Ponderings and speculations about purinergic signaling, in pursuit of a unified ME/CFS theory

Learner1

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On the other hand, rather than focusing on every infection and sensitivity and allergy, I've been thinking about the big picture of a disordered response to things, recently. lots of people get lyme and are alright (although 20% of people get PTLDS to varying degrees, actually a high percentage). Lots of people get viruses and are alright. What's the problem with my system that I respond in such a way?
Some of us have a genetic predisposition to a dysfunctional immune system. Others have just encountered too many stressors which overload our bodies' natural defenses.
 

dreampop

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On the other hand, rather than focusing on every infection and sensitivity and allergy, I've been thinking about the big picture of a disordered response to things, recently. lots of people get lyme and are alright (although 20% of people get PTLDS to varying degrees, actually a high percentage). Lots of people get viruses and are alright. What's the problem with my system that I respond in such a way?
I thought this out of frustration, but it never really held up to scrutiny. It comes from the false assumption that things are well "ordered" before one was sick and in healthy people. But the truth is almost every one is running around with tons of problems, gut flora imbalances, psychological stress, genetic mutations, allergies, exposure to virus and all that is interacting with their host defense/metablism/neurology - that's life (and no, it's not just modern life bs is hard/we wern't built for it). All this while indulging bad habits and lifestyle. That creates chronic fatigue sure, but not the devastation of what me/cfs is.
 
is there something upstream of purinergic signalling?

sincerely,
a total noob
ATP is the primary ligand for the receptor discussed here. Viral infection can cause dysregulation of ATP via intracellular calcium modulation.The usual suspects for CFS increase Ca2+ and ATP. These intracellular molecules are not modern drug targets.

From Viruses as Modulators of Mitochondrial Function:

"(HCMV) localizes to mitochondria [86] and causes the trafficking of Ca2+ from the ER to mitochondria at 4–6 hrs. After infection [87]. Active Ca2+ uptake by mitochondrion induces the production of ATP and other Ca2+ dependent enzymes accelerating virus replication, and a decrease in Ca2+ levels in the ER has antiapoptotic effects."
 
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NB - I'm a newb so I can't tag anyone in this post...or I just can't figure out how to do it!!

Firstly, necessary8 can I please acknowledge your considerable skill in writing this thread – must have burnt a shedload of energy you don’t have. Thank you so very much – your information clarified quite a few thought processes for me.

I’m a newbie here, but a longtime fan of Dr Nav and now fangirling over Dr Geoff Burnstock – father of purinergic signaling. So much so, that I have started a bach of biomedical science. But be gentle, I’m still trying to get my head around this shit.

I’ve been focusing on the P2Y2 receptor and associated studies. I’m starting to understand the edges of this science but the P2Y2 receptor fits my family’s symptomology (me: CFS, son: ASD) more than the P2X7 ATP proinflammatory model. As you so beautifully pointed out necessary8, the differences in the continuum of ME, fibro, CFS lays in which receptor is your poison, or as per Dr Nav’s latest paper on autism – which direction your M1 or M2 mito continuum points – and all its GTP-ases.

I like the idea that there is too much ATP on the pericellular halo. If CD39 is unable to hydrolyse a huge amount of ATP because the ecto-apyrases are overwhelmed, what happens? Yep, there might be a downregulated error of the CD39 enzyme or associated ATP-ases or an antibody - so many different theories as to why ATP cellular respiration works to a certain point, and then doesn’t. Which brings me to my point – apyrase. Why not use apyrase therapeutically /for research purposes to test the ATP/CDR theory rather than suramin – so many issues with this drug that seriously, is it worth it? And judging by the experience or other postees who have given suramin a go, it may well not be stopping the ATP onslaught and resultant PEM.

This study tweaked my interest. The researchers used a purine/ATP approach to sepsis and discussed suramin as a weak alternative to apyrase. https://www.nature.com/articles/cddis201470

In relation to antibodies crossing the BBB necessarily8 – with the identification of a lymph system within the brain, the previous finding that antibodies can not traverse the BBB is up in the air -its is not the only mode of entry to the brain - scarily all bets are off in the CNS. Your CD39 antibody theory is still well and truly alive. https://www.ncbi.nlm.nih.gov/pubmed/26030524
 
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Why not use apyrase therapeutically /for research purposes to test the ATP/CDR theory rather than suramin – so many issues with this drug that seriously, is it worth it? And judging by the experience or other postees who have given suramin a go, it may well not be stopping the ATP onslaught and resultant PEM.
I proposed this idea in Part 2 of my ponderings. See right here:
I’m also starting to wonder if it would be possible (and safe), to inject ME/CFS patients intramuscularly with recombinant CD39 itself. Or maybe with a different ecto-nucleotidase, which wouldn’t be blocked with the antibodies. It wouldn’t cure anyone, but if it caused some symptomatic relief that would prove eATP really is accumulating in those muscles. There’s a lot to consider here in terms of safety though, and I don’t feel qualified to make judgements on this.
Apyrase is just a non-specific name for a group of ecto-NTPDases.

