discuss Ribose and NADH in general, but that this is not necessarily the thread for that...

Here is the previous paper from Paul Fisher's group:
An Isolated Complex V Inefficiency and Dysregulated Mitochondrial Function in Immortalized Lymphocytes from ME/CFS patients 9/2019
https://forums.phoenixrising.me/thr...ymphocytes-from-me-cfs-patients-9-2019.77577/

Which originally concluded:
Our results suggest a model in which ME/CFS lymphoblasts have a Complex V defect accompanied by compensatory upregulation of their respiratory capacity that includes the mitochondrial respiratory complexes, membrane transporters and enzymes involved in fatty acid β-oxidation.

However, pursuant to a January 2020 disclosure by the authors, the conclusion has changed from a "defective Complex V" to a "leaky mitochondrial membrane":
https://forums.phoenixrising.me/thr...cfs-patients-9-2019.77577/page-3#post-2323821
 
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Learner1

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Let's remember that there are many other threads to discuss Ribose and NADH in general, but that this is not necessarily the thread for that...

Here is the previous paper from Paul Fisher's group:
An Isolated Complex V Inefficiency and Dysregulated Mitochondrial Function in Immortalized Lymphocytes from ME/CFS patients 9/2019
https://forums.phoenixrising.me/thr...ymphocytes-from-me-cfs-patients-9-2019.77577/

Which concluded:
Do you know of any kind of test that one can verify what one's complex V is doing? And one wonders what complex V in cells that aren't lymphocytes are doing.

I'm still trying to figure out why treadmill testing showed my fatty acid β-oxidation was downregulated, while my labs have shown consistently high glutaric acid and low myoglobin, which occur in impaired fatty acid β-oxidation. My muscles seem to prefer glycolysis.

Has anyone else done metabolic testing on a treadmill?
 

Pyrrhus

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Main points of this paper:
  • 34 ME/CFS and 31 healthy control lymphoblast cell lines were used.
  • The energy-generating TriCarboxylic Acid (TCA) cycle was up-regulated, indicating a high energy demand.
  • However, the rate of Glycolysis, which usually provides substrates for the TCA cycle, was unchanged.
  • The rate of Beta Oxidation, which also provides substrates for the TCA cycle, was increased.
  • The rate of Glutaminolysis, which also provides substrates for the TCA cycle, was increased.
  • The rate of BCAA Oxidation, which also provides substrates for the TCA cycle, was increased.
  • Enzymes which break down phenylalanine, tyrosine, lysine, and tryptophan, perhaps to provide substrates for the TCA cycle, were also increased. (not shown below)
  • The pentose phosphate pathway, which provides substrates for nucleic acid synthesis and for some amino acid synthesis, was also increased. (not shown below)

Here is a diagram that might help:
671CC7E1-EDE2-4173-B943-22B8EEBDF022.jpeg
 
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Learner1

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Lymphoblasts are precursors to B cells and T cells, both of which can be low in ME0CFS patients.

So, again, would this be true for mitos in other cells, like muscle cells or brain cells? I'm not convinced that these findings of n lymphoblasts carry through to the entire body.

I have found isoleucine and leucine, which are BCAAs, to be depleted and taking them to reverse or avoid PEM.

But, my doctors and I think that my mitos preference for glycolysis instead of beta oxidation causes my muscles' glycogen stores to be prematurely depleted, causing exercise intolerance.

I have not found glutamine to be depleted.
 

bread.

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Lymphoblasts are precursors to B cells and T cells, both of which can be low in ME0CFS patients.

So, again, would this be true for mitos in other cells, like muscle cells or brain cells? I'm not convinced that these findings of n lymphoblasts carry through to the entire body.

I have found isoleucine and leucine, which are BCAAs, to be depleted and taking them to reverse or avoid PEM.

But, my doctors and I think that my mitos preference for glycolysis instead of beta oxidation causes my muscles' glycogen stores to be prematurely depleted, causing exercise intolerance.

I have not found glutamine to be depleted.
Interesting though how you think the study has no significant data to show because it is derived from alterations in one specific department (lymphoblasts) and not others (any kind of tissue) while at the same time you find your BCAAs depleted in your blood but not in your tissues.

Inconsistent reasoning with all due respect.

Both your BCAA results and the lymphoblast results show a very small part of what is really going on of course.
 

Pyrrhus

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I have not found glutamine to be depleted.
Glutamine is the most abundant amino acid in the body, so small drops may not be noticed.

In addition, glutaminolysis occurs primarily in rapidly-dividing cells, such as in the intestines or lymphocytes, but not necessarily in all cells. (Glutaminolysis generally occurs when pyruvate is lacking.)

