Hi everybody,
After listening to Dr Prusty's recent interview, and after doing a bit of research, I really wonder why we don't focus more on the anti-purinergic therapies to correct these abnormal pathways.
I already compiled detailed data explaining to Dr. Phair on this thread, how when autistic murine models took suramin, all metabolites abnormalities that would be altered according to his itaconate hypothesis, did normalize, including Kreb's cycle intermediaries, oxidative stress markers such as GSH/GSSG and NADH/NAD+, and at least part of the innate response including the complement protein C1q. In this vein, someone posted on the same thread how suramin has demonstrated to inhibit the synthesis of Interferons, at least of INFbeta:
https://pubmed.ncbi.nlm.nih.gov/29558821/
Regarding Dr Prusty's theory, he explains how herpes virus reactivate and affect plasmatic B1 cells resident in the hematopoietic organs (mainly in the bone marrow), making them to secrete less natural immunoglobulins that would lead to less cleaning of celular debris, what would in turn lead to auto-immunity. An example of this would be low IgM against fibronectin1, high serum fibronectin1 and how all this would combine to cause both mitochondrial fission and hyperfussion (both part of the CDR process as explained by Dr. Naviaux).
So, what's on top of this cascade, viral reactivation or the perpetuated cell danger response caused by extracellular purines?
Well, it seems that when purinergic receptors are inhibited, many infections cannot reactivate/replicate/infect:
For example, when renal human cells are infected in vitro with HHV-6A virus, the infection is halted when a P2X7 antagonist is applied. Here, the intracellular calcium doesn't increase when the anti-purinergic drug is added, probably indicating that the CDR is effectively prevented within the cell:
https://www.frontiersin.org/articles/10.3389/fphar.2020.00096/full
Similarly, when fibroblasts are treated with the anti-purinergic kaempferol, the infection with CMV replication is impaired:
https://www.pnas.org/doi/10.1073/pnas.1907562116
In the same vein, here it is shown how the infection of astrocytes (glial cells) with HHV1 is an ATP-dependent process, necesary for neurons to get infected. Well, suramin shows to effectively inhibit HHV1 replication in astrocytes and neuronal infection. Interestingly, when ATP was inoculated, mimicking the infected astrocytes (which secret ATP to allow neuronal infection), neurons got infected as well. This clearly indicates that purinergic signal is necesary for infection and inflammation of neuroglia and neurons by HHV1:
https://onlinelibrary.wiley.com/doi/abs/10.1002/glia.23895
Many other pathogens seem to need the purinergic signal for the successful infection and replication process such as other viruses, including Zika virus, HIV or Sars-Cov-2, and also bacteria (Pneumococcus), parasites (toxoplasma) and even fungi:
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7378483/
The protein DRP-1 has also been shown to be key during HHV-6 reactivation, as it directly causes mitochondrial fragmentation. Well, suramin has been shown to inhibit Drp-1 expression as well:
https://jps.biomedcentral.com/articles/10.1007/s12576-019-00666-9
On the contrary, Dr. Prusty explained how the protein mitofusin-1, which maintains the mitochondria fused, decreased when gamma-globulins from ME/CFS patients were inoculated to healthy cells. Well, suramin also showed to increased mitofusin-1 levels in murine hepatocytes treated with LPS:
https://jps.biomedcentral.com/articles/10.1007/s12576-019-00666-9
On the other hand, if Prusty's theory is correct and if purinergic signal was upstream in the patho-physiological process, then gammaglobulins levels should be restored with anti-purinergic therapy. This seems to be true in the adquired autistic murine model developed by Naviaux. In this experiment total immunoglobulins increased by 20%:
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3596371/
Furthermore, suramin has been shown to reduce renal fibronectin1 expression in rats:
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3464039/
Note that I don't expect to see easy and fast benefits with anti-purinergic therapies. Actually, I am learning that high levels of purinergic receptors inhibition can be counter-productive and even dangerous. For example, the second study done with suramim in autistic children, showed that doses of 20 mg/kg didn't work, and only doses of 10 mg/kg did work (autistic children saw over 50% of overall improvement after 3 months on suramin).
