Cause of EBV Fatigue found + Biomarkers (Mitochondria “Cellular Chronic Fatigue")

sometexan84

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Big news recently (Feb 2020) for anyone w/ EBV, and possibly HHV-6. Good News and Bad News.

Good News - The cause of fatigue from EBV has been identified. Also, there are biomarkers for testing. And these biomarkers likely cover more than just EBV in Chronic Fatigue.

Bad News - Biomarkers for testing have not yet been set up due to interruption from COVID, despite how simple the testing process is.

Here are the links. I've gone through them extensively and will add bullet summaries below the links, along w/ an email response from those that made the discovery.

An Isolated Complex V Inefficiency and Dysregulated Mitochondrial Function in Immortalized Lymphocytes from ME/CFS Patients

Cell-Based Blood Biomarkers for Myalgic Encephalomyelitis/Chronic Fatigue Syndrome

As you can see, there are 2 articles. The 2nd one they published promptly after the first to show how it can be tested and diagnosed.

Personal Notes I took for my own reference:
  • Mitochondria Dysfunction - Complex V Deficiency “Cellular Chronic Fatigue”
  • There are (5) protein Complexes that are used for converting oxygen and glucose into energy w/in mitochondria (I, II, III, IV, and V)
  • These Complexes aid in this energy conversion, resulting in the production of ATP (adenosine triphosphate)
  • New study found that “As a proportion of the basal oxygen consumption rate (OCR), the rate of ATP synthesis by Complex V was significantly reduced in ME/CFS lymphoblasts”
  • The above study used EBV infected cells
  • It found that CFS patients DO exhibit a mitochondrial deficiency in ATP generation
  • The deficiency is specifically from Complex V rather than a generalized reduction in all mitochondrial functions
  • ATP levels stay normal, despite the deficiency of ATP synthesis.
  • SAME GROUP OF AUSTRALIANS that did the first study, promptly had a follow up ready, showing biomarkers.
  • They found 3 biomarkers, which together are extremely accurate:
  • 1) lymphocyte death rate
  • 2) mitochondrial respiratory function
  • 3) TORC1 activity

Additional Notes (for you guys):
  • The difference in this study is that they found what other people couldn't. Lots of recent studies found this and that about possible Mito dysfunction, but couldn't figure it out. They finally did here, with the discovery of one small part of the energy production process in mitochondria, Complex V.
  • Even though they did this w/ EBV infected cells, I have a feeling the specific HHV-6 dysfunction will be found and confirmed now very soon (if it hasn't been already).
  • Similarly, I think the diagnostic biomarker testing is likely to apply to HHV-6, and possibly other non-EBV CFS conditions.
  • This discovery proves those with persistent EBV infections have fatigue and energy depletion from mitochondrial dysfunction.
  • This finding was very tricky to find in other studies, largely in part because ATP levels stay normal. Our bodies are apparently good at compensating, which makes it difficult for scientists to get to the root.
  • The main point of discovery was seen by ATP as % of basal oxygen consumption rate (OCR). So, like (ATP x 100) / basal OCR
1595464499254.png

  • This means that multiple recent studies are correct, even though some talk about different things that are causing fatigue and energy suck. So like, this mito dysfunction is a direct cause of energy loss. But the mito dysfunction here, would also validate other studies talking about automonic dysfunction and how that causes fatigue.
  • The 2nd paper talks about the biomarkers and they seem to be extremely accurate
    • 1) lymphocyte death rate
    • 2) mitochondrial respiratory function
    • 3) TORC1 activity
  • I haven't looked a lot into the 2nd or 3rd one. But the first one, lymphocyte death rate, involves freezing your cells. Then seeing how fast the cells die in culture. Or rather, how many (%) cells dies w/in 48 hrs.
  • 1595465178749.png
This is actually pretty easy to accomplish. But I couldn't find anything online about how to do it. So I reached out to the guys that did the study, and unfortunately got the following response...

