Research Re Exosomes and ME/CSF

[Joy&K0$]

Here are some links that might help:

Cytosorbents - cytokine adsorber "Cytosorb"
- Presentation showing how cytokine adsorption works. This is from the Feb 2019 presentation. I believe this could be a viable short-term treatment for ME patients. See especially slides 7-14: https://cytosorbents.com/wp-content...bents-Investor-Presentation-February-2019.pdf
- Summary of the Cytosorb (cytokine adsorber) therapy: https://cytosorb-therapy.com/the-therapy/
- Cytosorb literature database: https://literature.cytosorb-therapy.com/

Aethlon Medical - Exosome adsorber
- This is the link I was referring to in the above post. https://seekingalpha.com/filing/4061856 Ignore the investor stuff, and jump down to, "The Mechanism of the Hemopurifier". You can also do a search (Control “F”) in this document for “exosomes” or "Hemopurifier".
- Here's a link to what Time Magazine had to say in 2014 about Aethlon and the Hemopurifier http://tinyurl.com/y4hdz4m3
- And here is a white paper that has interesting sections on how the Hemopurifier removes exosomes from the blood. Focused on cancer exosomes, but if you dig around their site, you'll see they also address exosomes that may be relevant to inflammatory conditions, Parkinsons, etc. http://tinyurl.com/yd33efjo

I think one or both of these could help us, if only to "flip the switch" immunologically.

 

[Wally]

[*]I have not been able to read all the posts in this thread, but I thought this paper published in 2018 might be interesting to add to the discussion re exosomes. This paper also caught my attention because of the reference to retroviruses, which is always an interesting topic when discussing ME/CSF research.
See, https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6187740/

“Exosome Biogenesis and Biological Function in Response to Viral Infections”

“Introduction. Exosomes are extracellular vesicles that originate as intraluminal vesicles during the process of multivescular body formation. Exosomes mediate intercellular transfer of functional proteins, lipids, and RNAs. The investigation into the formation and role of exosomes in viral infections is still being elucidated. Exosomes and several viruses share similar structural and molecular characteristics.. . .
Conclusion. Given the ever-growing roles and importance of exosomes in both host and pathogen response, this review will address the impact role of exosome biogenesis and composition after DNA, RNA virus, on Retrovirus infections. This review also will also address how exosomes can be used as therapeutic agents as well as a vaccine vehicles.”

[*Edited this post to make it easier to follow after other posts were merged into this thread. See, post/reply no.1 for info. re the merging of thread posts.]

 

[Wally]

[*]During the NIH 2 Day April 2019 Conference - “Accelerating Research on ME/CFS”, Cort Johnson sent out the following tweet.

Ron Davis - nanoneedle study accepted for publication , Idea that something in the plasma is whacking the ability of MECFS patients cells to produce energy = still holding strong; not a cytokine - looks like it may be an exosome.

https://twitter.com/i/web/status/1114227318845784064

It is unclear from this tweet if the reference to “exosomes”, as a possible suspect in the yet as unidentified particle in patients blood, relates to an educated guess made by Ron Davis, Cort Johnson or someone else. Since exosomes seem to be a hot topic that is popping up in research outside of ME/CFS, it would be pretty amazing if Ron’s new nanoneedle found ME/CFS patients achilles heel in an exosome.

[*Edited this post to make it easier to follow after other posts were merged into this thread. See, post/reply no.1 for info. re the merging of thread posts.]

 

[Snow Leopard]

Cue the speculation.

 

[Rufous McKinney]

I thought it would be an interesting one to start off this thread

I agree at the same time that 12 exosome Phd's are needed to figure out the simplest anything. So something may be up with exosomes. And perhaps they can be: filtered out. But obviously this is 2000 X 10 times more complicated than that. Its too bad we cannot get Phd's to spend one hour per day helping us figure things out.

I thought i would simply comment on the Cell Death thing. So several key ME theories include something up with cell death. Exosomes are likely involved in escorting material that needs to be disposed of, like dead cell remains, out of the body via the blood stream. So we could simply have MANY MANY more exosomes than, say someone with less of that going on. Or maybe our exosomes are very large. Taking up too much space, making the plasma like not flow well or be thicker or something.

