Enterovirus and treatments

[Hip]

Interesting idea. I'm still unclear on exactly how the whole RNAi cascade is supposed to work. One paper I was reading made it sound like the end result of RNase deployment in a cell is apoptosis. If that were the case this probably wouldn't be very safe.

I did not realize that dicer is central to the process of RNA interference. So as you say, it probably wouldn't be very safe to use dicer in a therapy, unless dicer was specifically targeted at non-cytolytically infected cells, perhaps using the DRACO-like targeting system mentioned above.

Is this true though? I don't believe the cells that Chia finds dsRNA in inside the stomach tissue are quiescent.

That's a point. The enteroviral infections found in Dr Chia's stomach biopsies were presumably within epithelial cells, which are rapidly dividing cells, not quiescent cells. And Chia said in his study that he found non-cytolytic viruses in these biopsies.

However, Prof Nora Chapman states that these non-cytolytic infections, which have part of their genome deleted, are mainly created in quiescent cells. See page 21 of these presentation slides from Nora Chapman.

And I read elsewhere that in a dividing cell, the production of these non-cytolytic viruses that have part of their genome deleted is tiny in comparison to the production of viruses with full and intact genomes. In other words, in a dividing cell, most of the viruses produced are the cytolytic type, rather than the non-cytolytic type.

Although since non-cytolytic viruses may be able to transmit to and infect the surrounding cells, might it be that the non-cytolytic virus is originally created in a quiescent cell in the stomach, and then later spread to the stomach epithelial cells? I looked up the basement membrane cells on which epithelial cells sit, and apparently basement membrane cells normally are quiescent.


Update: in fact Chia says in his paper that the stomach cells he finds infected with enteroviruses are parietal cells, which I believe are quiescent.

 

[halcyon]

I've been thinking about this a bit. I believe I understand why the immune system has trouble identifying and clearing out the chronically infected cells.

When an enterovirus infects a cell, it triggers the formation of autophagosome like vesicles which the viral replication complex (the positive and negative strand viral RNA, polymerase, and host factors) moves into. This might shield the dsRNA from the host cell's pattern-recognition receptors (PRRs). For example, in order to bind to TLR7 or TLR3 receptors and trigger a cell mediated immune response, ssRNA or dsRNA must be internalized from the cytoplasm into an endosome containing the TLRs. Additionally, the IPS-1 pattern-recognition receptors MDA-5 and RIG-I can detect dsRNA and ssRNA in the cytoplasm, but none of these pathways will do much good when the dsRNA is protected inside a vesicle. There should be ssRNA present in the cytoplasm of chronically infected cells for the IPS-1 system to detect, but I'll get to that in a moment.

The end result of triggering the pattern-recognition receptors in a cell is production and release of IFN-β. Some of the released IFN-β will bind back on the cell in an autocrine manner and will trigger the IFN-stimulated genes. This upregulates the cell mediated defenses such as the 2'5'-OAS/RNase and PKR pathways, which are the most interesting when it comes to dealing with the dsRNA present in the infected cell.

As an added layer of defense though, enteroviral protease 2A and 3C (and possibly others) have the ability to interfere with both the TLR and IPS-1 pattern recognition pathways as well as the IFN stimulation pathway. So even if the cell does manage to detect the viral RNA, the virus can effectively halt the cell mediated response.

Another interesting aspect of the autophagosome vesicles and the ability of the virus to modulate autophagy is that there is some evidence that enterovirus might use this structure to leave the cell in a non-lytic manner. This might even allow the virus to move about between cells unmolested by neutralizing antibodies in the blood. This might partially explain how these chronic enterovirus infections can persist for long periods of time. Apparently it is difficult to prove experimentally that enterovirus can use this non-lytic mechanism but the possibility puts an interesting spin on things.

In light of all of this, I think the best approach is multifaceted and needs to include compounds that interfere with viral translation (e.g. Amantadine) as well as viral protease activity (e.g. fisetin, rutin), and also something to stimulate interferon production to bypass the lack of recognition of the vesicle bound dsRNA protected inside the cells. This assumes that the RNase can penetrate the autophagosome vesicles. I'm a little unclear on exactly which form of interferon is needed. It seems that it should be IFN-α or IFN-β as they both seem to bind to the same receptor (IFNAR) that triggers the cell mediated pathway, but I've seen little to no mention of IFN-β in this context. Dr. Chia seems to have only used synthetic IFN-α and IFN-γ. I believe oxymatrine only stimulates IFN-γ but I may be wrong about that. I need to do more reading on the interferon angle of this approach.