Nonetheless, a few studies found low glutamine in ME/CFS:

The Role of Glutamine in the Aetiology of the Chronic Fatigue Syndrome (Rowbottom et al., 1998)
https://forums.phoenixrising.me/thr...-glutamine-in-the-aetiology-of-the-cfs.10431/
(link only accessible to Phoenix Rising members with at least 100 posts)

NMR metabolic profiling of serum identifies amino acid disturbances in chronic fatigue syndrome (Armstrong et al., 2012)
https://forums.phoenixrising.me/thr...-fatigue-syndrome-armstrong-et-al-2012.20614/
 
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Learner1

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It's not a matter of noticing. Low glutamine just doesn't apply to everyone with ME/CFS.
Screenshot_20210221-205456.png

I've found huge oxidative stress and glutathione depletion to be a problem. Glutamine, cysteine, and glycine are needed to produce glutathione. If one is depleted in glycine and/or cysteine, glutamine won't get used up.
 

Learner1

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Interesting though how you think the study has no significant data to show because it is derived from alterations in one specific department (lymphoblasts) and not others (any kind of tissue) while at the same time you find your BCAAs depleted in your blood but not in your tissues.

Inconsistent reasoning with all due respect.

Both your BCAA results and the lymphoblast results show a very small part of what is really going on of course.
I did not say the study has no significant data to show.

What I'm confused by is that both my labs AND a 55 minute metabolic test on a treadmill showed very clearly that I have extremely impaired fatty acid oxidation and preferentialyly do glycolysis, which seems to be the opposite of what this study says.

Therefore, I wondered if my muscles are doing something different than my lymphoblasts for some reason.

Unfortunately, I realize that very few other patients have had these tests to corroborate or disagree with the study results or provide any kind of further insight. I sure wish there were.

Impaired mitochondria is at the root of many serious diseases, including neurodegenerative diseases and cancer, both of which my parents succumbed to and I have already had cancer. I'm afraid I have a bias toward figuring out how to normalize dysfunctional mitochondria to improve my function as well as to avoid these other awful diseases.

As for BCAAs, I know they were depleted in both blood and urine. No idea about tissues - do you know how to test tissues for aminos? I did find, when I was deficient in isoleucine and leucine, that taking BCAAs after I had PEM could reverse PEM, and taking them before exercise could avoid PEM.
Screenshot_20210221-212445.png

After using this strategy, along with working on increasing protein intake over a year, I found PEM to be rare and the BCAAs less effective when it happened (though glutathione, which I'm still short of is effective). My labs had changed, to the point where BCAA supplementation was no longer needed.
Screenshot_20210221-212519.png

Of course, there's a lot of other stuff going on, both with me and in the studies.
 

Pyrrhus

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And, for good measure, here is a study that found lower BCAA's in ME/CFS compared to controls after an exercise challenge:

Chronic fatigue syndrome: new evidence for a central fatigue disorder. (Georgiades et al., 2003)
https://forums.phoenixrising.me/thr...gue-syndrome-new-evidence-for-a-central.9229/
(only accessible to Phoenix Rising members with at least 100 posts)

Note that BCAA's are typically burned for energy by muscle tissue during exercise, but this study found that they were burned more in ME/CFS patients than they were in controls.
 
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Learner1

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And for good measure, here is a study that found lower BCAA's in ME/CFS compared to controls after an exercise challenge:

Chronic fatigue syndrome: new evidence for a central fatigue disorder. (Georgiades et al., 2003)
https://forums.phoenixrising.me/thr...gue-syndrome-new-evidence-for-a-central.9229/
(only accessible to Phoenix Rising members with at least 100 posts)

Note that BCAA's are typically depleted by muscle tissue during exercise, but this study found that they were depleted in ME/CFS patients more than they were in controls.
That matches my experience. However, this study was on only about a dozen patients selected by what criteria?

Fluge and Mella also found disturbed amino acid metabolism.
 

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Pyrrhus

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Fluge and Mella also found disturbed amino acid metabolism.

Discussed here:
Metabolic profiling indicates impaired pyruvate dehydrogenase function in myalgic encephalopathy/CFS (Fluge et al., 2016)
https://forums.phoenixrising.me/thr...function-in-myalgic-encephalopathy-cfs.48446/
Under normal circumstances, human cells utilize carbohydrates, fats (lipids) and proteins (amino acids) as sources of energy, through catabolic processes in the mitochondria, the “powerhouses” of the cell.

[...]

The enzyme pyruvate dehydrogenase (PDH) plays an important role in the regulation of these processes, as it contributes to coordinating the utilization of carbohydrates, amino acids and lipids (fats) as energy sources.

[...]

In previous international studies, reduced levels of certain specific amino acids in the blood of ME patients have been reported. In our new study, all 20 standard amino acids were analysed in the blood of 200 patients included in clinical trials as well as 100 healthy control subjects.

A specific reduction in amino acids which are catabolized independently of the PDH enzyme was observed.