Unfolded Protein Response and A Possible Treatment for CFS

SlamDancin

Senior Member
Messages
570
@Murph
@datadragon

As Datadragon mentioned and I’m seeing in the literature is there is a known connection between IFNs and ER function/UPR and I found a 2020 review on the subject that’s pretty interesting;

Type I interferons and endoplasmic reticulum stress in health and disease

Type I interferons (IFNs) comprise of pro-inflammatory cytokines created, as well as sensed, by all nucleated cells with the main objective of blocking pathogens-driven infections. Owing to this broad range of influence, type I IFNs also exhibit critical functions in many sterile inflammatory diseases and immunopathologies, especially those associated with endoplasmic reticulum (ER) stress-driven signaling pathways. Indeed, over the years accumulating evidence has indicated that the presence of ER stress can influence the production, or sensing of, type I IFNs induced by perturbations like pattern recognition receptor (PRR) agonists, infections (bacterial, viral or parasitic) or autoimmunity. In this article we discuss the link between type I IFNs and ER stress in various diseased contexts. We describe how ER stress regulates type I IFNs production or sensing, or how type I IFNs may induce ER stress, in various circumstances like microbial infections, autoimmunity, diabetes, cancer and other ER stress-related contexts.
 

Violeta

Senior Member
Messages
3,191
@Murph
@datadragon

As Datadragon mentioned and I’m seeing in the literature is there is a known connection between IFNs and ER function/UPR and I found a 2020 review on the subject that’s pretty interesting;

Type I interferons and endoplasmic reticulum stress in health and disease

Type I interferons (IFNs) comprise of pro-inflammatory cytokines created, as well as sensed, by all nucleated cells with the main objective of blocking pathogens-driven infections. Owing to this broad range of influence, type I IFNs also exhibit critical functions in many sterile inflammatory diseases and immunopathologies, especially those associated with endoplasmic reticulum (ER) stress-driven signaling pathways. Indeed, over the years accumulating evidence has indicated that the presence of ER stress can influence the production, or sensing of, type I IFNs induced by perturbations like pattern recognition receptor (PRR) agonists, infections (bacterial, viral or parasitic) or autoimmunity. In this article we discuss the link between type I IFNs and ER stress in various diseased contexts. We describe how ER stress regulates type I IFNs production or sensing, or how type I IFNs may induce ER stress, in various circumstances like microbial infections, autoimmunity, diabetes, cancer and other ER stress-related contexts.
👀
 

SlamDancin

Senior Member
Messages
570
There’s definitely something to all this. Trying to tie it all together might be a little complicated but here’s another interesting possibility to connect.

A 2022 study in worms (unfortunately) did show that increasing Itaconate concentrations activated the mitochondrial UPR and actually caused increased lifespan

Itaconate prolongs the healthy lifespan by activating UPRmt in Caenorhabditis elegans

Could increased Itaconate plus a possible deficient UPR be what’s caused the bad steady state?
 

datadragon

Senior Member
Messages
424
Location
USA
Interleukin-6 (IL-6) up-regulates the ZIP14 gene expression ZIP14 (Slc39a14)., 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://www.pnas.org/doi/10.1073/pnas.0502257102

Cellular stress induced by the abnormal accumulation of unfolded or misfolded proteins at the endoplasmic reticulum (ER). Unresolved endoplasmic reticulum (ER) stress corresponds with various chronic diseases, including cancer, diabetes, obesity and neurodegeneration, hepatic steatosis. Although cellular zinc deficiency has been implicated in causing ER stress, the effect of disturbed zinc homeostasis on hepatic ER stress and a role for zinc during stress are unclear. This study reveals that ER stress increases hepatic zinc accumulation via enhanced expression of metal transporter ZIP14. Unfolded protein response-activated transcription factors ATF4 and ATF6α regulate Zip14 expression in hepatocytes. (During ER stress, the UPR-activated transcription factors ATF4 and ATF6α transcriptionally up-regulate Zip14 expression.) During ER stress, ZIP14-mediated zinc transport is critical for preventing prolonged apoptotic cell death and steatosis, thus leading to hepatic cellular adaptation to ER stress. These results highlight the importance of normal zinc transport for adaptation to ER stress and to reduce disease risk. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5530682/

