A short digression into COVID19

A month or so before the COVID19 outbreak really took off, I was visiting friends in Germany and we were discussing the virus emerging from Wuhan. We realised it had the potential to become a pandemic and I joked we might have to refocus our research onto COVID19. Two months later, it became apparent that the nanobit technology I had been using for cancer research could be harnessed to look for drugs to fight COVID19. I had been using "nanobit technology" to study a biological process called autophagy because I believe drugs which affect autophagy may treat cancer.

"Nanobit technology" is based on an protein in fireflies that allows them to give off flashes of light. The protein is called luciferase. In the nanobit system, the luciferase protein is split into two parts, so it does work and give off light unless you bring the two bits together somehow. If you put the two bits of luciferase onto two proteins that normally stick to one another, that brings the two bits of luciferase together too, and you can measure the light emitted.

It turned out that when the COVID19 virus gets into our cells, it binds to a protein called ACE2. And ACE2 forms a "dimer" - meaning two molecules of ACE2 stick to one another (called "dimerization"). I realised that I could use the nanobit technology to measure the ACE2 proteins binding each other. I wondered if the dimerization might affect the virus binding to ACE2 and entering our cells. And if so, could I find a drug that affected dimerization? Importantly, Keele University were kind enough to fund my research so I didn't have to wait months to get the funds through normal routes to test my idea. And I was happy to do the lab work.

So I attached the two bits of luciferase to ACE2 molecules, the ACE2 brought two bits on nanobit together and I got light emitted. Then a colleague from Birmingham, Dr Farhat Khanim, provided me with a collection of medical drugs. The drugs were all already in clinical use for treating various diseases. This meant alot was already known about using them in patients, so they could potentially be "repurposed" to treat COVID19. This is a lot quicker than trying to find a new drug from scratch, which can take years, if not decades. We didn't have time for that. Because of the COVID19 restrictions, I picked up the drugs from her in a car park, true drug dealer style. Back in the lab at Keele, I tested all 90 drugs and found a few that seemed to affect ACE2 dimerization. Clofibrate was the most promising, but this drug had been withdrawn from use because of nasty side effects. But there were other related "fibrate" drugs still in use. So I purchased those and tested them. It turned out that one called fenofibrate was the most promising and reproducibly seemed to increase ACE2 dimerization in the nanobit experiments.

Then other colleagues working at Keele, Courtney Mycroft-West and Mark Skidmore suggested they test fenofibrate to see if it had any effect on the viral spike protein. The Spike protein sticks out of the virus, and is the part of the virus that sticks to ACE2, allowing the virus to get inside our cells. They found that fenofibrate made the the spike protein less stable and that it also prevented the spike protein from binding ACE2. This was great news because the drug could potentially work in two different ways. But now we needed to know if it really altered the ability of the virus to infect cells.

Back to Birmingham. Farhat had been busy setting up tests with the live virus to see if she could find drugs to block infection. Farhat's tests looked at the appearance of a viral protein inside cells, after they were exposed to the virus. If fenofibrate worked, it should block that. At the same time, Elisa Vicenzi in Milan, with whom Mark was already collaborating, had a different type of test to measure viral infection called a "plaque assay" - this basically detects a colony of cells infected with virus. So this was great - two seperate labs could test fenofibrate at the same time.

I'll be honest, I wasn't really expecting it to work. We had used quite high concentrations of the drug, the effect on dimerization was modest, and other experiments I conducted made me worry the drug was doing something other than dimerization to have its affect in the nanobit assay. And just because the drug destabilized the spike protein, that didn't necessarily mean it would prevent infection, even though the drug reduced the spike protein from binding ACE2. But it worked in both labs. Farhat spent most of the Christmas vacation working on it and the Elisa's group got similar results at the same time. This was very encouraging because one of the best ways to confirm your results when you are doing science is when someone else can get the same results in a different lab.

We put the results out prior to peer review so the medical community could use the information to treat patients if they wished. Another group, led by Koby Nahmias in Israel had also suggested fenofibrate might be useful to treat COVID19 infection, although they thought it worked in a different way. This has led to some clinical trials of fenofibrate and their reports on social media sound encouraging. I am looking forward to hearing how they go. Hopefully we will soon see trials in the UK too, perhaps in patients prior to hospitalization.

After peer review the paper was finally published today here https://doi.org/10.3389/fphar.2021.660490

I don't know if it will make a difference, clinical trials will tell us that, but I have done what I can.