An Isolated Complex V Inefficiency and Dysregulated Mitochondrial Function in Immortalized Lymphocytes from ME/CFS patients 9/2019

[Inara]

The third possibility, that Complex V is inefficient in ME/CFS cells because of the elevated use of the proton motive force by other processes, is suggested by the elevated proton leak we measured in ME/CFS lymphocytes

Would elevated proton leak explain burning muscles?

 

[jaybee00]

@HTester

Your thoughts on the Fisher study?

Thanks

 

[nandixon]

I'm reposting a post below which I posted on another thread because this is where it actually belongs and because I'm going to be following up shortly when I have more energy about the problem I noted in the "Edit 2" at the bottom of the post.

(That upregulation of ATP5B that Dr Fisher found actually makes a microRNA a less likely candidate to be the 'something in the blood,' and I've found now where a particular protein has been shown to have all of the very same effects on mitochondrial function that Fisher found, and it has also been found to be carried in exosomes, but I'll mention more about all of this soon.)

I'm just putting this out very quickly as it is quite late. There is a very interesting and potentially very exciting connection between Paul Fisher's work[1] showing an impairment in ATP synthase in ME/CFS immortalized lymphocytes and Bhupesh Prusty's work[2] showing increased fission (and decreased fusion) in mitochondria treated with ME/CFS serum, possibly due to HHV-6A (and potentially other herpes viruses like EBV and CMV).

This connection is telling us that the 'something in the blood' may be targeting ATP synthase. In particular, that it may be targeting and down-regulating a subunit of ATP synthase known as ATP5B.

It turns out that ATP5B regulates mitochondrial fission and fusion in mammalian cells.[3]

If it is the ATP5B subunit being specifically targeted then that makes the 'something in the blood' more likely to be a microRNA (perhaps being carried by an exosome) versus a peptide, for example, which might be more likely if other parts of ATP synthase were being targeted.

Prusty did find a microRNA that HHV-6A promotes the expression of and that causes mitochondria to fission.[2] This microRNA may be targeting ATP5B.

(There is at least one compound, a small molecule known as M1 that may be capable of reversing such a down-regulation of ATP5B.[4])

[1] https://forums.phoenixrising.me/thr...lized-lymphocytes-from-me-cfs-patients.77577/

[2] https://www.healthrising.org/blog/2019/04/15/nih-chronic-fatigue-lipkin-davis-prusty-oh/

[3] https://www.tandfonline.com/doi/full/10.1080/19768354.2016.1188855

[4] https://www.ncbi.nlm.nih.gov/pubmed/22907892

Edit: I emailed Dr Fisher to take a look at this possibility.

Edit 2: Hmm, I just took another look at Dr Fisher's paper and he had found ATP5B to be upregulated. Not sure if that can be reconciled or not.

 

[godlovesatrier]

Interesting. I noticed 132g to 144g of protein per day to be very helpful for me especially. Although finding alternative forms of protein once food boredom sets in is tricky!

But it's interesting about the mTor - that sort of would make sense in terms of needing more of everything.

 

[nandixon]

Just cross-posing the following, since the Ciregia study supports Fisher's findings:

The findings of the 2016 Ciregia study (full text here:
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5048217/) have now been essentially validated to a fairly good extent with respect to ATPB, which is also known as ATP5B, a subunit of ATP synthase (aka Complex V).

In Paul Fisher's recent study of this thread:

An Isolated Complex V Inefficiency and Dysregulated Mitochondrial Function in Immortalized Lymphocytes from ME/CFS Patients
https://forums.phoenixrising.me/thr...lized-lymphocytes-from-me-cfs-patients.77577/

It was also found that the level of ATP5B protein was increased in immortalized lymphocytes from ME/CFS patients:

View attachment 35006

The p-value was not impressive at 0.049, but related findings in Fisher's study have excellent p-values and strongly suggest that ATP5B is indeed upregulated as Ciregia found.

I have a hypothesis that the "something in the blood" that Ron Davis and several other groups have noted may be targeting ATP5B and I may have found an interesting specific candidate for what that something is. I'll be posting more about this on the Fisher thread fairly soon. (I'm still working on trying to disprove it.)

 

[S-VV]

Do you believe the upregulation of ATP5B is compensatory or pathological?

 

[wigglethemouse]

I have a hypothesis that the "something in the blood" that Ron Davis and several other groups have noted may be targeting ATP5B and I may have found an interesting specific candidate for what that something is.

Ooooooooooh. Can you give us a clue?

