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

[Alvin2]

NaPB is a critical part of the treatment

Can't get it here in Canada.
By prescription you need special coverage which will only work if you have the specific diagnosed condition its indicated for (i forget which one, i had looked into it when that WASF3 paper came out).

 

[Forummember9922]

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.

Can I pester you for sources on the butyrate superiority? Will re add to the stack. Tudca was very non benign for me but made me kind of unstable on day 3-4. Of course couldve been other things, or a ‘herx’ of sorts that I didn't power through. Story of many of my trials.

 

[Murph]

Here's an interesting study: These guys got tired of there being a bunch of conflicting studies on UPR in Amyotrophic Lateral Sclerosis and set out to resolve them once and for all with a huge, powerful study done PROPERLY.​

It brings bad news: science doesn't really understand what the f is going on with the unfolded perotein response and/or its role in disease. ​

In the end all their results are kind of ??. They didn't find any anti-PERK drugs did anything for ALS. They also don't even find that Salubrinal does anything to stop the activaiton of UPR genes !! So if salubrinal was working on the cells of our patient with high WASF3, we're going to need to dig in on what was acually happening there.

This is great science. It says: These little studies you're basing your conclusions on aren't strong enough and we need to start again with our understanding of what is happening in this disease. It should hopefully stop people spending time and money exploring dead ends.

It's bad news for us though. Hwang can't piggyback on a great understanding of UPR in disease and how to treat it, because we don't seem to have one. He's going to need to do some basic science himself

PERK modulation, with GSK2606414, Sephin1 or salubrinal, failed to produce therapeutic benefits in the SOD1G93A mouse model of ALS​

• Fernando G. Vieira,
• Theo Hatzipetros

Published: February 15, 2024
https://doi.org/10.1371/journal.pone.0292190

 

[Murph]

Can I pester you for sources on the butyrate superiority? Will re add to the stack. Tudca was very non benign for me but made me kind of unstable on day 3-4. Of course couldve been other things, or a ‘herx’ of sorts that I didn't power through. Story of many of my trials.

Hwang is trying to do a trail of relyvrio which includes sodium phenylbutyrate (not sodium butyrate!). that's why we're guessing it's better.
https://www.s4me.info/threads/wasf3...ome-2023-hwang-et-al.34776/page-9#post-494479

 

[Murph]

Here's another treatment that people already attest to that fits with an andoplasmic reticulum stress model of me/cfs: vitamin d3

https://pubmed.ncbi.nlm.nih.gov/38352211/

1,25-dihydroxyvitamin D3 affects thapsigargin-induced endoplasmic reticulum stress in 3T3-L1 adipocytes​

Dain Wi 1 , Chan Yoon Park 1

Affiliations

• PMID: 38352211
• PMCID: PMC10861344
• DOI: 10.4162/nrp.2024.18.1.1

Free PMC article

Abstract​

Background/objectives: Endoplasmic reticulum (ER) stress in adipose tissue causes an inflammatory response and leads to metabolic diseases. However, the association between vitamin D and adipose ER stress remains poorly understood. In this study, we investigated whether 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) alleviates ER stress in adipocytes.

Materials/methods: 3T3-L1 cells were treated with different concentrations (i.e., 10-100 nM) of 1,25(OH)2D3 after or during differentiation (i.e., on day 0-7, 3-7, or 7). They were then incubated with thapsigargin (TG, 500 nM) for an additional 24 h to induce ER stress. Next, we measured the mRNA and protein levels of genes involved in unfold protein response (UPR) and adipogenesis using real-time polymerase chain reaction and western blotting and quantified the secreted protein levels of pro-inflammatory cytokines. Finally, the mRNA levels of UPR pathway genes were measured in adipocytes transfected with siRNA-targeting Vdr.

Results: Treatment with 1,25(OH)2D3 during various stages of adipocyte differentiation significantly inhibited ER stress induced by TG. In fully differentiated 3T3-L1 adipocytes, 1,25(OH)2D3 treatment suppressed mRNA levels of Ddit3, sXbp1, and Atf4 and decreased the secretion of monocyte chemoattractant protein-1, interleukin-6, and tumor necrosis factor-α. However, downregulation of the mRNA levels of Ddit3, sXbp1, and Atf4 following 1,25(OH)2D3 administration was not observed in Vdr-knockdown adipocytes. In addition, exposure of 3T3-L1 preadipocytes to 1,25(OH)2D3 inhibited transcription of Ddit3, sXbp1, Atf4, Bip, and Atf6 and reduced the p-alpha subunit of translation initiation factor 2 (eIF2α)/eIF2α and p-protein kinase RNA-like ER kinase (PERK)/PERK protein ratios. Furthermore, 1,25(OH)2D3 treatment before adipocyte differentiation reduced adipogenesis and the mRNA levels of adipogenic genes.