 

[Thomas]

I've been thinking about this a bit. I believe I understand why the immune system has trouble identifying and clearing out the chronically infected cells.

When an enterovirus infects a cell, it triggers the formation of autophagosome like vesicles which the viral replication complex (the positive and negative strand viral RNA, polymerase, and host factors) moves into. This might shield the dsRNA from the host cell's pattern-recognition receptors (PRRs). For example, in order to bind to TLR7 or TLR3 receptors and trigger a cell mediated immune response, ssRNA or dsRNA must be internalized from the cytoplasm into an endosome containing the TLRs. Additionally, the IPS-1 pattern-recognition receptors MDA-5 and RIG-I can detect dsRNA and ssRNA in the cytoplasm, but none of these pathways will do much good when the dsRNA is protected inside a vesicle. There should be ssRNA present in the cytoplasm of chronically infected cells for the IPS-1 system to detect, but I'll get to that in a moment.

The end result of triggering the pattern-recognition receptors in a cell is production and release of IFN-β. Some of the released IFN-β will bind back on the cell in an autocrine manner and will trigger the IFN-stimulated genes. This upregulates the cell mediated hosts defenses such as the 2'5'-OAS/RNase and PKR pathways, which are the most interesting when it comes to dealing with the dsRNA present in the infected cell.

As an added layer of defense though, enteroviral protease 2A and 3C (and possibly others) have the ability to interfere with both the TLR and IPS-1 pattern recognition pathways as well as the IFN stimulation pathway. So even if the cell does manage to detect the viral RNA, the virus can effectively halt the cell mediated response.

Another interesting aspect of the autophagosome vesicles and the ability of the virus to modulate autophagy is that there is some evidence that enterovirus might use this structure to leave the cell in a non-lytic manner. This might even allow the virus to move about between cells unmolested by neutralizing antibodies in the blood. This might partially explain how these chronic enterovirus infections can persist for long periods of time. Apparently it is difficult to prove experimentally that enterovirus can use this non-lytic mechanism but the possibility puts an interesting spin on things.

In light of all of this, I think the best approach is multifaceted and needs to include compounds that interfere with viral translation (e.g. Amantadine) as well as viral protease activity (e.g. fisetin, rutin), and also something to stimulate interferon production to bypass the lack of recognition of the vesicle bound dsRNA protected inside the cells. This assumes that the RNase can penetrate the autophagosome vesicles. I'm a little unclear on exactly which form of interferon is needed. It seems that it should be IFN-α or IFN-β as they both seem to bind to the same receptor (IFNAR) that triggers the cell mediated pathway, but I've seen little to no mention of IFN-β in this context. Dr. Chia seems to have only used synthetic IFN-α and IFN-γ. I believe oxymatrine only stimulates IFN-γ but I may be wrong about that. I need to do more reading on the interferon angle of this approach.

Right.

 

[Hip]

When an enterovirus infects a cell, it triggers the formation of autophagosome like vesicles which the viral replication complex (the positive and negative strand viral RNA, polymerase, and host factors) moves into.

Very interesting, I was not aware of that. This study found that inhibiting autophagosome formation significantly reduced coxsackievirus B3 replication.

 

[Hip]

In light of all of this, I think the best approach is multifaceted and needs to include compounds that interfere with viral translation (e.g. Amantadine) as well as viral protease activity (e.g. fisetin, rutin), and also something to stimulate interferon production to bypass the lack of recognition of the vesicle bound dsRNA protected inside the cells.

This study indicates that the supplements fisetin and rutin will inhibit the 3C protease (aka 3C proteinase) of enterovirus 71. However, I don't think this necessarily means these supplements can inhibit the 3C protease found in coxsackievirus B and echovirus, the enteroviruses associated with ME/CFS.

Though this study found the easy-to-obtain nitric oxide donor drugs nitroglycerin (aka: glyceryl trinitrate) and isosorbide dinitrate specifically inhibit the 3C protease of coxsackievirus B3, and the 2A protease of coxsackievirus B3. The study also found that in mice with coxsackievirus B heart muscle infection (myocarditis), treatment with either of these two drugs led to significantly reduced signs of myocarditis.

@zzz actually obtained full remission from his ME/CFS after a single dose of isosorbide dinitrate (a 10 mg tablet), and this remission lasted for nearly 8 years, before he unfortunately experienced a relapse. @zzz said that he had dramatic improvement in his symptoms within 3 days of taking this single 10 mg tablet: that is to say, he went from bedbound to going back to work in just 3 days. Full remission of all his ME/CFS symptoms occurred after 3 weeks of taking this single isosorbide dinitrate dose.

