WASF3 disrupts mitochondrial respiration and may mediate exercise intolerance in myalgic encephalomyelitis/chronic fatigue syndrome

[hapl808]

My guess is that there is at least a significant portion of pwME that have undiagnosed neuromuscular conditions

I certainly do, but haven't noticed improvements when I've added or removed TUDCA.

 

[SlamDancin]

I certainly do, but haven't noticed improvements when I've added or removed TUDCA.

I’ve been researching different forms of muscular dystrophy, I think there is a good chance this is the underlying source of my illness, and I’ve see that these conditions involve other factors, metabolic dysfunction, mitochondrial stress, alterations to signaling hubs like mTOR and AMPK, calcium handling problems and ER or SR stress is a result of these. TUDCA may not be strong enough on its own or maybe not addressing underlying enough issues.

 

[hapl808]

I’ve been researching different forms of muscular dystrophy, I think there is a good chance this is the underlying source of my illness, and I’ve see that these conditions involve other factors, metabolic dysfunction, mitochondrial stress, alterations to signaling hubs like mTOR and AMPK, calcium handling problems and ER or SR stress is a result of these. TUDCA may not be strong enough on its own or maybe not addressing underlying enough issues.

Interesting. I've wondered about MD, but my neuromuscular doctor seemed to think that the pain I felt from muscular breakdown was not indicative of MD. Who knows, not super impressed by her even though she was well respected for disorders like MD.

 

[Violeta]

Leukotrienes ( LTC4 and -D4 )enhanced the TNF-α-induced MMP-9 production via binding the cysLT1 receptor in human monocytes/macrophages. (leukotrienes is only one of the things that enhances MMP-9 production)

MMP9 (matrix malloproteinase) may lower mannose binding lectins.

Regarding MBL :

MBL generates ER Stress and is also a cause of Liver Injury.

https://twitter.com/i/web/status/1695776654981836976

And MBL deficiency augments ER stress.

MMP is an enzyme that break down extracellular tissue. If MMP 9 is over 332 ng/mL, then this is treated with a low amylose diet and high dose fish oil. (I started a low amylose diet several days ago to see if it would help chronic morning cough, and it is helping. Yesterday I started a seaweed supplement, besides helping lower MMP9, it will provide iodine, hopefully that will help raise my temperature.)

Vitamin D, 25(OH)D, and 1,25(OH)2D inhibit MMP-2 and MMP-9 production in response to IL-1β, and also inhibits activation of MMPs induced by trypsin.

Matrix Metalloproteinase Inhibitors (MMPIs) from Marine Natural Products: the Current Situation and Future Prospects​

(fucoidan)

https://www.mdpi.com/1660-3397/7/2/71#b32-marinedrugs-07-00071

 

[SlamDancin]

Interesting. I've wondered about MD, but my neuromuscular doctor seemed to think that the pain I felt from muscular breakdown was not indicative of MD. Who knows, not super impressed by her even though she was well respected for disorders like MD.

I have yet to see a doc in that specialty. Not in all my years of seeing docs did any of them say that it might be MD and I should see a neurosmuscular expert. Now that I’m aware and familiar with the condition I’m almost shocked no one ever suspected it. I almost died from a relatively minor car accident after I developed rhabdo. Had to be hospitalized for 5 days until creatine kinase came back to safe levels and that’s actually the exact biomarker for a specific MD that is known to cause rhabdo. Luckily I have been able to heal the muscles after like five years of PT. A few supplements look promising too. My ME/CFS symptoms are greatly greatly improved and even if I can never do heavy exercise it does feel like a blessing just not getting PEM and all that. I hope you continue to pursue it. These MDs often cause the exact same main symptoms, metabolic disturbances, myalgia and exercise intolerance.

 

[hapl808]

A few supplements look promising too

What supplements?

Sadly, I've pursued it for many, many years with no success. I had what looked like rhabdo after an injury (sprained ankle) while traveling, but by the time I got back and was tested, CK was normal. My muscles did slowly heal for awhile, but then pushing through and a bit of an allergic reaction severely dropped my function and I never recovered.