Thanks for getting in touch and for your kind comments about our work. The cell death assay we used is widely used in a research setting but requires skilled operators. Prior to Covid-19 we were planning this year to determine which of a large number of different cell death assays would be more applicable to measuring the lymphocyte death rates for ME/CFS tests in a clinical setting. Unfortunately our lab has been pretty much closed and access to patients blocked since March because of Covid-19. It's frustrating of course, because we would like to be able to get this out to patients as soon as practicable, as we think there is a great need.

To my knowledge we are the only people who have done this assay in relation to ME/CFS, so it is not clinically available nor clinically standardized and validated across different labs yet. I am also not aware of it being done routinely in pathology labs for other clinical tests....so at the moment I don't think there is anywhere I could recommend.

I am sorry I can't help more at this time.

Best wishes,
Paul.

Emeritus Professor of Microbiology,
La Trobe University,
VIC 3086,
AUSTRALIA.
 
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Wishful

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Impressive work! I hope it pans out and becomes a valid diagnostic test. It could also lead to treatments, or at least a better understanding of ME, which could lead to treatments.

Yay Australia! :thumbsup: (I've noticed a lot of good, innovative research papers from Australia. Maybe it's from being upside down all the time. :D)
 

Hip

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Interesting paper, I have not seen this discussed before on ME/CFS forums. The paper says:
We found that mitochondrial function in ME/CFS lymphoblasts is indeed abnormal, with an isolated Complex V deficiency accompanied by elevated capacity of Complexes I to IV, decreased membrane potential, upregulation of TORC1 activity and elevated expression of diverse mitochondrial proteins involved ATP-generating catabolic pathways.

So they found that in ME/CFS complex V was down, and complex I, II, III and IV were up. This seems at odds with the Cara Tomas 2019 paper which found complex I, II and IV the same in ME/CFS and controls.


As for the upregulation of TORC1 activity, one way to combat that is the drug rapamycin (sirolimus):
The activity of TORC1 can be blocked by Rapamycin via an indirect mechanism. In this case, rapamycin forms an inhibitory complex by binding to the TOR-associated immunophilin FKBP12 (FK506 binding protein 12 kDa) [8,9]. TORC2 is rapamycin-insensitive.
Source: here


Several ME/CFS patients noted substantial improvements in their ME/CFS from taking rapamycin — see this post.
 

junkcrap50

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Great find! Published February 2020 - easy to miss with COVID peaking.

Any clues as to possible treatments?
 
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Wishful

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There are two newer generations of rapamycin drugs; I'm not sure about availability though. Also, resveratrol, curcumin and caffeine inhibit TORC1 (not sure how effectively at normal dosage). Resveratrol and curcumin just make me feel worse. :grumpy:
 

sometexan84

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So they found that in ME/CFS complex V was down, and complex I, II, III and IV were up. This seems at odds with the Cara Tomas 2019 paper which found complex I, II and IV the same in ME/CFS and controls.
No dude. It only reiterates these new findings. In fact that Cara Tomas paper suggests that what they found in the new article might be the case. They were actually onto it already...

The lack of difference in complex activity in CFS PBMCs suggests that the previously observed mitochondrial dysfunction in whole PBMCs is due to causes upstream of the mitochondrial respiratory chain.

The measurements were completely different too. That's part of the key here. Also, the 2019 study was really looking at ATP production. That was what I was trying to talk about above, like that's the thing, the ATP synthesis doesn't end up changing that much.
 
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Hip

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In fact that Cara Tomas paper suggests that what they found in the new article might be the case. They were actually onto it already.

The Tomas paper seems to conclude that there is no difference between ME/CFS patients and healthy controls when it comes to mitochondrial complexes. So Tomas et al conclude the mitochondrial dysfunction must be upstream of the mitochondrial respiratory chain:
The lack of difference in complex activity in CFS PBMCs suggests that the previously observed mitochondrial dysfunction in whole PBMCs is due to causes upstream of the mitochondrial respiratory chain.



But these ME/CFS energy metabolism papers often contradict each other. An earlier Tomas paper contradicted the Myhill energy metabolism papers. And the Lawson study found higher than normal ATP levels the cells of ME/CFS patients.
 