Conclusions: Our data suggest that 1,25(OH)2D3 prevents TG-induced ER stress and inflammatory responses in mature adipocytes by downregulating UPR signaling via binding with Vdr. In addition, the inhibition of adipogenesis by vitamin D may contribute to the reduction of ER stress in adipocytes.

 

[datadragon]

They didn't find any anti-PERK drugs did anything for ALS.

This study demonstrates that the activation of PPAR-α via the drug fenofibrate leads to neuroprotection by both reducing neuroinflammation and protecting mitochondria, which leads to a significant increase in survival in SOD1G93A mice. These mice develop ALS-like phenotypes from the age of 85–90 days and progressively weaken, lose weight, develop asymmetrical hind-limb tremor and paralysis, gait abnormalities, muscle atrophy, impaired motor performance, forelimb paralysis at end-stage and premature death around 128 days of age. Treated animals showed a significant slowing of the progression of disease with weight loss attenuation, enhanced motor performance, delayed onset and survival extension. Histopathological analysis of the spinal cords showed that neuronal loss was significantly attenuated in fenofibrate-treated mice. Mitochondria were preserved as indicated by Cytochrome c immunostaining in the spinal cord, which maybe partly due to increased expression of the PPAR-γ co-activator 1-α. The total mRNA analysis revealed that neuroprotective and anti-inflammatory genes were elevated, while neuroinflammatory genes were down-regulated. Therefore, the development of therapeutic strategies to activate PPAR-α as well as other PPARs may lead to new therapeutic agents to slow or halt the progression of amyotrophic lateral sclerosis. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4706116/

Zinc is needed for PPARs I mentioned a few posts up so I had 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.

Our data suggest that 1,25(OH)2D3 prevents TG-induced ER stress and inflammatory responses in mature adipocytes by downregulating UPR signaling via binding with Vdr. In addition, the inhibition of adipogenesis by vitamin D may contribute to the reduction of ER stress in adipocytes.

VDR signaling also requires Zinc and Vitamin A (RXR) for the VDR functions of Vitamin D to function properly. In other words some of those functions are regulated by zinc status. Zinc is required for the DNA binding proteins involved in the regulation of gene expression and is involved in Vitamin A metabolism. Vitamin D has other functions beyond VDR but these VDR functions would be impacted such as during prolonged inflammation/infection states.
https://forums.phoenixrising.me/threads/dang-those-vitamin-d3-levels.91152/post-2450071
https://forums.phoenixrising.me/threads/dang-those-vitamin-d3-levels.91152/post-2449906
https://forums.phoenixrising.me/thr...lightly-darker-anyone-else.91305/post-2450237

AGAIN, PPAR-a and zinc also regulate ER Stress and the UPR https://forums.phoenixrising.me/thr...s-chronic-fatigue-syndrome.90582/post-2453550

 

[datadragon]

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

The issue is more that in the presence of the pro inflammatory cytokines zinc is being sequestered in the cell and not just that its absorption is reduced. We can use chelated zinc/zinc amino acids to help at least with that part of it, as this may help overcome the zinc uptake/absorption issues and that is shown to work in studies, but I wasnt sure if it was enough alone to overcome zinc being brought into the cell and sequestered which was induced by the presence of the pro inflammatory cytokines. That is where the original idea to combine it with NLRP3 or ER Stress inhibitors that would lower the pro inflammatory cytokines at the same time and allow zinc to function as it does normally in a non inflammatory state. However, I found then it is also possible to RELEASE zinc from being sequestered in the cell. I already happened to have the research actually on how to release zinc from being sequestered.

I just reposted this but forgot to mention as to why we are sequestering zinc that you are missing: Pathogens must acquire trace minerals in order to replicate and cause disease. However, during infection, the host sequesters key nutrients restricting access to these nutrients as a part of a process known as nutritional immunity. Its part of the normal response to infection but can be problematic if prolonged or due to chronic inflammation which has the same response. Heres a small part: https://www.yeastinfectionadvisor.com/zinc.html
https://pubmed.ncbi.nlm.nih.gov/27242763/
https://www.cell.com/immunity/pdf/S1074-7613(13)00428-7.pdf When you understand all these relationships of zinc such as copper and iron, vitamin A, B6 and their involvement in various biological processes and pathways we can start to see how so many different symptoms or problems can crop up all at the same time.