 

[Violeta]

Salubrinol information is very interesting.

This article about salubrinol gives some background on what might be causing the ER stress. Wouldn't you think it would be helpful to remove whatever is causing the ER stress?

https://www.oarsijournal.com/article/S1063-4584(13)00707-3/fulltext

Using C28/I2 human chondrocytes, we applied ER stress with tunicamycin and inflammatory distress with tumor necrosis factor α (TNFα).

Stress to the endoplasmic reticulum (ER) and inflammatory cytokines induce expression and activity of matrix metalloproteinase 13 (MMP13). Since a synthetic agent, salubrinal, is known to alleviate ER stress and attenuate nuclear factor kappa B (NFκB) signaling, we addressed a question whether upregulation of MMP13 by ER stress and cytokines is suppressed by administration of salubrinal.

Methods​

Using C28/I2 human chondrocytes, we applied ER stress with tunicamycin and inflammatory distress with tumor necrosis factor α (TNFα) and interleukin 1β (IL1β). RNA interference with siRNA specific to NFκB p65 (RelA) was employed to examine a potential involvement of NFκB signaling in salubrinal's action in regulation of MMP13. We also employed primary human chondrocytes and evaluated MMP13 activity.

Results​

The result showed that tunicamycin activated p38 mitogen-activated protein kinase (MAPK), while inflammatory cytokines activated p38 MAPK and NFκB. In both cases, salubrinal significantly reduced expression and activity of MMP13. Silencing NFκB reduced inflammatory cytokine-driven upregulation of MMP13 activity.

Conclusions​

The results demonstrate that salubrinal downregulates expression and activity MMP13 through p38 and NFκB signaling, suggesting its potential usage to treat degenerative diseases such as osteoarthritis.

So it might help to figure out what is causing the inflammation and dealing with that.

 

[Violeta]

NFkB and muscle wasting.

Nuclear Factor-kappa B Signaling in Skeletal Muscle Atrophy​

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2597184/

Emerging evidence suggests that nuclear factor-kappa B (NF-κB) is one of most important signaling pathways linked to the loss of skeletal muscle mass in various physiological and pathophysiological conditions. Activation of NF-κB in skeletal muscle leads to degradation of specific muscle proteins, induces inflammation and fibrosis, and blocks the regeneration of myofibers after injury/atrophy. Recent studies employing genetic mouse models have provided strong evidence that NF-κB can serve as an important molecular target for the prevention of skeletal muscle loss. In this article, we have outlined the current understanding regarding the role of NF-κB in skeletal muscle with particular reference to different models of muscle-wasting and the development of novel therapy.

 

[SlamDancin]

@hapl808 I’ll try to PM you about it

 

[datadragon]

NFkB and muscle wasting.Nuclear Factor-kappa B Signaling in Skeletal Muscle Atrophy
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2597184/

Zinc regulates NF-kB as well as iNos. The Zinc Finger Protein Gfi1 Controls TLR4-Mediated Inflammatory Response by Directly Antagonizing NF-κB Transcription Factor https://www.sciencedirect.com/science/article/pii/S0006497119477677
Zinc regulates iNOS-derived nitric oxide formation in endothelial cells. Zinc inhibits iNOS promoter activity. NF-kB silencing abolishes cytokine-induced iNOS expression. Zinc inhibits the transactivation activity of NFκB. https://www.sciencedirect.com/science/article/pii/S2213231714000834

Zinc regulates BH4. https://forums.phoenixrising.me/threads/is-there-bh4-supplement-medicine.22480/post-2452006

And MBL deficiency augments ER stress.

Human mannose-binding lectin (MBL) is encoded by the MBL2 gene and is a key player in innate immunity. Human mannose-binding lectin 2 is directly regulated by peroxisome proliferator-activated receptors via a peroxisome proliferator responsive element In this study, we showed that PPARα and PPARγ up-regulate the expression of human MBL2.... https://pubmed.ncbi.nlm.nih.gov/23711995/ This is just one part I had mentioned earlier in the thread.