WantedAlive

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The deficiency is specifically from Complex V

Fisher revealed this in the Emerge Conference last year. The deficiency is when under stress if I recall, not at rest.

I found in sickle cell disease a complex V inhibition induced by hemolysis which contributes to platelet activation, and I've wondered if that might similarly be the cause of ME/CFS sticky blood. Study published here.

I'm also rying to find a study I discovered in Alzheimers experiencing upregulated complexes I - III after exposure to EV secretion from neighbouring cells following TNF-alpha exposure I think, but with lowered ATP and proton leak it suggested a similar dysfunction to Fisher's study. I'll post if I find it.
 

WantedAlive

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Here's the video of Fisher's presentation at Emerge Conference March last year which might be easier to understand. He talks about the complex V impairment, and how he describes complexes I-IV are upregulated to compensate.

I discovered an Alzheimers study that might show a similar dysfunction but sadly they never studied complex V (although I read a complex V impairment exists in AD). In this study, they tested extracellular vesicles secreted in response to cytokine exposure (TNF-alpha). They found significant increases in mitochondrial oxygen consumption in naive cells exposed to these EV's.

As quoted:
This unexpected increase in mitochondrial activity may actually be a signal of mitochondrial stress, as we concomitantly observed dose-dependent increases in proton leak, suggesting that although oxidative phosphorylation is increased, the mitochondria are not actually functioning optimally. Indeed, a disproportionately large rate of mitochondrial oxygen consumption likely indicates an attempt to maintain ψm as proton leak across occurs
. It was reported there was no change in expression of complexes I-IV despite decline in OXPHOS, does this suggest a similar dysfunction to Fisher's finding maybe?
 

Wishful

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That was what I was trying to talk about above, like that's the thing, the ATP synthesis doesn't end up changing that much.

That's the impression I got from the explanation of the electron transport chain in "The Vital Question": that the ATP production could remain the same at normal operating levels, but the point at which it completely breaks down changes. Measuring an engine's exhaust while it's idling doesn't tell you that it has a problem that is going to blow its pistons when you stomp on the accelerator. All those conclusions from previous studies of mitochondrial function and ATP levels should be reviewed for what operating conditions they were done under.
 

Wishful

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One immediate test to do: see whether the 'something in the blood' will cause the same markers in cells from a healthy donor.

Another test would be to see if a TORC1 inhibitor (or enhancer) affects PWME. Actually, the paper probably suggests a number of similar tests that could be done.
 

Wishful

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the discovery of one small part of the energy production process in mitochondria, Complex V.

Just to say, Complex V is NOT a small part of the process, any more than any part of a pipeline is less important. If Complex V (also known as ATP Synthase) doesn't do its function properly, the earlier parts of the chain generate ROS, which either signals the mitochondria to adapt to the new conditions, or it signals the cell to kill itself.

ATP synthase is worth looking up. The head of the molecule spins at 21,000 rpm!
 

Hip

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I cannot seem to find any supplements or drugs which boost complex V.

There are quite a few substances which boost complexes I to IV, detailed in this post.
 

wabi-sabi

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So they found that in ME/CFS complex V was down, and complex I, II, III and IV were up. This seems at odds with the Cara Tomas 2019 paper which found complex I, II and IV the same in ME/CFS and controls.
From what I can understand, these papers, while both showing a problem in the mitos, show different problems. While I am always excited to see new research, i 'm just not sure what to make of it when results conflict with one another. Does anyone know of good review articles? Is anyone working to reconcile these results?
 

junkcrap50

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One immediate test to do: see whether the 'something in the blood' will cause the same markers in cells from a healthy donor.

Another test would be to see if a TORC1 inhibitor (or enhancer) affects PWME. Actually, the paper probably suggests a number of similar tests that could be done.
Bhupesh Prusty definitely needs to know about this paper. Someone should send it to him, preferably someone who can explain its importance in relation to his research.
 

