A quick summary and a final evident clue

"It's just a bit cholesterols"

Hopefully you remember my most monstrous substack, where I tried to explain to you many of my learning sources and basic understandings of signal transduction pathways and their crosstalks (mutual influence) and the catastrophe of LNPs?:

Do you remember the narrative: “PEGylated lipids, cholesterol, phosphorylated lipids and cationic lipids?”

Too bad it's not “just” cholesterol. And with these papers, from which I would like to quote briefly, the final proof has been given that it has inevitably affected the PtDins-cycle that I so hotly discussed and thus consequently all the processes linked to them:

Let's see what they wrote when they presented and later reviewed their silly idea of LNP-Tech?:

Lipid nanoparticles in the development of mRNA vaccines for COVID-19

“Structural modification of cholesterol derivatives increased the cellular uptake of LNPs and trafficking by 25-fold [55]. “

Please keep this source in mind cause I’ll show you the shit show behind synthetic cholesterols in a few seconds. I just wanna make clear: They did not use natural cholesterol:

Effect of mRNA-LNP components of two globally-marketed COVID-19 vaccines on efficacy and stability

Let’s look into this if it’s the same as discribed official:

https://www.biochempeg.com/product/mPEG-N,N-Ditetradecylacetamide.html

https://www.biochempeg.com/product/Cholesterol-(Plant-Derived).html

https://www.krackeler.com/catalog/sigma/SIGMA/C8667

CAS number fits, does it?

And now it’s getting nasty: Remember the mentioned Source 55?:

Naturally-occurring cholesterol analogues in lipid nanoparticles induce polymorphic shape and enhance intracellular delivery of mRNA

Let me quote:

"This enhanced fragility, originating from an altered surface composition and shape, perhaps promotes eLNP fusion with the endosomal membrane. It is possible that eLNP shape and surface lipid composition leads to differential trafficking that drives it to privileged pathways more suitable for escape from the endosomes. It is probable that higher retention of eLNPs due to a decrease in its interactions with lysosomal transporters provides them enough residence time to escape from the endosomes subsequently causing enhanced nucleic acid delivery. Whether the structural characteristics, surface lipid content, and interactions with the cells can improve gene delivery in vivo remains to be explored. Understanding the nanoparticle design, controlling its structure and its surface lipid composition can enable non-viral vectors to breach this most formidable intracellular barrier—the endosome."

"These findings suggest that C-24 alkyl-substituted cholesterol analogs can boost transfection independent of cell type, ionizable lipid, or nucleic acid cargo."

So to think it through: let's assume for a second that the phospholipid bilayer is not directly penetrated, as some really stupid papers are telling in using words like “fusion” and “endocytosis”. But the PtDins cycle is a 3D gear that regulates bilayer structure - endosomal-lysosomal trafficking. So they write with this little sentence:

“This increased fragility, due to altered surface composition and shape, potentially promotes fusion of eLNP with the endosomal membrane.”

Bye, bye PtDins cycle.

My point is that they are clearly describing endosome fragility and increased leakage from endosomes, r8? Consequently, this will have a massive impact on the PtDins cycle (we are talking about a permanent cycle of different chemical transitions). Consequently, disruption of phospholipid reorganization is inevitable.

And don't forget the fun with cationic lipids, which I seriously doubt phosphatases like SHIP1/2 and PTEN can handle.

”The diminished membrane ordering capability of C-24 alkyl derivatives of cholesterol is well documented^28,31,66,67.”

And the really crucial point here is that the mechanism behind it is described. So it is not so important whether it is modified as a derivative with a 25-fold increase or not.

Addendum:

As far as the manufacturing process is concerned, extraction from plants is not profitable. So is it synthesized from phytosterols? Or synthesized from deer, as shown in the CAT number example. The problem of isomers and impurities is already pre-programmed. Even with 99 percent. Help me out and dilute to 1 per hundred (one percent) impure phytosterol residues and isomers: what was that about parts per million? And it gets even better: no one can tell you whether they have changed the cholesterol or not. Because the CAS number doesn't tell you. Not even the degree of purity is known.

See also:

https://www.lgcstandards.com/DE/en/Cholesterol/p/TRC-C432501?utm_campaign=LGC_(Generic)_Germany_English_(TRC%20Product)&utm_medium=cpc&utm_source=google&gad_source=1&gclid=CjwKCAiA2cu9BhBhEiwAft6IxEa0kQs7DewyugKH75DEyEgXDNPyHEubzAc-QSDPJyf-c-MSM0_aoRoC8FQQAvD_BwE

That would also explain the randomized transfection efficiency even with the same cell types, wouldn't it?

Narfastic fun with your mTORC crosstalks and thus lipid rafts / receptor and ion channel functionality.