Interestingly, Zinc is required for PPARs: https://www.sciencedirect.com/science/article/pii/S0022316623029346?via=ihub

PPAR-a and zinc also regulate ER Stress. https://forums.phoenixrising.me/thr...dges-che-lipkin-fiehn-2023.90957/post-2453306 PPAR-a is also lowered from homocysteine. PPAR-a may also restore peroxisomal function and plasmalogen levels. I mentioned PPARa activation such as by using Palmitoylethanolamide (PEA)) or the synthetic drug fenofibrate and combined with chelated zinc/zinc amino acids may be one method suggested by the research to start but I have been discussing others.
There are many forums of zinc supplementation. https://www.onedaymd.com/2021/03/types-zinc-supplementation-absorption.html

PPARa activation (in this case by using Palmitoylethanolamide (PEA)) engages Allopregnanolone biosynthesis and induced marked antidepressive- and antianxiety effects. These effects were mimicked by the PPAR-α synthetic agonists, fenofibrate and GW7647, and were prevented by PPAR-α deletion, PPAR-α antagonists, and neurosteroid-enzyme inhibitors. (showing ppar-a is what is giving the effect). The endocannabinoid and neurosteroid systems regulate emotions and stress responses. The naturally occuring neurosteroid progesterone metabolite allopregnanolone prevents the activation of pro-inflammatory proteins important for gene regulation, as well as the creation of cytokines, which are known to be involved in many different inflammatory conditions. Allopregnanolone also tamps down chemokines and cytokines, such as NFkB, HMGB1, MCP-1 and TNF-a, all of which are part of the immune system and involved in many different inflammatory diseases. Allopregnanolone substantially blocked, or inhibited, the LPS-induced TLR4 activation in macrophages, which are found in white blood cells and part of the immune system, including in the brain. In macrophages, the TLR4 protein is part of a proinflammatory response that leads to the production of pro-inflammatory cytokines. https://www.sciencedirect.com/science/article/pii/S0006322319300812

 

[SlamDancin]

Can we just assume everyone is already taking their zinc so we can move forward?

 

[Violeta]

Zinc regulates NF-kB as well as iNos. The Zinc Finger Protein Gfi1 Controls TLR4-Mediated Inflammatory Response by Directly Antagonizing NF-κB Transcription Factor https://www.sciencedirect.com/science/article/pii/S0006497119477677
Zinc regulates iNOS-derived nitric oxide formation in endothelial cells. Zinc inhibits iNOS promoter activity. NF-kB silencing abolishes cytokine-induced iNOS expression. Zinc inhibits the transactivation activity of NFκB. https://www.sciencedirect.com/science/article/pii/S2213231714000834

Zinc regulates BH4. https://forums.phoenixrising.me/threads/is-there-bh4-supplement-medicine.22480/post-2452006


Human mannose-binding lectin (MBL) is encoded by the MBL2 gene and is a key player in innate immunity. Human mannose-binding lectin 2 is directly regulated by peroxisome proliferator-activated receptors via a peroxisome proliferator responsive element In this study, we showed that PPARα and PPARγ up-regulate the expression of human MBL2.... https://pubmed.ncbi.nlm.nih.gov/23711995/ This is just one part I had mentioned earlier in the thread.

Interestingly, Zinc is required for PPARs: https://www.sciencedirect.com/science/article/pii/S0022316623029346?via=ihub

PPAR-a and zinc also regulate ER Stress. https://forums.phoenixrising.me/thr...dges-che-lipkin-fiehn-2023.90957/post-2453306 PPAR-a is also lowered from homocysteine. PPAR-a may also restore peroxisomal function and plasmalogen levels. I mentioned PPARa activation such as by using Palmitoylethanolamide (PEA)) or the synthetic drug fenofibrate and combined with chelated zinc/zinc amino acids may be one method suggested by the research to start but I have been discussing others.
There are many forums of zinc supplementation. https://www.onedaymd.com/2021/03/types-zinc-supplementation-absorption.html