Learner1

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The folks at MitoSwab told me the typical pattern for the folks they tested with a buccal swab method (correlated 855% with muscle biopsies) is low complex I and high complex IV.

I've fine 3 of their tests and found that this can be manipulated with nutrients, which I did after whacking my HHV6, CMV, and EBV with a year and a half of Valcyte.

I've normalized complexes I-III with significant nutrient interventions, including MitoQ, acetylcarnitine, manganese, B vitamins, BCAAsNMN, quercetin, melatonin, resveratrol, and phospholipids. However, complex IV is stubbornly high be at 427% of normal. @Hip none of your suggestions in yourvlinks say how to LOWER hyperactive complex IV.

PQQ and gingko did nothing.

Is there a clinical test for complex V?

And, my pyruvate and lactate are below range, as they are for several other patients I know. And, I seem to have a problem burning fatty acids, as evidenced by metabolites on an OAT, a NutrEval, and a treadmill metabolic test. Would love any insight on this...

I've compared notes and labs with other patients and find we are variations on a theme, not all identical. Seems like it's more useful once again to find the categories of patients and find treatments for the categories not a one size fits all solution.
 

junkcrap50

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I cannot seem to find any supplements or drugs which boost complex V.

There are quite a few substances which boost complexes I to IV, detailed in this post.
From a quick look last night, it looks like AICAR can boost ATP-Synthase (Complex V), but likely indirectly through upregulation of AMPK:
In the current study, we demonstrated that AMPK activation by AICAR significantly relieved UA-induced mitochondrial dysfunction, as demonstrated by increased mtDNA, mitochondrial complex I and V (ATP Synthase) activity, and ATP production and reduced mitochondrial ROS, MMP and UCP2 expression.

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6531502/

L6 myotubes treated for 16 h with AICAR presented a significant, ∼10-fold increase in PGC-1α and F1ATP synthase mRNA levels, and a 4-fold increase in citrate synthase mRNA. Co-treatment with the NOS inhibitor l-NAME significantly blunted the AICAR effect on both PGC-1α and F1ATP synthase mRNA expression, and completely prevented the AICAR effect on citrate synthase mRNA (Fig. 8)

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2988518/

Correlating with mitochondrial activity, AICAR also increased complex V (ATP synthase) activity in HUVECs transfected with wild-type or phosphomimetic forms . AICAR increased ATP production and decreased ROS generation in wild-type or phosphomimetic forms of DNMT1-, RBBP7-, or HAT1-transfected HUVECs but not in DNMT1-S730A–, RBBP7-S314A ...

https://stke.sciencemag.org/content/10/464/eaaf7478.full
 

junkcrap50

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https://pubmed.ncbi.nlm.nih.gov/21715656/

A yeast-based assay identifies drugs active against human mitochondrial disorders

Elodie Couplan 1 , Raeka S Aiyar, Roza Kucharczyk, Anna Kabala, Nahia Ezkurdia, Julien Gagneur, Robert P St Onge, Bénédicte Salin, Flavie Soubigou, Marie Le Cann, Lars M Steinmetz, Jean-Paul di Rago, Marc Blondel

Abstract

Due to the lack of relevant animal models, development of effective treatments for human mitochondrial diseases has been limited. Here we establish a rapid, yeast-based assay to screen for drugs active against human inherited mitochondrial diseases affecting ATP synthase, in particular NARP (neuropathy, ataxia, and retinitis pigmentosa) syndrome. This method is based on the conservation of mitochondrial function from yeast to human, on the unique ability of yeast to survive without production of ATP by oxidative phosphorylation, and on the amenability of the yeast mitochondrial genome to site-directed mutagenesis. Our method identifies chlorhexidine by screening a chemical library and oleate through a candidate approach. We show that these molecules rescue a number of phenotypes resulting from mutations affecting ATP synthase in yeast. These compounds are also active on human cybrid cells derived from NARP patients. These results validate our method as an effective high-throughput screening approach to identify drugs active in the treatment of human ATP synthase disorders and suggest that this type of method could be applied to other mitochondrial diseases.