Ex-contractors for Moderna hand-wave that they know how to reduce the risk of mRNA and vehicle toxicity
But it is the stuff they CANNOT fix that wasn't addressed or partially addressed: A review of the Nature Paper
Recently, the following paper was published in Nature Reviews Drug Discovery by Dimitrious Bitounis et al. This researcher is a nanotoxicity researcher and also researches, ahem…..graphene oxide. GO alters gut microbe in vivo in humans
I will summarize parts of this review and highlight what I thought was important regarding risks of the mRNA-LNP therapy platform identified by the reviewers.
Translated mRNA therapies
Here the authors discuss how mRNA is immunogenic in itself and what can be done to decrease this immunogenicity. They discuss N-1-methylpseudouridine substitutions as the seminal event to decrease the immunogenicity of mRNA itself. They also discuss the immunogenicity of dsRNA on TLR signalling, and cytokine production. Unmodified in vitro mRNA produce pro-inflammatory cytokines including IL-12, interferon-alfa, TNF, IL-6 and others.
I was also pleased so see them spend some time on dsRNA and how to remove that from the mRNA drug substance. This contaminant is ofter overlooked due to the discovery of residual DNA contamination but it is critical to have it removed due to the issues above. Since it is Moderna, they are not using the patented method just found in a patent, as I discussed in my previous substack dsRNA, DNA and a BioNTech patent. Instead they mention using the cellulose method published by BioNTech, RP-HPLC for dsRNA removal and using the T7 polymerase computationally engineered to reduce the formation of dsRNA during the IVT process.
What was not mentioned?
endotoxin removal
residual dsDNA removal
nucleotide removal
other enzymes and buffers used in the IVT process
truncated and fragmented mRNA from the IVT process
codon optimization and mRNA secondary and tertiary structure
These are ongoing issues with mRNA purification from the IVT process. Instead, these researches suggest that CIRCULAR RNA was the answer to improving the relatively short half-life of linear mRNA
Circular RNA??
Circular mRNA (circRNA) is a single-standed mRNA molecule which was first discovered in plant viroids, and since then in mammalian cells. MOST recently, obelisks have been found which can translated a protein best explained in Kevin McKernans excellent substack on this issue. Viroids and Obelisks
Now, they are being promoted to be used in mRNA vaccines because they are stable and can produce a protein for upto 7 days. (ummm, isnt that highly codon optimized, chock full of N-1-methylpseudouridine, segmented poly (A) tail vax mRNA lasts long enough as it is?) This tells me that linear mRNA needs to be highly modified in order to make protein for long enough, whereas circRNA does not. Anyone?
Well, it is going to be the next big thing if we look at the what the Biotech companies are doing. Though this circRNA would likely be used to make therapeutic proteins, and not vaccines I think. Biotechs bet on circular mRNA
So what is the downside? Are they immunogenic like mRNA made via IVT is? Well, they can be immunologically silent AS LONG AS THEY ARE PURIFIED OF LINEAR RNA. Excuse me while I laugh.
Products of the circularization reaction include impurities, such as linear RNA precursors and spliced RNA sequences. The removal of linear RNA precursor molecules that had not been successfully circularized is a difficult problem faced by circRNA vaccine production due to the similarity of molecular weights between linear RNA precursors and the corresponding circRNA molecules. Research progress on circular mRNA
Isn’t this the same problem we have been dealing with? However, it looks like codon optimization, weird poly (A) tails and using modified nucleotides are not necessary. In my view, this is a tacit acknowledgement these synthetic modifications to mRNA cause adverse events (like frameshifting etc) which are especially a risk for repeated injections of mRNA products.
The adjuvanticity of the LNPs
Finally, the fact that mRNA needs an adjuvant is acknowledged. Because, the immungenicty of the mRNA is suppressed with the N-1-methylpseudouridine substitution, is it the LNPs that act as an adjuvant? If so, it is likely the ionizable component of the LNPs. However, it is probably both.
This brings up the question of how these lipids were studied and regulatory requirements. The lipids were reviewed as excipients (ie no pharmacological actions of their own), but in reality they are now acknowledged as adjuvants. A more thorough assessment would be required.
Liver and Spleen Toxicity
Potential liver toxicity was noted in the regulatory reviews of the EMA and others, but no real followup was requested. I am sure it got downplayed as these jabs were supposed to be given as 2 shots only. But repeated doses could certainly be a problem
I’m not sure if the authors in the Nature article suggest any mitigations on this toxicity
Immune Responses
Here, the authors of the paper do a great review of the types of immune responses occurring with the current mRNA-LNP platform
TLR activation and cytokine secretion
So, how can this be solved? Changing the immunogenicity of the LNPs? Finding new ones? How’s that for an adjuvant. Pfftt
Inflamasome actvation
This is a distinct innate immune system effect of the LNP-mRNA platform.