PPARa activation (in this case by using Palmitoylethanolamide (PEA)) engages Allopregnanolone biosynthesis and induced marked antidepressive- and antianxiety effects. These effects were mimicked by the PPAR-α synthetic agonists, fenofibrate and GW7647, and were prevented by PPAR-α deletion, PPAR-α antagonists, and neurosteroid-enzyme inhibitors. (showing ppar-a is what is giving the effect). The endocannabinoid and neurosteroid systems regulate emotions and stress responses. The naturally occuring neurosteroid progesterone metabolite allopregnanolone prevents the activation of pro-inflammatory proteins important for gene regulation, as well as the creation of cytokines, which are known to be involved in many different inflammatory conditions. Allopregnanolone also tamps down chemokines and cytokines, such as NFkB, HMGB1, MCP-1 and TNF-a, all of which are part of the immune system and involved in many different inflammatory diseases. Allopregnanolone substantially blocked, or inhibited, the LPS-induced TLR4 activation in macrophages, which are found in white blood cells and part of the immune system, including in the brain. In macrophages, the TLR4 protein is part of a proinflammatory response that leads to the production of pro-inflammatory cytokines. https://www.sciencedirect.com/science/article/pii/S0006322319300812

I didn't know zinc is required for PPAR and regulates BH4, too. Or that this would affect TLR4.

I'm starting to feel as though I have most of the bases covered. Just have to keep up with it all for the fatigue to reduce. Usually I get a bit more energy in the beginning of spring, which is just around the corner.

 

[Violeta]

Can we just assume everyone is already taking their zinc so we can move forward?

I learn a lot from the messages. You can skip over them.

 

[SlamDancin]

It’s the same post over and over and over, literally word for word

 

[Murph]

This paper (wasf3 disrupts ...) stands out in the me/cfs field because it proceeds from observation first. There's lots of work that proceeds from theory, for example the itaconate shunt. And lots of researchers like scheibenbogen who have observations but also a bunch of theory papers that race far ahead of what is actually confirmed.

I love this paper because it is very much just the facts. The quality of the work is really high (in part because it's using cancer research budget!)

Usually I wouldn't change my views based on one paper but this one inclines me to think that ER stress is a big part of the picture for us. I am increasingly suspicious that PEM is either the onset of the unfolded protein response. Or perhaps the start of PEM correlates with the point where the cell decides enough unfolded protein response (UPR) is enough and proceeds to apoptosis / necrosis.

However, if ER stress is crucial to me/cfs then we need to grapple with the biology of estrogen. Here are two papers that seem to matter. This first one seems to show that estrogen reduces ER sress markers.

Estrogen reduces endoplasmic reticulum stress to protect against glucotoxicity induced-pancreatic β-cell death ​

DOI: 10.1016/j.jsbmb.2013.09.018

Abstract ​

Estrogen can improve glucose homeostasis not only in diabetic rodents but also in humans. However, the molecular mechanism by which estrogen prevents pancreatic β-cell death remains unclear. To investigate this issue, INS-1 cells, a rat insulinoma cell line, were cultured in medium with either 11.1mM or 40mM glucose in the presence or the absence of estrogen. Estrogen significantly reduced apoptotic β-cell death by decreasing nitrogen-induced oxidative stress and the expression of the ER stress markers GRP 78, ATF6, P-PERK, PERK, uXBP1, sXBP1, and CHOP in INS-1 cells after prolonged culture in medium with 40mM glucose. In contrast, estrogen increased the expression of survival proteins, including sarco/endoplasmic reticulum Ca(2+) ATPase (SERCA-2), Bcl-2, and P-p38, in INS-1 cells after prolonged culture in medium with 40mM glucose. The cytoprotective effect of estrogen was attenuated by addition of the estrogen receptor (ERα and ERβ) antagonist ICI 182,780 and the estrogen membrane receptor inhibitor G15. We showed that estrogen decreases not only oxidative stress but also ER stress to protect against 40mM glucose-induced pancreatic β-cell death.

But this second one seems to show estrogen turning on something called the "anticipatory UPR. "
I'm not yet across what the anticipatory UPR is but the name alone is raising a big red flag for me!