Though because @zzz's bedbound-to-healthy ME/CFS remission occured so fast, over just 3 days, that may be a little too quick to be due to purely antiviral effects, and may instead be due to some other properties of isosorbide dinitrate. Although antiviral effects can sometimes manifest very quickly: Dr Chia notes that with oxymatrine, improvements in ME/CFS symptoms can be very fast, occurring over a period of a few days when oxymatrine induces a fever, after which the patient feels much better.



After his relapse, when @zzz then tried isosorbide dinitrate once again, hoping it would fix him like it did the first time, the second time around this drug unfortunately had no effect. Assuming a viral explanation, this might be because the first time round, @zzz's ME/CFS was due to an enterovirus infection; but his relapse into ME/CFS 8 years later may have been caused by another virus (@zzz postulates a varicella link to his relapse), against which isosorbide dinitrate has no antiviral effect.

 

[acer2000]

Interesting, isn't isosorbide dinitrate used as a vasodialotor?

 

[Hip]

Interesting, isn't isosorbide dinitrate used as a vasodialotor?

Indeed, but it is also a drug very similar to the nitroglycerin drug Dr Jay Goldstein used for his ME/CFS patients, and in a small subset of patients, nitroglycerin apparently brought rapid remission. Both nitroglycerin and isosorbide dinitrate are nitric oxide donors (that's how they achieve vasodilation). It's possible that vasodilation is a mechanism by which these drugs sometimes induce remission from ME/CFS. @zzz's account of his remission after taking isosorbide dinitrate can be found on this thread:

Nitroglycerin (Glyceryl Trinitrate) And Dr Jay A. Goldstein's Other Instant Remission ME/CFS Treatme

 

[acer2000]

I will read that thread. Curious though because naphazoline also is used and it has the opposite (vasoconstricting) effect. The effect on me/cfs must be independent of these effects?

 

[halcyon]

This study indicates that the supplements fisetin and rutin will inhibit the 3C protease (aka 3C proteinase) of enterovirus 71. However, I don't think this necessarily means these supplements can inhibit the 3C protease found in coxsackievirus B and echovirus, the enteroviruses associated with ME/CFS.

The full paper mentions that they also work against CVB4 and echovirus 6 as well as poliovirus, rhinovirus, and hepatitis A virus. That's a pretty decent handful of picornaviruses.

As always I'm skeptical that oral administration will have a large impact, but I ordered some of both just to try out.

 

[halcyon]

Though this study found the easy-to-obtain nitric oxide donor drugs nitroglycerin (aka: glyceryl trinitrate) and isosorbide dinitrate specifically inhibit the 3C protease of coxsackievirus B3, and the 2A protease of coxsackievirus B3. The study also found that in mice with coxsackievirus B heart muscle infection (myocarditis), treatment with either of these two drugs led to significantly reduced signs of myocarditis.

I wonder how well some of the other NO supplements would work in place of nitroglycerin, things like arginine, AAKG, citrulline, etc. I've avoided these things so far, worrying that the vasodilation would make my OI symptoms worse. I'm also worried that the extra urea would piss off my kidneys.

Have you tried any of these NO supplements Hip?

 

[Hip]

I wonder how well some of the other NO supplements would work in place of nitroglycerin, things like arginine, AAKG, citrulline, etc.

I used to take the supplement citrulline malate 3 grams twice daily for years, as I found this helped with reducing my severe generalized anxiety disorder symptoms (this was before I discovered better anti-anxiety meds like N-acetyl-glucosamine).

I also used arginine pyroglutamate 5 grams twice daily for extended periods, for the same anti-anxiety purposes. Arginine pyroglutamate crosses the blood-brain barrier more easily that other forms of arginine.

I can't say that I noticed any improvements in my ME/CFS while taking these; they just seemed to work on anxiety.

 

[halcyon]

I used to take the supplement citrulline malate 3 grams twice daily for years, as I found this helped with reducing my severe generalized anxiety disorder symptoms (this was before I discovered better anti-anxiety meds like N-acetyl-glucosamine).

I also used arginine pyroglutamate 5 grams twice daily for extended periods, for the same anti-anxiety purposes. Arginine pyroglutamate crosses the blood-brain barrier more easily that other forms of arginine.

I can't say that I noticed any improvements in my ME/CFS while taking these; they just seemed to work on anxiety.

Did you have any OI symptoms during the time you started using these? If so, I'm assuming these didn't make those symptoms worse if you continued to take them.