There are some safety concerns with this approach. The paper you linked tested on mice, not on humans. I dont think recombinant ecto-NTPDases were ever tested on humans (if you have a study that did this, link it please). In this context, suramin is a much safer approach.

Another thing to consider is that, well, suramin should have worked too. The fact that it didnt, puts this whole theory into question.

In relation to antibodies crossing the BBB necessarily8 – with the identification of a lymph system within the brain, the previous finding that antibodies can not traverse the BBB is up in the air -its is not the only mode of entry to the brain - scarily all bets are off in the CNS. Your CD39 antibody theory is still well and truly alive. https://www.ncbi.nlm.nih.gov/pubmed/26030524
This is interesting, but the thing is - if antibodies could cross over to the brain, a lot of pathogens could do it to. So viruses and bacteries would be much more dangerous, would cause mental and neurodegenerative diseases. Also, if antibodies could cross to the brain normally, we would have detected that. There are many studies analyzing the proteome of cerebrospinal fluid. If there were antibodies there, we would have known a long time ago. So the new brain lymphatic system also inevitabily has to have its own from of BBB, and cannot be a clever way around to get into the brain.

To tag someone just type @ before their name.

Also, since I'm already writing a post, @debored13 can you please not quote my whole, long-ass multi-page post every time you want to say something about it? Or quote your own long post, when you want to add something to it? It kinda litters the thread. Just dont make long quotes. Thank you.
 
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Apyrase is patented (a non-patented version would be eliminated too quickly from circulation I assume). It does not have first-in-human testing done yet, Astra Zeneca has the rights and they have not started recruiting for phase I for stroke. AZ will refuse an IND for CFS even if applied for and properly funded by the NIH (they won't, an organization maybe could), even assuming it passes a Phase 1 (most optimistically July 2019) because stroke has a higher market share. Anti-CD39 is probably even even further down the development pipeline.
 

debored13

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Also, since I'm already writing a post, @debored13 can you please not quote my whole, long-ass multi-page post every time you want to say something about it? Or quote your own long post, when you want to add something to it? It kinda litters the thread. Just dont make long quotes. Thank you.
Sorry about that, was a little out of it and having a hard time figuring out how to quote specific sections
 

debored13

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So I was looking at what people use to bring those worms out of dauer state...

found this post on researchgate: https://www.researchgate.net/post/C...e_dauer_exit-dafachronic_analogues_substitues

some researcher was looking for dafachronic acid analogues to use to bring c. elegans worms out of dauer.

I found this paper, related: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4749721/


I wonder if there are chemicals that are not anti-purinergics that could possibly end this state, especially if, like Naviaux says, it's analogous to dauer.
Someone made another thread on NAD+ being used to bring worms out of dauer, and people have had success with it.
I need to reread Naviaux's two relevant papers, but it seems that these systems--purinergic signalling, metabolism, immune system... are complex enough that there might be more than one way to affect them.

Have there been any updates from Ron/OMF re: if they are making progress on finding out what the unknown molecule in the blood is?
 

debored13

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So I was looking at what people use to bring those worms out of dauer state...

found this post on researchgate: https://www.researchgate.net/post/C...e_dauer_exit-dafachronic_analogues_substitues

some researcher was looking for dafachronic acid analogues to use to bring c. elegans worms out of dauer.

I found this paper, related: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4749721/


I wonder if there are chemicals that are not anti-purinergics that could possibly end this state, especially if, like Naviaux says, it's analogous to dauer.
Someone made another thread on NAD+ being used to bring worms out of dauer, and people have had success with it.
I need to reread Naviaux's two relevant papers, but it seems that these systems--purinergic signalling, metabolism, immune system... are complex enough that there might be more than one way to affect them.