A New Role for Estrogen Receptor α in Cell Proliferation and Cancer: Activating the Anticipatory Unfolded Protein Response​

Mara Livezey,1 Ji Eun Kim,1 and David J. Shapiro1,2,*
Author information Article notes Copyright and License information PMC Disclaimer

Go to:

Abstract​

Cells react to a variety of stresses, including accumulation of unfolded or misfolded protein, by activating the endoplasmic reticulum (EnR) stress sensor, the unfolded protein response (UPR). The UPR is highly conserved and plays a key role in the maintenance of protein folding quality control and homeostasis. In contrast to the classical reactive mode of UPR activation, recent studies describe a hormone-activated anticipatory UPR. In this pathway, mitogenic hormones, such as estrogen (E2), epidermal growth factor, and vascular endothelial growth factor rapidly activate the UPR in anticipation of a future need for increased protein folding capacity upon cell proliferation. Here, we focus on this recently unveiled pathway of E2-estrogen receptor α (ERα) action. Notably, rapid activation of the anticipatory UPR pathway is essential for subsequent activation of the E2-ERα regulated transcription program. Moreover, activation of the UPR at diagnosis is a powerful prognostic marker in ERα positive breast cancer. Furthermore, in cells containing ERα mutations that confer estrogen independence and are common in metastatic breast cancer, the UPR is constitutively activated and linked to antiestrogen resistance. Lethal ERα-dependent hyperactivation of the anticipatory UPR represents a promising therapeutic approach exploited by a new class of small molecule ERα biomodulator.

Once again here's the link to the original paper we are discussing in this thread which implicates endoplasmic reticulum stress, the unfolded protein response and a particular protein called WASF3 in me/cfs. It is very much worth reading:
https://www.pnas.org/doi/10.1073/pnas.2302738120

 

[SlamDancin]

@Murph I think also probably relevant is the sarcoplasmic reticulum, SR.

The SR is a specialized extension of the ER and coordinates the release of calcium during muscle contraction [7].

It’s also vulnerable to stress and SR stress is thought to contribute to the pathogenesis of skeletal muscle disorders

 

[mitoMAN]

I dont fully understand why you guys are so focused on TUDCA when NaPB is a critical part of the treatment.
TUDCA alone wont do the job, thats pretty obvious as too many have already tried that as a regular over the counter supplement.

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7904702/#:~:text=Sodium 4-phenylbutyrate (4-,critical role in neurological diseases.
Sodium 4-phenylbutyrate (4-PBA) reportedly acts as a chemical chaperone that alleviates endoplasmic reticulum (ER) stress, which plays a critical role in neurological diseases

The data showed that 4-PBA significantly reduced salinomycin-triggered ER stress
https://bmccancer.biomedcentral.com/articles/10.1186/s12885-019-5590-8

Thats why the planned clinical trial for LC will use Relyvrio which is NaPB + TUDCA for ALS
https://www.relyvrio.com/RELYVRIO-Directions-For-Use.pdf

 

[SlamDancin]

@mitoMAN it was discussed here I think, NaSB takes huge doses to overcome terrible bioavailability, if I recall correctly. I would be interested in 4-PBA potentially or Salubrinol

 

[mitoMAN]

@mitoMAN it was discussed here I think, NaSB takes huge doses to overcome terrible bioavailability, if I recall correctly. I would be interested in 4-PBA potentially or Salubrinol

4-PBA is another name for NaPB.
And yea dosage is up to 6g - which is affordable still (see my old posts.)
Salubrinol should be toxic and avoided

 

[Murph]

I dont fully understand why you guys are so focused on TUDCA when NaPB is a critical part of the treatment.
TUDCA alone wont do the job, thats pretty obvious as too many have already tried that as a regular over the counter supplement.

We all have to balance effectivness and accessibility. Sodium phenylbutyrate is undoubtedly likely to be superior at fixing ER stress. But getting it is tricky and it's safety seems less clear. Whereas tudca *should* work at least a bit and is super cheap and easy to get. I'm not ruling out using sodium phenylbutyrate if the data becomes clearer.