Expanded quote from above:
Restriction of zinc from cells can induce ER stress, indicating that zinc is essential to maintain normal ER function. However, a role for zinc during hepatic ER stress is largely unknown despite important roles in metabolic disorders, including obesity and nonalcoholic liver disease. We have explored a role for the metal transporter ZIP14 during pharmacologically and high-fat diet–induced ER stress using Zip14−/− (KO) mice, which exhibit impaired hepatic zinc uptake. Here, we report that ZIP14- mediated hepatic zinc uptake is critical for adaptation to ER stress, preventing sustained apoptosis and steatosis. Impaired hepatic zinc uptake in Zip14 KO mice during ER stress coincides with greater expression of proapoptotic proteins. ER stress-induced Zip14 KO mice show greater levels of hepatic steatosis due to higher expression of genes involved in de novo fatty acid synthesis, which are suppressed in ER stress-induced WT mice. During ER stress, the UPR-activated transcription factors ATF4 and ATF6α transcriptionally up-regulate Zip14 expression. We propose ZIP14 mediates zinc transport into hepatocytes to inhibit protein-tyrosine phosphatase 1B (PTP1B) activity, which acts to suppress apoptosis and steatosis associated with hepatic ER stress. Zip14 KO mice showed greater hepatic PTP1B activity during ER stress. These results show the importance of zinc trafficking and functional ZIP14 transporter activity for adaptation to ER stress associated with chronic metabolic disorders

IFN-y is also induced by intense exercise, https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4849644/ and https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5484649/

IFN-y increases expression of GRP78, which is the same GRP78 that increases WASF3 levels. The 24 hr delay for PEM correlates with IFN-y levels that are known to rise ~24 hrs after exertion. https://forums.phoenixrising.me/thr...s-chronic-fatigue-syndrome.90582/post-2443910

As Datadragon mentioned and I’m seeing in the literature is there is a known connection between IFNs and ER function/UPR
Interferon-y (intensive exercise 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 uptake. NLRP3 activation also leads to IL-1b (and Il-6) and IL-18. Interleukin-18 (IL-18) synergizes with IL-2 to enhance cytotoxicity, interferon-gamma (IFN-y) production, and expansion of natural killer cells.

The protein WASF3 is boosted in response to ER Stress as one reason, and this disrupts the cells energy production when WASF3 was increased, while blocking the WASF3 increase through blocking ER Stress lowered WASF3 levels and restored mitochondrial function. They discovered that the ME/CFS patients had higher levels of WASF3, lower levels of Mitochondrial cytochrome c oxidase (MTCO1) (a sign that oxygen wasn’t being metabolized well) and higher levels of the ER stress protein, PERK. https://medicalnewsbulletin.com/jou...c-encephalomyelitis-chronic-fatigue-syndrome/ Note Cytochrome c oxidase, the terminal oxidase in the electron transport chain, is copper dependent which requires ceruloplasmin to make our copper and iron bio-available (usable). Zinc is involved in Vitamin A metabolism and vice versa and Zinc, Vitamin A ,and Magnesium are needed for ceruloplasmin production that Ive covered more elsewhere. Other mechanisms may also be at play.

Glucose deprivation strongly inhibited IFN-gamma gene expression (increased by ifn-a, heavy exercise), optimal induction of IFN-gamma transcription is a glucose-dependent process. IFN-γ rapidly increases protein synthesis and causes the unfolded protein response (UPR), as evidenced by the increased expression of glucose-regulated protein 78 (GRP78). IFN-y is also upregulated from intensive exercise.
https://forums.phoenixrising.me/thr...s-chronic-fatigue-syndrome.90582/post-2443910

High glucose treatment, but not the osmotic control mannitol, induces csGRP78 expression through an ER stress–dependent mechanism (which increases WASF3 levels). There are many 'roads' to increase NLRP3 and/or ER Stress. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6514638/ GRP78 levels were positively correlated with HbA1c and AGEs. https://pubmed.ncbi.nlm.nih.gov/34591271/
 
Last edited:

SlamDancin

Senior Member
Messages
570
They discovered that the ME/CFS patients had higher levels of WASF3, lower levels of Mitochondrial cytochrome c oxidase (MTCO1) (a sign that oxygen wasn’t being metabolized well) and higher levels of the ER stress protein, PERK.
@datadragon I can’t remember where I saw it now but the latest update from Dr Phair re: The Itaconate hypothesis, I believe, had some slides about oxygen metabolism defects. I’d love to go over the details if anyone knows
 

datadragon

Senior Member
Messages
424
Location
USA
@datadragon I can’t remember where I saw it now but the latest update from Dr Phair re: The Itaconate hypothesis, I believe, had some slides about oxygen metabolism defects. I’d love to go over the details if anyone knows
Hi, see below Zinc is needed for A20 aka TNF alpha-induced protein 3 TNFAIP3 that negatively regulates the itaconate shunt I posted in the WASF3 thread and tagged Dr Phair but never heard anything as its all linked together actually.