So here I just found out about: Human Breast Milk Exosomes: are protective of the intestine and so here we see the Exosome is Good News. And Its just dawned on me, my daughter is giving birth around April 20: maybe I could actually: borrow some.

https://www.ncbi.nlm.nih.gov/pubmed/29991305

 

[Wally]

I am working on pulling together some other threads that have appeared in the past on the Forum that may add some interesting tidbits of information to ponder when viewed through the lens of this thread. My brain is moving very slowly these days, so it might take me awhile to complete this task. If this thread has caught your attention you might want to mark it to watch to see if there are any other jigsaw puzzle pieces that are unearthered to add to this discussion.

 

[junkcrap50]

From reading Wikipedia on Exosomes, it seems like miRNA is a big component of exosome cargo. Wasn't there a ME/CFS researcher studying miRNA?

Also, seems like exosomes can carry a lot of stuff. Also from wiki:

The protein content of a single exosome, given certain assumptions of protein size and configuration, and packing parameters, can be about 20,000 molecules.[12]

I'm worried of what's IN the exosome as that will likely be very difficult to pin down.

 

[junkcrap50]

On the topic of exosomes and potential short-term treatments, two technologies I’ve been eyeing that I believe hold tremendous promise – potentially as “proof of principle” to temporarily treat M.E. patients; and also maybe helpful to bring our most severe patients back from the brink. IMO these could be viable for an M.E. clinical trial, to do big data evaluation on patients before/after cytokine and/or exosome adsorption.

a) Cytosorb’s (CTSO) Cytosorb extracorporeal cytokine adsorber. American company that is going gangbusters in Germany especially, with the Cytosorb used in ICU’s to treat systemic sepsis. Still an early stage biotech, however with doubling year-over-year revenues. As a new poster, I can't post links yet, but will try to shortly. Or you can go to the Cytosorb website and check out their investor presentation and their Cytosorb user’s group (most active in Germany) that waxes poetic on the ability of this cartridge to filter out cytokines from the blood. Quite dramatic physiological responses in these critically ill ICU patients. If M.E. is so closely related to Systemic Inflammatory Response Syndrome and low-grade sepsis, maybe we're ideal candidates too?

b) Aethlon Medical’s Hemopurifier. Started off as an adsorber for viruses, now also targeting cancerous and non-malignant exosomes. You can google Seeking Alpha, and Aethlon for this filing. Will also try to post the link if my health allows later: "We are also investigating the ability of the Hemopurifier to capture glycosylated bacterial toxins and tumor-derived exosomes that promote cancer progression and treatment resistance. Additionally, we are the majority owner of Exosome Sciences, Inc. (ESI), a Company that is focused on the discovery of exosomal biomarkers to diagnose and monitor life-threatening disease conditions that may be current or future therapeutic targets for Aethlon Medical.

Do a search (Control “F”) in this document for “exosomes”, and take a look at all the patents they have on things like “methods for quantifying exosomes”; “methods for removal of exosomes”, etc.

FYI AEMD was one of Time Magazine’s feted ideas a couple of years ago. Lots of folks bought in, then the stock tanked again, so I'm posting from the clinical perspective, not with any investment advice . IMO both of these early-stage biotechs have some maturing to do, but their products are fascinating, and potentially relevant for the M.E. arena.

Both of these technologies (CTSO and AEMD) require a dialysis machine.

@Joy&K0$ you should write letters to both of these companies and explain to them how ME/CFS could be valuable targets for their technologies. Also tell them about Ron Davis's research and his new finding on exosomes.

 

[Rufous McKinney]

could be valuable targets for their technologies

And I think there are more of us than the statisical evidence suggests. Since, for instance, I am currently without an ME diagnosis altho I clearly meet the most rigorous diagnostics.....my insurance does not even consider me to have this illness. My chart says: SEID.

 

[junkcrap50]

https://forums.phoenixrising.me/thr...-an-exploratory-pilot-study.58966/post-973260
This thread is relevant here. From 1 year ago. Talks about Extracellular Vesicles as a diagnostic marker for CFS. Also, mentions Maureen Hanson at Cornell as an expert and research in vesicles, and has been looking at them for CFS. I'll quote a few highlight posts.