 

[Hip]

Did you have any OI symptoms during the time you started using these?

Yes I have POTS, but these supplements do not seem to affect it.

Although having said that, I have never really been able to isolate my POTS symptoms from my ME/CFS symptoms, so it would be hard to know whether something was having an affect on my POTS. I don't get any specific symptoms on standing other than the 30 point increase in heart rate which is diagnostic for POTS. It's possible that POTS is contributing to my overall fatigue and brain fog, but if so, what percentage of my fatigue and brain fog comes from POTS, and what percentage comes from ME/CFS, I cannot tell.

That's probably a question I should ask other POTS patients here: how do they distinguish between POTS and ME/CFS symptoms.

 

[halcyon]

Yes I have POTS, but these supplements do not seem to affect it.

Interesting. Seems like it's worth a shot then, though I'm not sure which one would be the best to try.

Although having said that, I have never really been able to isolate my POTS symptoms from my ME/CFS symptoms, so it would be hard to know whether something was having an affect on my POTS. I don't get any specific symptoms on standing other than the 30 point increase in heart rate which is diagnostic for POTS. It's possible that POTS is contributing to my overall fatigue and brain fog, but if so, what percentage of my fatigue and brain fog comes from POTS, and what percentage comes from ME/CFS, I cannot tell.

I know exactly what you mean.

 

[Hip]

Arginine has been shown beneficial for coxsackievirus B myocarditis, so this might be a good bet.

You can also raise NO by taking nitrate supplements. I did some NO-raising experiments taking potassium nitrate 2 grams daily.

 

[knackers323]

Been any progress with enterovirus in last couple of years?

 

[JinZ]

Been any progress with enterovirus in last couple of years?

 

[Hip]

Been any progress with enterovirus in last couple of years?

Dr Chia recently completed a brain autopsy which showed enterovirus infection present throughout the brain on an ME/CFS patient. This is now the third brain autopsy which showed enterovirus infection in the brains of ME/CFS patients. See an article about these brain autopsies here.

Recently, probable enterovirus infection in the neurons of Parkinson's disease patients has been discovered, so that's another neurological disease that may be due to enterovirus.


You can see a list of diseases which have now been linked to chronic enterovirus infection in the Intro section of this article on non-cytolytic enterovirus (non-cytolytic enterovirus is the mutated form of enterovirus which causes chronic infections; normal unmutated enterovirus can only cause acute infections).


For a general discussion on why infectious pathogens will likely turn out to be the cause of most chronic diseases and cancer of currently unknown etiology, see some of the works of Prof Paul Ewald.

 

[perrier]

Dr Chia recently completed a brain autopsy which showed enterovirus infection present throughout the brain on an ME/CFS patient. This is now the third brain autopsy which showed enterovirus infection in the brains of ME/CFS patients. See an article about these brain autopsies here.

Recently, probable enterovirus infection in the neurons of Parkinson's disease patients has been discovered, so that's another neurological disease that may be due to enterovirus.


You can see a list of diseases which have now been linked to chronic enterovirus infection in the Intro section of this article on non-cytolytic enterovirus (non-cytolytic enterovirus is the mutated form of enterovirus which causes chronic infections; normal unmutated enterovirus can only cause acute infections).


For a general discussion on why infectious pathogens will likely turn out to be the cause of most chronic diseases and cancer of currently unknown etiology, see some of the works of Prof Paul Ewald.

Thank you Hip for posting this. If you have further information, what is Dr Chia suggesting be done? There are no meds which address enteroviruses, if I am understanding correctly. And is there any way of knowing if he has collaborated with the Stanford folks on his findings.

 

[Hip]

Thank you Hip for posting this. If you have further information, what is Dr Chia suggesting be done? There are no meds which address enteroviruses, if I am understanding correctly.

Oxymatrine, Epivir, tenofovir and interferon beta are the only medications currently used by Dr Chia to treat enterovirus ME/CFS (to my knowledge). These help some patients to some degree, but not others.

Interferon beta he only now uses for the severe bedbound hospitalized patients, and he finds that within just a few weeks of interferon, these patients are well enough to take long walks around the hospital.


But very good news is on the horizon: in around 2 years time, one or possibly two potent new anti-enterovirus drugs will become available. These new antiviral drugs, which are antiviral for coxsackievirus B, will likely make dramatic improvements and possibly cures in ME/CFS linked to CVB. Those drugs are already fully developed, and work well in animal studies; they just need to be put through human clinical trials before they are brought to market. Dr Chia says that they should be available to buy in around 2 years. More info here.