Zinc deficiency evokes the endoplasmic reticulum (ER)-stress response https://pubmed.ncbi.nlm.nih.gov/23748779/

SOD1 as a molecular switch for initiating the homeostatic ER stress response under zinc deficiency https://pubmed.ncbi.nlm.nih.gov/24076220/

Zinc is needed for A20 aka TNF alpha-induced protein 3 TNFAIP3 that negatively regulates the itaconate shunt. The zinc finger protein A20 is a tumor necrosis factor (TNF)- and interleukin 1 (IL-1)-inducible protein that negatively regulates nuclear factor-kappa B (NF-kappaB)-dependent gene expression. The mitochondrial enzyme aconitate decarboxylase 1 (ACOD1, best known as immunoresponsive gene 1 [IRG1]) is upregulated under various inflammatory conditions and serves as a pivotal regulator of immunometabolism involved in itaconate production, macrophage polarization, inflammasome activation, and oxidative stress. Under stress conditions, especially inflammatory stimulation, the expression of ACOD1 is upregulated by macrophages, monocytes, and DCs in the innate immunity system. The expression of ACOD1 is also upregulated in the tissue under infection, The upregulated ACOD1 also acts as a feedback mechanism to regulate the activation of transcription factors. For example, lipid A induces Acod1 mRNA upregulation by activating two transcription factors, namely nuclear factor kappa B subunit 1 (NFKB1) and interferon regulatory factor 3 (IRF3), while the increased expression of ACOD1 inhibits NFKB1 and IRF3, leading to LPS tolerance. This negative feedback mechanism between ACOD1 and NFKB1 can be mediated by the deubiquitinase TNF alpha-induced protein 3 (TNFAIP3, also known as A20), which inhibits NFKB1 activation and subsequent ACOD1 expression in myeloid cells in response to LPS, TNF, or carbon monoxide (CO). Increased ACOD1 expression limits NFKB1 activation by sustaining the expression of TNFAIP3. In addition, ACOD1-mediated itaconate production leads to the expression of activating transcription factor 3 (ATF3), thereby inhibiting the translation of the NFKB inhibitor zeta (NFKBIZ) and subsequent interleukin-6 (IL6) expression https://www.sciencedirect.com/science/article/pii/S2667100X22000147 https://pubmed.ncbi.nlm.nih.gov/10385526/

In vitro studies have shown that zinc decreases NF-κB activation and its target genes, such as TNF-α and IL-1β, and increases the gene expression of A20 and PPAR-α, the two zinc finger proteins with anti-inflammatory properties. Studies have demonstrated that physiological reconstitution of zinc restrains immune activation, whereas zinc deficiency, in the setting of severe infection, provokes a systemic increase in NF-κB activation. https://pubmed.ncbi.nlm.nih.gov/28083748/

Zinc inhibits the transactivation activity of NF-κB. Zinc inhibits iNOS promoter activity. https://www.sciencedirect.com/science/article/pii/S2213231714000834

Mechanistically, several immune receptors (e.g., TLRs and IFNAR), adapter proteins (e.g., MYD88), ubiquitin ligases (e.g., A20), and transcription factors (e.g., NF-κB, IRFs, and STATs) form complex signal transduction networks to control ACOD1 expression in a context-dependent manner. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7395145/

HIF1a is downstream of ACOD1, and HIF1a increases WASF3 also (see pattysmith post image on first page of wasf3 thread). So I just linked the itaconate shunt to increase in WASF3.

1-s2-0-s2667100x22000147-gr2-jpg.52563


Posted here: https://forums.phoenixrising.me/thr...s-chronic-fatigue-syndrome.90582/post-2443553
 
Last edited:

Violeta

Senior Member
Messages
3,191
In particular, WASF3 is a hypoxia-inducible gene, meaning that it contains at least one or more hypoxia responsive elements (HRE) which act as binding sites for transcription factors [23]. The levels of WASF3 protein increased significantly in MDA-MB-231 and MCF7 breast cancer cells incubated under hypoxic conditions.

The mitochondrial respiratory chain and cytochrome c oxidase (Cco) have been implicated in the induction of some hypoxic nuclear genes (hypoxic signaling) in both yeast and mammalian cells (13–16) exposed to reduced oxygen levels, and it has been proposed that mitochondrially generated ROS are involved (17–20)

From a by member lassesen
https://forums.phoenixrising.me/blog-articles/treatments-odds-of-remission-for-cfs-me.2434/
  • “In November, 1999, Dr. Joe Brewer (an Infectious Disease specialist in Kansas City) and I developed a model of pathogen activation of the immune and coagulation systems. The model proposes that the end result of such pathogenmediated activation is increased blood viscosity due to 1) an underlying coagulation regulatory protein defect, and 2) activation of the coagulation system by the pathogen. As the blood viscosity increases, the diminished blood flow creates hypoxia (lack of oxygen) and nutrient deprivation within various areas of the body. This is like trying to start your car in Wisconsin in the winter with 60- weight engine oil. This model explains the multi-organ symptomatology and also explains why the low dose heparin therapy is effective by increasing blood flow as the blood viscosity decreases. Thus, patients gain relief from their symptoms with this therapy.” [2000 Transcript]
  • Improvement: 90% or better
Any thoughts on hypoxia?