I have posted one part of that if it helps you understand:
The cytokine interleukin 6 (IL6) induces the expression of Metallothionein and α2-macroglobulin (A2M) and consequently reduces zinc availability. IL-6 is released during the acute phase of an inflammatory response. This mechanism is beneficial to the acute immune response, however, a long-term decrease in zinc availability may contribute to pathological processes in conditions of chronic inflammation. Interleukin-6 (IL-6) up-regulates the ZIP14 gene expression, which in turn, is responsible for an excess of intracellular zinc and, at the same time, for hypozincemia that accompanies the acute phase response to inflammation and infection. Infection and inflammation produce systemic responses that include hypozincemia and hypoferremia. Interleukin-6 regulates the zinc transporter Zip14 in the liver and contributes to the hypozincemia of the acute-phase response. https://forums.phoenixrising.me/threads/unfolded-protein-response-and-a-possible-treatment-for-cfs.37244/post-2451850 Interferon-y (intensive exercise and high glucose also is ifn-y), Interferon-α (IFN-a) and inflammatory cytokines IL-1β, IL-6 and TNF-a, have all been shown to induce metallothioneins in the research, which can all reduce zinc availability and then further reduce its uptake/absorption in the gut

 

[Murph]

I just reposted this but forgot to mention as to why we are sequestering zinc

I appreciate you putting some context around why you're posting about zinc. it is hard for me to grasp the biological interactions that lead you to think zinc, of all vitamins and minerals, might be relevant. It never hurts to reiterate the full set of conceptual linkages that you are carrying in your mind for us, the readers.

 

[Violeta]

Hwang is trying to do a trail of relyvrio which includes sodium phenylbutyrate (not sodium butyrate!). that's why we're guessing it's better.
https://www.s4me.info/threads/wasf3-disrupts-mitochondrial-respiration-and-may-mediate-exercise-intolerance-in-myalgic-encephalomyelitis-chronic-fatigue-syndrome-2023-hwang-et-al.34776/page-9#post-4944f

Anyone have any thoughts about this study using sodium butyrate to induce EBV lytic reactivation?

"Accordingly, andrographolide inhibited the expression of Zta and viral production in sodium butyrate (NaB)-induced EBV lytic reactivation."

https://www.researchgate.net/public...the_Modulation_of_Epigenetic-Related_Proteins

 

[junkcrap50]

Anyone have any thoughts about this study using sodium butyrate to induce EBV lytic reactivation?

"Accordingly, andrographolide inhibited the expression of Zta and viral production in sodium butyrate (NaB)-induced EBV lytic reactivation."

https://www.researchgate.net/public...the_Modulation_of_Epigenetic-Related_Proteins

Yes. I have. See this article below that discusses it's use specifically to treat latent EBV infected T-Cells. Scroll down to "Antiviral Therapy" & "Targeting the EBV-infected T cells". (It's for a different disease, but same application could be used in CFS.) The paper also lists a couple other papers discussing this strategy for EBV. Their approach is to use a Histone deacetylase (HDAC) inhibitor + Ganciclovir.

The key and very cool part of this approach, I think, is that it hypothetically doesn't really allow much viral replication or any lysis, thereby containing the reactivated virus within the cell because of how ganciclovir works. (At least this is my understanding.) Ganciclovir causes cell death (I assume an autophagy process) when it is phosphorylated, which is done by EBV-protein kinsases, which are expressed/made by the reactivation inducer like sodium butyrate. Thus, you could preload you body saturating all your cells with ganciclovir inactivated, then give the HDAC inhibitor to activate the ganciclovir and trigger autophagy of all latent infected cells.

Of course, I'm not sure if it would practically work so straightforwardly. There is a risk of reactivating EBV, possibly causing spread, reinfection, or collateral damage. Potentially you could control the rate of EBV elimination by controlling the dosing of the reactivation inducer, but I'm not sure. But, you very well could make your CFS much worse by getting another EBV triggering of CFS. As a result, I'm not sure if you would be able to find a doctor willing to try this.