The only thing I could find/guess at (using search function on PR) is that at one time there was some thought that measurement antibodies used to measure Viral protein VP1 for enterovirus detection (perhaps also common cold virus) can cross react with mitochondrial ATP5B and CKB proteins. And after watching the Prusty video about HHV1-8 sequencing having a common sequence gave me the idea that possibly ATP5B and VP1 have a common sequence that antibodies are targeting?
https://www.ncbi.nlm.nih.gov/pubmed/24190581

I've no idea on how to look into that. And if it was the case, how could such a theory explain persistence in immortalized lympocytes/lymphoblasts.

 

[wigglethemouse]

I have a hypothesis that the "something in the blood" that Ron Davis and several other groups have noted may be targeting ATP5B and I may have found an interesting specific candidate for what that something is. I'll be posting more about this on the Fisher thread fairly soon. (I'm still working on trying to disprove it.)

@nandixon
Just a suggestion in case it helps in working through your idea - @mariovitali is very generous with his time and custom machine learning tools he has built to investigate ME and would like to see them put to good use. If you would like his help to try and find more supporting information to prove or disprove your idea on the possible cause of the Complex V issue he can enter your search terms into his tools and output a list of relevant topics and papers............

 

[mariovitali]

@nandixon @wigglethemouse

Using NER (Named-entity recognition) i first identify what kind of Genes are found on all PUBMED papers mentioning ATP5B .Then i cross-referenced things i found with ATP5B. These are :

1) PPARGC1A aka PGC-1α

https://www.ncbi.nlm.nih.gov/pubmed/?term=ATP5B+PPARGC1A




Association of PGC-1a with ATP5B

These findings suggest that, in addition to promoter regions, a large proportion of PPARGC1A binding events occur at distal regulatory elements that may correspond with enhancer regions. Importantly, PPARGC1A peaks were found adjacent to numerous previously identified target genes, including PCK1, ALAS1, ACADM, ATP5B, CYP7A1, CYP8B1, TRIB3, CYCS, ESRRA, and SOD2, further supporting the validity of our binding data set (Supplemental Table S2). Gene ontology (GO) analysis (The Gene Ontology Consortium 2000) of PPARGC1A-occupied genes revealed enrichment of several functional categories that have been associated with PPARGC1A, such as oxidative phosphorylation and lipid metabolism, in addition to novel categories such as protein folding

https://genome.cshlp.org/content/genome/22/9/1668.full.html

Interestingly, ALAS1, CYP7A1 and CYP8B1 are related to bile acids.

2) ATP5B and Glutamate Dehydrogenase (GDH)

Two additional proteins that co-purify with mGluR1b, ATP synthase beta chain (ATP5B) and glutamate dehydrogenase, are metabolic enzymes present in the inner mitochondria matrix. The presence of mitochondrial proteins in the mGluR1b complex may possibly be due to non-specific interactions formed during the purification process and not to interactions formed in intact cells. However, both ATP synthase beta chain (Chi and Pizzo 2006) and glutamate dehydrogenase are also present in subcellular compartments outside of mitochondria. ATP5B localizes to the cell membrane where it binds apolipoprotein A–I (Martinez et al. 2003). In addition, ATP5B binds to caveolin-1 and translocates to plasma membrane caveolae in response to cholesterol (Wang et al. 2006). Membrane-bound glutamate dehydrogenase has been shown to be involved in the association of lysosomes to microtubules, to which it binds in an ATP-dependent manner (Rajas et al. 1996). At present, the specific function/s of ATP5B and glutamate dehydrogenase outside of mitochondria are not well understood.

3) ATP5B and Glutamate (Mice).

The subsequent iTRAQ experiment identified 2242 proteins from the ischemic hemisphere with <1.0% false discovery rate. Data mining revealed that (1) about 2.7% of detected proteins were temporally perturbed having an involvement in the energy metabolism (Pygb, Atp5b), glutamate excitotoxicity (Slc1a3, Glud1), neuro-inflammation (Tf, C3, Alb), and cerebral plasticity (Gfap, Vim, Gap43);

https://www.ncbi.nlm.nih.gov/pubmed/21950801


and more on ATP5B and GDH

https://www.cell.com/cell-reports/pdfExtended/S2211-1247(18)31268-3


and ATP5B and excitotoxicity


http://www.rki-i.com/cell regulation 2018/acel12715.pdf


Hope this helps !

 

[nandixon]

@wigglethemouse, I'm hoping to post something fairly soon. I've been very short of extra energy the last few weeks from having to exert myself physically much more than usual due to a drought on my property and moreover because of some eye surgery that went wrong and that won't get fixed for 2 more weeks. I may just post my idea for the "something in the blood" in a sort of bullet point format and not try to do as much explaining as I might otherwise. I'm having a lot of problems seeing well though and it's exhausting.

 

[wigglethemouse]

I'm having a lot of problems seeing well though and it's exhausting.

Oh shoot. Take care and you can update the thread when you feel able.