COMMENT: Wondering if these extracellular vesicles are that "something" that Ron Davis mentioned is different is the blood of people with SEID. Although the sample was small, the level of significance was pretty darn good.

https://www.tandfonline.com/doi/full/10.1080/20013078.2018.1453730

ABSTRACT
Chronic Fatigue Syndrome (CFS), also known as Myalgic Encephalomyelitis (ME) is an acquired, complex and multisystem condition of unknown etiology, no established diagnostic lab tests and no universally FDA-approved drugs for treatment. CFS/ME is characterised by unexplicable disabling fatigue and is often also associated with numerous core symptoms. A growing body of evidence suggests that extracellular vesicles (EVs) play a role in cell-to-cell communication, and are involved in both physiological and pathological processes. To date, no data on EV biology in CFS/ME are as yet available. The aim of this study was to isolate and characterise blood-derived EVs in CFS/ME. Blood samples were collected from 10 Spanish CFS/ME patients and 5 matched healthy controls (HCs), and EVs were isolated from the serum using a polymer-based method. Their protein cargo, size distribution and concentration were measured by Western blot and nanoparticle tracking analysis. Furthermore, EVs were detected using a lateral flow immunoassay based on biomarkers CD9 and CD63. We found that the amount of EV-enriched fraction was significantly higher in CFS/ME subjects than in HCs (p = 0.007) and that EVs were significantly smaller in CFS/ME patients (p = 0.014). Circulating EVs could be an emerging tool for biomedical research in CFS/ME. These findings provide preliminary evidence that blood-derived EVs may distinguish CFS/ME patients from HCs. This will allow offer new opportunities and also may open a new door to identifying novel potential biomarkers and therapeutic approaches for the condition.

From what I recall of Dr. Davis' research, they filtered the serum, removing components too big to pass through (microbes, etc), and I assume they used a second filter to pass the smaller components out. The 'something' was in the middle, which held 'protein sized stuff'.

The vesicles might contain proteins associated with ME/CFS, but they're held in comparatively big containers (the vesicles), so they should have been filtered out.

My guess is that these extracellular vesicles will turn out to be a result of ME/CFS, not a cause. Whatever is messing up our cells might be increasing the chance of vesicles forming and separating.

https://news.cornell.edu/stories/2017/09/94m-nih-grant-funds-chronic-fatigue-syndrome-center
The second project, led by Hanson, will compare the content of extracellular vesicles between individuals with ME/CFS and controls. Extracellular vesicles are membrane-surrounded structures that contain cargos of proteins, lipids, hormones and RNAs that can influence the functions of cells when they fuse with them. These vesicles are known to be released after exercise and could be involved in cell-to-cell signaling. Hanson and colleagues will examine the proteins, small molecules and RNA content of extracellular vesicles before and after exercise, to see if inflammatory signals from the vesicles could be contributing to disease symptoms.

http://neuroimmune.cornell.edu/research/vesicles-and-signaling/
(saying more or less the same thing as above)
By analyzing, in conjunction with physiological data, metabolites, circulating inflammatory molecules, and extracellular vesicle (EV) cargo in blood samples from before and after exercise sessions, we aim to uncover markers and mechanisms of post-exertional malaise in ME/CFS. Our broad survey of possible molecular responses to exercise will include inflammatory proteins and immunogenic mitochondrial DNA fragments, targeted and untargeted metabolomics of blood serum, and a detailed proteomic and metabolomic characterization of EVs. EVs are released into the circulation during exercise and could therefore contain biomarkers or contain cargo that plays an active role in mediating the abnormal response to physical activity in ME/CFS.

Long version of grant description:
http://grantome.com/grant/NIH/U54-NS105541-01-5841

This shows the size of the extracellular vesicles they found. The ones in MECFS patients are smaller. (nta = nanoparticle tracking analysis)

From what I read there are two main kinds of extracellular vesicles produced by cells that are not dying. The paper notes that because our ones are small and packed with protein that suggests they are Exosomses, not the other, larger kind, known as microvesicles.

ANother interesting thing I learned at that link above is that you can even detect extracellular vesicles in urine. So that could be a particularly low-impact way to collect a biomarker if these EVs do prove useful.

Pilot researchers note how "easy" it is to isolate circulating EVs.

 

[Jackb23]

“2014 study indicated that during EBV’s last gasp while undergoing lytic replication, the virus was pouring enough dUTPase into exosomes to produce major immune effects that supported or promoted the establishment/maintenance of further EBV infections.“

http://simmaronresearch.com/2018/11...ting-chronic-fatigue-syndrome-mecfs-diseases/