I'm wishfully hopeful this approach would work as I know two ME/CFS patients 100% cured getting the antiviral IV Vistide because they had high IgG EBV titers. So, essentially nuking any EBV in them cured them.

https://ashpublications.org/blood/a...ow-I-treat-T-cell-chronic-active-Epstein-Barr

Targeting the EBV-infected T cells​

For symptomatic patients (without HLH), we hypothesize that 1 potentially effective strategy includes combining ganciclovir, an antiviral agent, with drugs that can induce lytic gene expression in EBV-infected T cells (Figure 2). The latently infected EBV+ T cells are resistant to nucleoside-type antiviral agents because the drugs are not activated in these cells as the EBV-protein kinase (BGLF4) is not expressed.26 Histone deacetylase inhibitors such as vorinostat or romidepsin, proteasome inhibitors such as bortezomib, and short-chain fatty acids such as butyrate induce EBV protein kinase expression in latently infected cells, which phosphorylates ganciclovir, resulting in cell death. One example of such a strategy is using a schedule of bortezomib (velcade) and ganciclovir (or oral valganciclovir) as per a previous clinical trial targeting EBV+ lymphomas (registered at www.clinicaltrials.gov as #NCT00093704). Although there has been anecdotal success in our experience, there are as yet no comprehensive reports documenting the response rate and efficacy of this approach. Typically, patients receive bortezomib (1.3 mg/m2) IV over 3 to 5 seconds on days 1, 4, 8, and 11 and receive ganciclovir IV twice daily on days 1 to 14. We also propose that it is important to use ganciclovir and not acyclovir in this setting. Phosphorylated ganciclovir is toxic and kills infected cells, whereas phosphorylated acyclovir does not kill infected cells (although it does inhibit virus replication). Although oral valganciclovir can be used instead of ganciclovir, we recommend starting with ganciclovir first and then, when the viral load is more controlled, switching to the oral formulation.


Figure 2.

View largeDownload PPT
Treatment of EBV-infected lymphocytes with bortezomib or histone deacetylase (HDAC) inhibitors along with GCV can induce apoptosis of infected cells. EBV-infected T cells contain episomal EBV DNA (A). Bortezomib and HDAC inhibitors induce virus replication and expression of the EBV PK (B). GCV is phosphorylated by the viral PK, resulting in inhibition of EBV replication; phosphorylated GCV induces cell-cycle arrest and apoptosis of virus-infected cells (C). P, phosphate; PPP, triphosphate.

Treatment should be repeated every 21 days for a maximum of 3 courses. We recommend the following for hepatic impairment: for mild impairment (bilirubin ≤1 times upper limit of normal [ULN] and aspartate aminotransferase > UNL or bilirubin >1-1.5 times ULN), no initial dose adjustment required; for moderate (bilirubin >1.5-3 times ULN) or severe impairment (bilirubin >3 times ULN), reduce initial dose to 0.7 mg/m2 in the first cycle; based on patient tolerance, dose escalation to 1 mg/m2 or further dose reduction to 0.5 mg/m2 in subsequent cycles may be considered.

Similarly, arginine butyrate in combination with ganciclovir was administered to 15 patients with refractory EBV+ lymphoid malignancies of B-cell or NK/T-cell origin. Ganciclovir was administered twice daily at standard doses, and arginine butyrate was administered by continuous infusion in an intrapatient dose escalation, from 500 mg/kg per day to 2000 mg/kg per day, as tolerated, for a 21-day cycle. Ten of 15 patients showed significant antitumor responses, with 4 complete responders and 6 partial responder (PRs) within 1 treatment cycle. Among the complete responders, 1 patient had extranodal NK/T-cell lymphoma, and 1 patient had peripheral T-cell lymphoma. Among the partial responders, 1 patient had extranodal NK/T-cell lymphoma, and 1 patient had a subcutaneous panniculitis-like T-cell lymphoma. The combination of arginine butyrate and ganciclovir was felt to be reasonably well tolerated, with biologic activity in vivo.27 However, at present, the drug is not available for clinical use.

Therefore, another approach would to use the histone deacetylase inhibitor romidepsin, which is a potent inducer of EBV replication and in combination with ganciclovir has been shown to kill virus-infected cells in vitro and in a mouse model.28 Romidepsin also markedly induced EBV replication in patients and resulted in elevated liver enzymes and fever in NK/T-cell lymphoma.29 The drug is approved for treatment of cutaneous T-cell lymphoma. Hence, when administered with ganciclovir, romidepsin might be used as an alternative to bortezomib for T-cell CAEBV, although close clinical monitoring would be critical.

Other immunomodulatory/T-cell–directed strategies tried have included IFN-α, hydroxyurea, and lenalidomide, none of which have shown robust success.30 HSP90 inhibitors have also been shown to kill EBV-infected B cells and T cells by reducing the levels of EBV EBNA1 and LMP1. In 1 study, treatment of cells with the HSP90 ganetespib also reduced the level of phosphorylated Akt, delayed the onset of EBV+ B-cell lymphoma, and prolonged survival in SCID mice.31 Treatment of a patient with T-cell CAEBV with ganetespib reduced the percentage of EBV+ T cells in the peripheral blood.32