 

[wigglethemouse]

@nandixon Here is the recent University of Otago, New Zealand, talk where the researcher talks about their 10 patient Seahorse, proteomics, transcriptomics study (copy and paste link for talk).

https://www.youtube.com/watch?v=vlDGEckRNyk&feature=youtu.be

At 9mins she claims the proteomics results replicate Paul Fishers findings ! Here is the slide

 

[bread.]

If it was important, the OMF and other players would jump on it, no?

@HTester @Ben H
@Janet Dafoe (Rose49)

Saying that, I am still certain ME is a mitochondrial disease primarily and that is not a good thing I am afraid.

 

[percyval577]

Saying that, I am still certain ME is a mitochondrial disease primarily and that is not a good thing I am afraid.

Though an intrinsic one has become a bit unlikely now:

threads/mtdna-population-variation-in-myalgic-encephalomyelitis-chronic-fatigue-syndrome-in-two-populations-a-study-of-mildly-deleterious-variants.78332/ Venter et al 2019, in fact in this view the mt even work better.

Also Schoeman 2017 in the literature, I don´t know what Gorman 2015 says, and what´s about Billing-Ross 2016.

There is finally another study with permealised Mt, and all works fine, in itself (link in the thread).

 

[bread.]

mtdna mutations or the lack thereof says potentially absolutely nothing about the functioning of any given patients mitochondria, neither is there one single test or a a multitude of tests, not even in research facilities only, that can give you a good picture of your mitochondria, it is somewhat of a black box at this point.

There are people with mitochondrial disease that have a clear cut diagnosis due to available testing, those people very often have the same symptoms as many ME patients, especially the severe ones.

All theses diagnoses like POTS, SFNP, Dysautonomia and MCAD are extremely common in all sorts of identifiable mitochondrial disease, so are they in ME.

We know that we know very little about mitochondria and have very limited resources to diagnose diseases that are caused by faulty ones that are not genetically malfunctioning (but in all likelihood we don‘t know all genetically predisposed also).

We know about „damaged“ and „fused“ mitochondria according to Dr. Davis.

So I think, no 100% proof of ME being a mitochondrial disease means very little.

Also, not all ME/CFS patients will have mitochondrial disease, because there is no good subtyping in ME/CFS in studies at this point it will be difficult to find anything conclusive.

 

[SlamDancin]

@bread The Lights at University of Utah have done studies of mitochondria genetic variants and for instance I have a possibly defunctional ATP7A gene. Certain cells need this protein to utilize copper in the mitochondria to make ATP. Specifically collagen producing cells need it and noradrenergic neurons. So my OI and structural issues do match up with this testing.

I also had a couple possibly deleterious SNP’s according to the group in mt-nd1 and mt-nd2 and also NQO1 (part of complex 1).

 

[percyval577]

Specifically collagen producing cells need it and noradrenergic neurons.

I too seem to have a collagen issue (at least visible so the two times when improving, paradoxically), and an issue with NA.

Though you might extent and shift a bit your imagination: https://forums.phoenixrising.me/threads/mitochondra-in-me-cfs.78406/

 

[mariovitali]

@nandixon Here is the recent University of Otago, New Zealand, talk where the researcher talks about their 10 patient Seahorse, proteomics, transcriptomics study (copy and paste link for talk).

https://www.youtube.com/watch?v=vlDGEckRNyk&feature=youtu.be

At 9mins she claims the proteomics results replicate Paul Fishers findings ! Here is the slide
View attachment 35503

From the same presentation, something that may be interesting :

There are patients with ME that have liver issues. I believe that this aspect has not been investigated enough.

I sent an e-mail to Prof Tate but didn't get back a reply.

 

[wigglethemouse]

This is the final version of the paper published by MDPI/International Journal of Molecular Sciences on 6 Feb 2020
https://www.mdpi.com/1422-0067/21/3/1074

 

[msf]

I'm reposting a post below which I posted on another thread because this is where it actually belongs and because I'm going to be following up shortly when I have more energy about the problem I noted in the "Edit 2" at the bottom of the post.

(That upregulation of ATP5B that Dr Fisher found actually makes a microRNA a less likely candidate to be the 'something in the blood,' and I've found now where a particular protein has been shown to have all of the very same effects on mitochondrial function that Fisher found, and it has also been found to be carried in exosomes, but I'll mention more about all of this soon.)

I just read through the later posts on this thread, including this one, and thought I should post a link to this thread of mine here: https://forums.phoenixrising.me/thr...nergy-problem-in-me.80389/page-4#post-2293520

In short, based on this study and some other stuff I read, I tried Chinese skullcap as a treatment for PEM, with very impressive results (prevention of PEM to a large degree and the ability to exercise freely again). Baicalin (an ingredient in Chinese skullcap) has been shown to correct infection-induced decreases in ATPase: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6781171/