Have there been any updates from Ron/OMF re: if they are making progress on finding out what the unknown molecule in the blood is?
I don't know to what extent c. elegans is a directly useful model for humans, but surely since naviaux considers dauer a useful model for development, perhaps some of the chemicals which help bring them out of dauer are useful

this dafachronic acid is some kind of steroid that seems specific to being used in c. elegans, though. I wonder if there are analogous steroids in humans that would work--pregnenolone, maybe?
 

debored13

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Another thing to consider is that, well, suramin should have worked too. The fact that it didnt, puts this whole theory into question.
Is this necessarily true? The fact that a few people tried it (which btw, props for the courage), at what could be too-low doses, and didn't get relief, that doesn't seem like enough evidence for dismissing it. Also it seems like even if the idea of targeting purinergic receptors directly doesn't work, that isn't all the way back to square one. Naviaux's metabolomics study on CFS would still offer clues, perhaps there are other substances that can bring people out of the hypometabolic state, if it really is analogous to dauer.
 

debored13

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So, the only abnormalities in my labs besides viral titers my doc interpreted as high, were low immunoglobulin, high IgE, and high eosinophils. My doctor and naturopath both suggested that the eosinophils and IgE were connected, she suggested it's related to some mast cell thing.

Why do I bring this up? Well, I just started to explore the connections between histamine, mast cells, and purinergic signalling.

I found two papers so far that illuminate these connections. Also, apparently there's a journal just dedicated to purinergic signalling.

I don't really understand the connections so far, but I'm really just trying to gather information that may be relevant for a sharper mind to pick through. Will run it by my grandfather that did cancer research.

Also, as i mentioned earlier, the role of microglia and TLR4 in neuroinflammation that low dose naltrexone is thought to affect may be related to mast cells, I'm finding a lot of key words that are similar.

the abstract on the second paper:
"Mast cells are widely recognized as effector cells of allergic inflammatory reactions. They contribute to the pathogenesis of different chronic inflammatory diseases, wound healing, fibrosis, thrombosis/fibrinolysis, and antitumor immune responses. In this paper, we summarized the role of P2X and P2Y receptors in mast cell activation and effector functions. Mast cells are an abundant source of ATP which is stored in their granules and secreted upon activation. We discuss the contribution of mast cells to the extracellular ATP release and to the maintenance of extracellular nucleotides pool. Recent publications highlight the importance of purinergic signaling for the pathogenesis of chronic airway inflammation. Therefore, the role of ATP and P2 receptors in allergic inflammation with focus on mast cells was analyzed. Finally, ATP functions as mast cell autocrine/paracrine factor and as messenger in intercellular communication between mast cells, nerves, and glia in the central nervous system."
re: the bolded part, is it possible that some asthma drugs could be useful in CFS?
 

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debored13

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So, the only abnormalities in my labs besides viral titers my doc interpreted as high, were low immunoglobulin, high IgE, and high eosinophils. My doctor and naturopath both suggested that the eosinophils and IgE were connected, she suggested it's related to some mast cell thing.

Why do I bring this up? Well, I just started to explore the connections between histamine, mast cells, and purinergic signalling.



the abstract on the second paper:
"Mast cells are widely recognized as effector cells of allergic inflammatory reactions. They contribute to the pathogenesis of different chronic inflammatory diseases, wound healing, fibrosis, thrombosis/fibrinolysis, and antitumor immune responses. In this paper, we summarized the role of P2X and P2Y receptors in mast cell activation and effector functions. Mast cells are an abundant source of ATP which is stored in their granules and secreted upon activation. We discuss the contribution of mast cells to the extracellular ATP release and to the maintenance of extracellular nucleotides pool. Recent publications highlight the importance of purinergic signaling for the pathogenesis of chronic airway inflammation. Therefore, the role of ATP and P2 receptors in allergic inflammation with focus on mast cells was analyzed. Finally, ATP functions as mast cell autocrine/paracrine factor and as messenger in intercellular communication between mast cells, nerves, and glia in the central nervous system."
re: the bolded part, is it possible that some asthma drugs could be useful in CFS?
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2847274/

this article explores the relationship between IgE, mast cells, and eosinophils, which also has a relationship to gut issues. @necessary8, You said in your original post that the gut was still an area to fit into the theory. I know this is almost complicating the theory, but eosinophils have a lot to do with gut issues, and mast cells/IgE have something to do with purinergic signalling.
 

debored13

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So I was looking at what people use to bring those worms out of dauer state...

found this post on researchgate: https://www.researchgate.net/post/C...e_dauer_exit-dafachronic_analogues_substitues

some researcher was looking for dafachronic acid analogues to use to bring c. elegans worms out of dauer.

I found this paper, related: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4749721/


I wonder if there are chemicals that are not anti-purinergics that could possibly end this state, especially if, like Naviaux says, it's analogous to dauer.
Someone made another thread on NAD+ being used to bring worms out of dauer, and people have had success with it.
I need to reread Naviaux's two relevant papers, but it seems that these systems--purinergic signalling, metabolism, immune system... are complex enough that there might be more than one way to affect them.