I am thinking blood viscosity.
 
Last edited:

Murph

:)
Messages
1,803
What I'm interested in most when it comes to the UPR is not necessarily the onset of it - that's well understood. The UPR is turned on when unfolded proteins accumulate. What's really captured my imagination is the second part. The part where the UPR decides thing have gone on long enough and starts the murder.

It kills cells via apoptosis (orderly cell death, lots of neat recycling) but also necrosis (kaboom, cell bits go everywhere). The Wust paper from the other day reported a lot of necrosis in muscle tissue after exercise and I have copied below the abstract from a paper that shows the UPR (or a version of it at least) can cause necrotic cell death

What's exciting about the second part, where cells start dying, is it happens after a delay. I'm REALLY struggling to find out how long that delay is, but I'm starting to get suspicious that it might be equivalent to the delay we see for PEM.

So my model of PEM is: exercise creates a backlog of unfolded proteins. The body turns on the appropriate response. That doesn't work. it moves to a new bad phase where it kills a lot of cells. We feel terrible. The appropriate interventions would be ones that increase ER capacity or blunt the transition to apoptosis/necrosis.


Strong and sustained activation of the anticipatory unfolded protein response induces necrotic cell death


Mara Livezey 1 , Rui Huang 1 , Paul J Hergenrother , David J Shapiro 3
DOI: 10.1038/s41418-018-0143-2

Abstract


The endoplasmic reticulum stress sensor, the unfolded protein response (UPR), regulates intracellular protein homeostasis. While transient activation of the reactive UPR by unfolded protein is protective, prolonged and sustained activation of the reactive UPR triggers CHOP-mediated apoptosis.

In the recently characterized, evolutionarily conserved anticipatory UPR, mitogenic hormones and other effectors pre-activate the UPR; how strong and sustained activation of the anticipatory UPR induces cell death was unknown. To characterize this cell death pathway, we used BHPI, a small molecule that activates the anticipatory UPR through estrogen receptor α (ERα) and induces death of ERα+ cancer cells. We show that sustained activation of the anticipatory UPR by BHPI kills cells by inducing depletion of intracellular ATP, resulting in classical necrosis phenotypes, including plasma membrane disruption and leakage of intracellular contents.

Unlike reactive UPR activation, BHPI-induced hyperactivation of the anticipatory UPR does not induce apoptosis or sustained autophagy. BHPI does not induce CHOP protein or PARP cleavage, and two pan-caspase inhibitors, or Bcl2 overexpression, have no effect on BHPI-induced cell death. Moreover, BHPI does not increase expression of autophagy markers, or work through recently identified programmed-necrosis pathways, such as necroptosis. Opening of endoplasmic reticulum IP3R calcium channels stimulates cell swelling, cPLA2 activation, and arachidonic acid release. Notably, cPLA2 activation requires ATP depletion.

Importantly, blocking rapid cell swelling or production of arachidonic acid does not prevent necrotic cell death. Rapid cell death is upstream of PERK activation and protein synthesis inhibition, and results from strong and sustained activation of early steps in the anticipatory UPR. Supporting a central role for ATP depletion, reversing ATP depletion blocks rapid cell death, and the onset of necrotic cell death is correlated with ATP depletion. Necrotic cell death initiated by strong and sustained activation of the anticipatory UPR is a newly discovered role of the UPR.
 

Gondwanaland

Senior Member
Messages
5,100
In addition to Fib-4 there is another calculator:

https://www.hepatitisc.uw.edu/page/clinical-calculators/apri

AST to Platelet Ratio Index (APRI) Calculator
This is an AST to Platelet Ratio Index (APRI) calculator tool. Enter the required values to calculate the APRI value. The APRI Score will appear in the oval on the far right (highlighted in yellow). Most experts recommend using 40 IU/L as the value for the AST upper limit of normal when calculating an APRI value.


Interpretation:
In a meta-analysis of 40 studies, investigators concluded that an APRI score greater than 1.0 had a sensitivity of 76% and specificity of 72% for predicting cirrhosis. In addition, they concluded that an APRI score greater than 0.7 had a sensitivity of 77% and specificity of 72% for predicting significant hepatic fibrosis.1
For detection of cirrhosis, using an APRI cutoff score of 2.0 was more specific (91%) but less sensitive (46%). The lower the APRI score (less than 0.5), the greater the negative predictive value (and ability to rule out cirrhosis) and the higher the value (greater than 1.5) the greater the positive predictive value (and ability to rule in cirrhosis); midrange values are less helpful. The APRI alone is likely not sufficiently sensitive to rule out significant disease. Some evidence suggests that the use of multiple indices in combination (such as APRI plus FibroTest) or an algorithmic approach may result in higher diagnostic accuracy than using APRI alone.2
 
Back