“Abstract

We have recently demonstrated that Epstein-Barr virus (EBV)-encoded deoxyuridine triphosphate nucleotidohydrolase (dUTPase) modulates innate immunity in human primary monocyte-derived macrophages through toll-like receptor (TLR) 2 leading to NF-κB activation and the production of pro-inflammatory cytokines. Our previous depletion studies indicated that dendritic cells (DCs) may also be a target of the EBV-encoded dUTPase. However, the role of EBV-encoded dUTPase in DC activation/function and its potential contribution to the inflammatory cellular milieu characteristic of EBV-associated diseases remains poorly understood. In the present study, we demonstrate that EBV-encoded dUTPase significantly altered the expression of genes involved in oncogenesis, inflammation and viral defense mechanisms in human primary DCs by microarray analysis. Proteome array studies revealed that EBV-encoded dUTPase modulates DC immune responses by inducing the secretion of pro-inflammatory TH1/TH17 cytokines. More importantly, we demonstrate that EBV-encoded dUTPase is secreted in exosomes from chemically induced Raji cells at sufficient levels to induce NF-κB activation and cytokine secretion in primary DCs and peripheral blood mononuclear cells (PBMCs). Interestingly, the production of pro-inflammatory cytokines in DCs and PBMCs was TLR2-dependent. Together these findings suggest that the EBV-encoded dUTPase may act as an intercellular signaling molecule capable of modulating the cellular microenvironment and thus, it may be important in the pathophysiology of EBV related diseases.”

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3718799/


I know that for many people, their me/cfs was triggered by mono. I know that for many others, this wasn’t the case, but found this interesting.

 

[Rufous McKinney]

what's IN the exosome

And whats ON exosomes....apparently markers can attach to them.

Isn't it simply amazing we are alive? That bodies do any of this? That it works out as often as it does?

 

[Wally]

[*Edit - I have now been able to review Ron’s talk at the NIH Conference. During his talk Ron stated that the “particle” might be an exosome, but he did not identify the specific measurement/size of the particle.]
____________________________________________________________________________________________________

I thought I read or heard Ron Davis provide information on the specific size (measurement) of the particle that he found in the plasma of patients. Does anyone know if this information has been released publicly and if it has what is the size of the mystery particle other than it is a “very large particle”?

If the particle Ron found using nanotechnology is “very large” and nano particles go up in size to 100nm, I would think a very large particle could be around 100nm in size.
See, https://www.britannica.com/science/nanoparticle re nano particle measurements.

[*Edit - I have now been able to review Ron’s talk at the NIH Conference. During his talk Ron stated that the “particle” might be an exosome.]
Has Ron postulated that he thinks the “mystery” particle could be an exosome? If yes, do you know when and to whom he made such a statement? I know in the interview that was recorded with Ben last week that Ron only referred to the mystery object as a “a very large particle”. See, video linked in this thread (starting at 16:14 minute marker) -https://forums.phoenixrising.me/thr...prof-ron-davis-april-2019.75837/#post-2196740

Also on the thread with Ron’s April video update there were posts asking if the mystery particle was an exosome. Someone responded with the words “spoiler alert and the abbreviation “LPS”, which I believe is the abbreviation for “lipopolysaccharides”. Not sure if this post was just a guess as to what the particle might be (i.e. an exosome from a LPS activated macrophage) or it was actual info. released by Ron or his research team. It is interesting that in a 2014 paper re the “functional significance of macrophage-derived exosomes in inflammation and pain”, the exosomes were from LPS activated macrophages and they measured 100nm. See, https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4106699/ at page 13.

[*Edit - I have now been able to review Ron’s talk at the NIH Conference. During his talk Ron stated that the “particle” might be an exosome, but he did not identify the specific measurement/size of the particle.]
I have not been able to view any of the talks from the recent NIH Conference, so I was wondering if Ron or someone else said something at the conference that led Cort Johnson to use the word “exosome” in his 4/5/2019 tweet? I was also interested in knowing if the specific measurement of the “ very large particle” was discussed at the Conference?

 

[junkcrap50]

@Wally that's the thing. How large was the original description of "something in the blood" before the exosome mention. I've been having some trouble understanding the context of these sizes. Because previously, I thought that Ron put the size context as something below/smaller than bacteria (>400nm) and viruses (20-400nm), which he or others concluded were protein size (4-6nm). No mention of endotoxins (25nm-300nm) But exosomes (30-100nm) are much larger than proteins right? Since they carry thousands of them inside of it? So, it would really help try to get a more accurate picture of what size this "something in the blood" is.

 

[ljimbo423]

@Wally that's the thing. How large was the original description of "something in the blood" before the exosome mention. I've been having some trouble understanding the context of these sizes. Because previously, I thought that Ron put the size context as something below/smaller than bacteria (>400nm) and viruses (20-400nm), which he or others concluded were protein size (4-6nm). No mention of endotoxins (25nm-300nm) But exosomes (30-100nm) are much larger than proteins right? Since they carry thousands of them inside of it? So, it would really help try to get a more accurate picture of what size this "something in the blood" is.