Have there been any updates from Ron/OMF re: if they are making progress on finding out what the unknown molecule in the blood is?
http://www.pnas.org/content/104/12/5014

This is an article on dafachronic acid. It modulates c. elegans lifespan through nuclear receptor signalling. I didn't know much about nuclear receptors, so I looked into nuclear receptors in humans. The drugs that act on them seem to be hormones like estradiol or testosterone, or glucocorticoids. I know people have probably tried most of these things, but have we considered the possibility of a simple hormone, like progesterone or pregnenolone, prompting dauer exit?
 
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Another thing to consider is that, well, suramin should have worked too. The fact that it didnt, puts this whole theory into question.
I'm just now learning that a few people have experimented with Suramin on their own. Could someone direct me to where this was mentioned?
 
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Here's a new review paper summarising links from Adenosine to autoimmunity.

The Role of Extracellular Adenosine Generation in the Development of Autoimmune Diseases
F. Morandi,
1 A. L. Horenstein, 2 , 3 R. Rizzo, 4 and F. Malavasi 2 , 3
Author information ► Article notes ► Copyright and License information ► Disclaimer
Go to:
Abstract
Adenosine (ADO) is an immunosuppressive molecule, which suppresses the immune responses by interacting with specific receptors expressed by immune effector cells. ADO is produced from ATP through the enzymatic activities of CD39 and CD73. Alternatively, ADO can be generated starting from NAD+, which is metabolized by the concerted action of CD38, CD203a/PC-1, and CD73. The role of ADO in immunity has been characterized in the last years in
physiology and in pathological settings. This review examines a panel of reports focused on the functions of ADO in the context of human autoimmune/inflammatory diseases and the selected animal models. The final aim is to consider the role of adenosinergic ectoenzymes and ADO receptors as novel therapeutic targets for selected diseases.

For reference I post again this chart, showing reduced Adenosine in MECFS patients (data is from Naviaux)



Frm the conclusion of the paper

In the last years, several authors reported that extracellular ADO, produced by ectoenzymes belonging to the “classical” adenosinergic pathway (CD39 and CD73), might be involved in the control of the immune response in the context of human autoimmune/inflammatory diseases. Indeed, alterations of the expression and function of both CD39 and CD73 have been reported to be related to the onset of these diseases. On the contrary, overexpression of these molecules, as well as agonists of ADO receptors, is able to mitigate the immune responses and the inflammatory reaction that are crucial for the development of the disease.
 
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In the last years, several authors reported that extracellular ADO, produced by ectoenzymes belonging to the “classical” adenosinergic pathway (CD39 and CD73), might be involved in the control of the immune response in the context of human autoimmune/inflammatory diseases. Indeed, alterations of the expression and function of both CD39 and CD73 have been reported to be related to the onset of these diseases. On the contrary, overexpression of these molecules, as well as agonists of ADO receptors, is able to mitigate the immune responses and the inflammatory reaction that are crucial for the development of the disease.
@Murph, I keep coming back to phosphorylation and low adenosine - so I'm back on the coffee :) I've been reading the old original potato apyrase experiments which talk about ATP and the way it competitively interacts with ADP, AMP and adenosine. I wondered if this is the metabolic trap Phair was talking about - ATP getting in the way of other metabolic processes which require phosphorylation (such as ubiquinone), particularly at the PANX 1 channel. So as a little experiment, I started using glycyrrhetinic acid (licorice extract) as a PANX1 inhibitor - to slow down ATP into the extracellular space - and hence the p2x7 responsiveness. It's been a week (and I've started slow) I have less lactic acid and much more energy. I had a flare of what I think of as p2x3 responses - wide spread pain, trigeminal nerve issues, gut distention issues ect ect. I'm taking ALA for that but it takes awhile to take effect. All over, I'm feeling much better.
 
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@Murph, I keep coming back to phosphorylation and low adenosine - so I'm back on the coffee :) I've been reading the old original potato apyrase experiments which talk about ATP and the way it competitively interacts with ADP, AMP and adenosine. I wondered if this is the metabolic trap Phair was talking about - ATP getting in the way of other metabolic processes which require phosphorylation (such as ubiquinone), particularly at the PANX 1 channel. So as a little experiment, I started using glycyrrhetinic acid (licorice extract) as a PANX1 inhibitor - to slow down ATP into the extracellular space - and hence the p2x7 responsiveness. It's been a week (and I've started slow) I have less lactic acid and much more energy. I had a flare of what I think of as p2x3 responses - wide spread pain, trigeminal nerve issues, gut distention issues ect ect. I'm taking ALA for that but it takes awhile to take effect. All over, I'm feeling much better.
That level of biological understanding is above my head but I'm pleased it's working! Keep us posted.