This would explain why some ME/CFS who have tried suramin at this dose didn't see any benefits.
This idea seems validated from the results of the following study, where rats were given probenecid (a pannexin-1 inhibitor, which is also an anti-purinergic agent) before and after causing them cerebral Ischaemic damage. We see that doses of 1 mg/kg were far more effective than doses of 10 mg/kg.
This is a very intriguing question, as the dose of the probenecid that blocks 50% of the pannexin-1 receptor in vitro is 150 mcrM, and the dose of 1 mg/kg is probably much lower than 7 mcrM according to pharmacokinetic studies on rats. So, perhaps we just need a temporary inhibition of the purinergic receptors to achieve a steady recovery.
Also, there are many purinergic receptors, and we have no idea which ones we need to block and to what degree in order to restore homeostasis.
I am going to try this theory myself. I am going to measure the IL-1 beta levels of my PBMCs after stimulation with LPS and ATP. In humans studies this has been shown to be an accurate way to measure the degree of activation of the inflammasome, which should correlate well with the degree of the purinergic signal.
I will be taking probenecid and brilliant blue FCF dye (a food dye) and then test my hypothesis (I am also considering other pannexin-1 inhibitors such as spironolactone or glycyrrhetinic acid).I will aim to inhibit the IL1-beta secretion about 20-30% only, and see what happens.
(Note that @Hip already calculated that perhaps the brilliant blue dye could reach cells in sufficient amounts to be anti-purinergic:
https://forums.phoenixrising.me/thr...i-purinergic-therapy.52427/page-4#post-936102).
I really think this is going to take time and be a hard process. Healing with ME/CFS always is. I will be also taking GcMAF as an immune stimulator and I hope it can help to eradicate some of the infected cells which are contributing to perpetuate the CDR.
I'll let you guys know how my experiment goes. I'd really like to know your opinions on the subject.
Sergio
After listening to Dr Prusty's recent interview, and after doing a bit of research, I really wonder why we don't focus more on the anti-purinergic therapies to correct these abnormal pathways.
I already compiled detailed data explaining to Dr. Phair on this thread, how when autistic murine models took suramin, all metabolites abnormalities that would be altered according to his itaconate hypothesis, did normalize, including Kreb's cycle intermediaries, oxidative stress markers such as GSH/GSSG and NADH/NAD+, and at least part of the innate response including the complement protein C1q. In this vein, someone posted on the same thread how suramin has demonstrated to inhibit the synthesis of Interferons, at least of INFbeta:
https://pubmed.ncbi.nlm.nih.gov/29558821/
Regarding Dr Prusty's theory, he explains how herpes virus reactivate and affect plasmatic B1 cells resident in the hematopoietic organs (mainly in the bone marrow), making them to secrete less natural immunoglobulins that would lead to less cleaning of celular debris, what would in turn lead to auto-immunity. An example of this would be low IgM against fibronectin1, high serum fibronectin1 and how all this would combine to cause both mitochondrial fission and hyperfussion (both part of the CDR process as explained by Dr. Naviaux).
So, what's on top of this cascade, viral reactivation or the perpetuated cell danger response caused by extracellular purines?