This is the size of the filter Ron Davis used to filter out most of what was causing cellular dysfunction. Although it's not in size (nm) it's in weight.

.Big = larger than 10,000 molecular weight. Proteins, protein groups or antibodies (including autoantibodies). Amino acids are smaller, around 300 molecular weight.

http://forums.phoenixrising.me/inde...ow-up-to-2-21-17-research-update.49749/page-6 Post #115

 

[Joy&K0$]

@junkcrap50 sorry for the delay in getting back to you, and I appreciate and agree with your comment. Am just not able to take the flag on this one, but my gut feel is that an overture to a biotech about a possible ME/CFS trial should be made by an entity like OMF, or an eminent researcher in the feel (or a stinking-rich patient who could fund one LOL). My sense is that for treatment trials to be seriously entertained by a biotech, it would have far more impact for the idea to come from, say - the Director of the Stanford Genome Technology Center , or someone else who can really speak eloquently on the field (and with the academic/medical credibility to get 5+ minutes of their time). Also, my gut tells me this idea first has to pass muster with our scientist insiders in the ME/CFS field...? In the case of CTSO, I think if our researchers were to contact the head of the Cytosorbent's User's Group, or just take a gander at the studies on removal of inflammatory cytokines in sepsis, that might be an intriguing start. Or contact the Senior Scientist or Director of Research at Aethlon Medical?

Feel free to pass my posts on exosomes, cytokines and treatment to anyone you guys think might be able to run with this. If they pass muster with those in-the-know, I think these might be ideas worth pursuing by those who have a little more clout and understanding of this disease.

Either way, I'll keep digging, and posting ideas when I'm up to it.

 

[Wally]

@junkcrap50 - In listening to Ron’s talk at the conference, it is my understanding that the paper re the nanoneedle research was accepted for publication at the end of last week. Ron seemed to think the paper would be published “very soon”. Not sure what “very soon” equates to for this particular publisher/journal, but perhaps the paper will include the actual size of the mystery particle. Otherwise, I guess we will have to wait for a tweet, a post, a video etc... for Ron or whoever he authorizes to publicly release information re this particle’s measurements.

Not a scientist myself, so it is not like I am waiting to conduct my own nano particle experiments in my state of the art bedroom lab.⚒ Just a curious and lonely patient that finds this discovery to be very tantalizing and who hungers to learn more.

Patience and this illness does not come easily to me. After 28 years of being run over by a fleet of “Mack” trucks. my “patience” auxiliary tanks are running pretty low. So, I guess I will go do what I seem to do best and pull the covers back over my head , while I dream that hidden deep inside my brain I have all the right puzzle pieces to help solve this illness. (Cue the laugh track )

 

[Rufous McKinney]

⚒ Just a curious and lonely patient that finds this discovery to be very tantalizing and who hungers to learn more.

We are in a desert, parched and thirsty. More than 40 days and nites we have wandered. We search for the bread crumbs which might lead us out of the dark forest (no offense to dark forests which I generally love).

Where did you find such great Emojis!!!???

 

[Mel9]

And I think there are more of us than the statisical evidence suggests. Since, for instance, I am currently without an ME diagnosis altho I clearly meet the most rigorous diagnostics.....my insurance does not even consider me to have this illness. My chart says: SEID.

But I thought SEID is the same thing as MECFS?

 

[junkcrap50]

Wow this is new - at least to me. Maybe others didn't know this. According to Cort's latest HealthRIsing article (https://www.healthrising.org/blog/2019/04/15/nih-chronic-fatigue-lipkin-davis-prusty-oh/), Ron Davis has been able to recreate ME/CFS cells from healthy cells & healthy serum by adding an exosome. This is why he thinks it might be an exosome. Also another ME/CFS research is also looking at exosomes, in addition to Dr. Hanson. Emphasis added below:

Something in the blood appears to be taking a two-by-four to our cells. Studies indicate that it’s not a metabolite – our metabolism is not producing some metabolic breakdown component that is disturbing our cells. Nor does it appear to be a cytokine, but in what may be a very lucky break, Davis was able to recreate the problem with an exosome. Davis wouldn’t commit to it being an exosome, but right now that’s what it looks like. That would be a great break and good news, given that both Maureen Hanson and Prusty are examining exosomes.