This effect also appears to be primarily from the LNPs themselves, because the mRNA was purified of dsRNA and synthesized using N-1-methylpseudouridine so was assumed to be immunologically silent. Hmmmmm, wonder if they considered lipid adducts as I discussed in previous substack? Too much residual DNA? Other contaminants? This shows us how little we actually know.
Complement activation and hypersensitivity activation
Finally, some attention is being given to CARPA and hypersensitivity. This too was identified as a potential problem in the regulatory review and should have been studied more. ALL NANOPARTICLES ARE RISKS FOR HYPERSENSITIVITY REACTIONS. This was known beforehand.
Ummmmm, did you read myocarditis as a result of HSR or CARPA???? That’s because a mouse study showed that inadvertent IV administration caused myopericarditis!! That’s because IV administration would make CARPA and hypersensitivity reactions worse than an IM administration (which could still be an issue). Seriously pegylated nanomedicines are known to do this, must be mitigated and are only given in life-threatening diseases. Complement activation is a very serious thing.
Understudied Areas of mRNA-LNP Toxicities
Particulate LNP-mRNA toxicity
I’ve been talking about how these products are nanoparticles and nanoparticles have specific toxicities.
Its the PHYSICAL properties of these nanoparticles that determines biodistribution and cell binding affinity and is specific for every mRNA-LNP combination. That is, it is possible that Moderna’s jabs is going to biodistribute and bind differently than Pfizers jabs. It is all about the biomodecular corona which needs another substack, lol.
Toxicodynamics of Extracellular mRNA-LNP
Extracellular mRNA-LNP particles? What are they talking about. These researchers claim only 1-2% of the jab results in actual transfection? Which means what happens to the other 98%? Again it is about organ toxicity and complement and other toxicities. Extracellular disintegration of the mRNA-LNP particles are also unknown. This is because NO STUDIES were done on the elimination of the mRNA, the lipids or the nanoparticles themselves. Finally they actually state that there may be issues with aldehydes and potential toxicities. All those lipid adducts running around which are now mutagenic. See my post Lipid adducts
Current Approaches for mRNA-LNP Risk Avoidance
What do they propose to mediate these serious adverse events?
LNP libraries or newer LNPs like those based on squaramide lipids (which Moderna has been working on). I don’t know, sythetic lipids are synthetic lipids, I think it’s gonna be hard to make them less toxic
Combination therapies. This would be for cancer only but they would combine the mRNA-LNP with, get this, a cytokine or an immune checkpoint modulator, which are pretty toxic by themselves. What are these people smoking? This is supposed to help? This is primarily for increased efficacy vs tumors. Or, alternatively they could just add dexamethasone or a steroid to the LNP to decrease the immune response (but without affecting mRNA translation, quite a trick). This also means that everyone should get acetaminophen, a steroid and Benadryl prior to any mRNA-LNP injection. This should have been the case for the vaccines but of course, the $$ and then this would cause vaccine hesitancy and show just how toxic these injections were.
Minimization of off-target effects. Uhhhh that would be great! Kinda late though. What do they recommend? Messing with MORE mRNA, now microRNAs? I mean, this could go both ways.
Another option is they could also add another cationic lipid to the 4 lipids in the LNPs so there is less hepatic targeting, and better targeting to the specific organ needing treatment. But didnt Pietr Cullis say he wore out 5 grad students trying to get specific on LNP biodistribution? I think this is wishful thinking. Plus synthetic lipids. Enough said.
New animal models
Organoids
Summary
I don’t think I can say it better than the authors:
As they said, these mRNA-LNPs are MORE THAN THE SUM OF THEIR PARTS. This is key. We think of them a separate entities, but when IVT mRNA is combined with these LNPs they then are a separate entity in, and of, themselves. You cannot just change a lipid, or the size of the mRNA etc. That changes the whole nanoparticle and with it, translation kinetics, biodistribution, toxicity etc.
However, the authors have done a great review on the toxicities and issues of the lipids and the mRNA-lipid nanoparticles. I just do not think their proposed solutions will actually work. The ongoing issues are really primarily
lack of targeted biodistribution
mRNA immunogenicity and purification
elimination of the mRNA-LNP particles
nanoparticle and nanomedicine intrinsic toxicities
Thanks for reading. And pray the rosary.
Brava. I can actually follow along! Thank you for making things crystal clear.
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