Well, it seems that when purinergic receptors are inhibited, many infections cannot reactivate/replicate/infect:
For example, when renal human cells are infected in vitro with HHV-6A virus, the infection is halted when a P2X7 antagonist is applied. Here, the intracellular calcium doesn't increase when the anti-purinergic drug is added, probably indicating that the CDR is effectively prevented within the cell:
https://www.frontiersin.org/articles/10.3389/fphar.2020.00096/full
Similarly, when fibroblasts are treated with the anti-purinergic kaempferol, the infection with CMV replication is impaired:
https://www.pnas.org/doi/10.1073/pnas.1907562116
In the same vein, here it is shown how the infection of astrocytes (glial cells) with HHV1 is an ATP-dependent process, necesary for neurons to get infected. Well, suramin shows to effectively inhibit HHV1 replication in astrocytes and neuronal infection. Interestingly, when ATP was inoculated, mimicking the infected astrocytes (which secret ATP to allow neuronal infection), neurons got infected as well. This clearly indicates that purinergic signal is necesary for infection and inflammation of neuroglia and neurons by HHV1:
https://onlinelibrary.wiley.com/doi/abs/10.1002/glia.23895
Many other pathogens seem to need the purinergic signal for the successful infection and replication process such as other viruses, including Zika virus, HIV or Sars-Cov-2, and also bacteria (Pneumococcus), parasites (toxoplasma) and even fungi:
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7378483/
The protein DRP-1 has also been shown to be key during HHV-6 reactivation, as it directly causes mitochondrial fragmentation. Well, suramin has been shown to inhibit Drp-1 expression as well:
https://jps.biomedcentral.com/articles/10.1007/s12576-019-00666-9
On the contrary, Dr. Prusty explained how the protein mitofusin-1, which maintains the mitochondria fused, decreased when gamma-globulins from ME/CFS patients were inoculated to healthy cells. Well, suramin also showed to increased mitofusin-1 levels in murine hepatocytes treated with LPS:
https://jps.biomedcentral.com/articles/10.1007/s12576-019-00666-9
On the other hand, if Prusty's theory is correct and if purinergic signal was upstream in the patho-physiological process, then gammaglobulins levels should be restored with anti-purinergic therapy. This seems to be true in the adquired autistic murine model developed by Naviaux. In this experiment total immunoglobulins increased by 20%:
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3596371/
Furthermore, suramin has been shown to reduce renal fibronectin1 expression in rats:
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3464039/
Note that I don't expect to see easy and fast benefits with anti-purinergic therapies. Actually, I am learning that high levels of purinergic receptors inhibition can be counter-productive and even dangerous. For example, the second study done with suramim in autistic children, showed that doses of 20 mg/kg didn't work, and only doses of 10 mg/kg did work (autistic children saw over 50% of overall improvement after 3 months on suramin).
This would explain why some ME/CFS who have tried suramin at this dose didn't see any benefits.
This idea seems validated from the results of the following study, where rats were given probenecid (a pannexin-1 inhibitor, which is also an anti-purinergic agent) before and after causing them cerebral Ischaemic damage. We see that doses of 1 mg/kg were far more effective than doses of 10 mg/kg.
This is a very intriguing question, as the dose of the probenecid that blocks 50% of the pannexin-1 receptor in vitro is 150 mcrM, and the dose of 1 mg/kg is probably much lower than 7 mcrM according to pharmacokinetic studies on rats. So, perhaps we just need a temporary inhibition of the purinergic receptors to achieve a steady recovery.
Also, there are many purinergic receptors, and we have no idea which ones we need to block and to what degree in order to restore homeostasis.
I am going to try this theory myself. I am going to measure the IL-1 beta levels of my PBMCs after stimulation with LPS and ATP. In humans studies this has been shown to be an accurate way to measure the degree of activation of the inflammasome, which should correlate well with the degree of the purinergic signal.
I will be taking probenecid and brilliant blue FCF dye (a food dye) and then test my hypothesis (I am also considering other pannexin-1 inhibitors such as spironolactone or glycyrrhetinic acid).I will aim to inhibit the IL1-beta secretion about 20-30% only, and see what happens.
(Note that @Hip already calculated that perhaps the brilliant blue dye could reach cells in sufficient amounts to be anti-purinergic:
https://forums.phoenixrising.me/thr...i-purinergic-therapy.52427/page-4#post-936102).
I really think this is going to take time and be a hard process. Healing with ME/CFS always is. I will be also taking GcMAF as an immune stimulator and I hope it can help to eradicate some of the infected cells which are contributing to perpetuate the CDR.
I'll let you guys know how my experiment goes. I'd really like to know your opinions on the subject.
Sergio
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