Hydroxychloroquine blocks SARS-CoV-2 entry into the endocytic pathway in mammalian cell culture
https://www.nature.com/articles/s42003-022-03841-8
Abstract
Hydroxychloroquine (HCQ), a drug used to treat lupus and malaria, was proposed as a treatment for SARS-coronavirus-2 (SARS-CoV-2) infection, albeit with controversy. In vitro, HCQ effectively inhibits viral entry, but its use in the clinic has been hampered by conflicting results. A better understanding of HCQ’s mechanism of actions in vitro is needed. Recently, anesthetics were shown to disrupt ordered clusters of monosialotetrahexosylganglioside1 (GM1) lipid. These same lipid clusters recruit the SARS-CoV-2 surface receptor angiotensin converting enzyme 2 (ACE2) to endocytic lipids, away from phosphatidylinositol 4,5 bisphosphate (PIP2) clusters. Here we employed super-resolution imaging of cultured mammalian cells (VeroE6, A549, H1793, and HEK293T) to show HCQ directly perturbs clustering of ACE2 receptor with both endocytic lipids and PIP2 clusters. In elevated (high) cholesterol, HCQ moves ACE2 nanoscopic distances away from endocytic lipids. In cells with resting (low) cholesterol, ACE2 primarily associates with PIP2 clusters, and HCQ moves ACE2 away from PIP2 clusters—erythromycin has a similar effect. We conclude HCQ inhibits viral entry through two distinct mechanisms in high and low tissue cholesterol and does so prior to inhibiting cathepsin-L. HCQ clinical trials and animal studies will need to account for tissue cholesterol levels when evaluating dosing and efficacy.
This paper quoted by FM1 is interesting, and a great example of how science can be misread.
It shows strong evidence that HCQ is more effective against COVID-19 in cells in vitro with high cholesterol levels, and gives insight into how that efficacy works.
That is very helpful in evaluating when HCQ is more likely to work against COVID, or in increasing the efficacy of HCQ (by adding other drugs).
It says nothing about whether HCQ is actually effective in vivo!
How is it that these lab studies (not just sometimes - usually) - prove to be misleading?
For in vitro results to predict that a drug works you need:
(1) the drug can kill covid faster than covid reproduces
(2) the concentration needed to do that is safe
(3) the drug when administered gets to the right place - so that drug levels are high enough where they are needed.
(4) the in vitro culture correctly mimics what happens in the much more complex human body
All of 1,2,3,4 are needed for in Vitro studies to predict drug performance. 4 is particularly difficult, which is why in vivo studies are a starting point for further work but often do not predict drug performance well.
However, for in vivo studies to predict anything, you need to compare in vitro concentrations that kill the virus with in vivo safe levels.
Any study that does not at least mention whether the levels they use are tolerated by humans should be taken as meaning those levels are in fact not prcatical - or at least not known to be practical.
So this specific study - because it does not answer that key question - says nothing about whether HCQ might be expected to work in humans against COVID.
What actually works is always unknowable due to 4. Maybe in reality it works better, or worse, than expected.
The experience for anti-viral drugs is that it is very difficult to find new effective ones. I guess the human body is already pretty good at dealing with viruses.
Anyway - FM disagrees with me about what works because he reads and posts all these hopeful-sounding studies without checking whether hopeful-sounding is the same as evidence it works.
Since all scientists will hope their work might end up useful, and work that is interesting will get published regardless of is it useful, we get a lot of hopeful-sounding papers.
THH