Tell us about your current work. What does your PhD project involve? (Amy)
I am currently a PhD student under Borries Demeler. I have been working with him for about 2 and a half years now, and my PhD project is actually studying adeno-associated viruses (AAV), and trying to develop a couple of different methods so that we will be able to accurately determine the concentrations of each of the different components that can be found in an AAV formulation. The project I’m working on with API uses the same techniques as my PhD work, so in that way, they are connected.
What are you and Amy working on with API? (Dr. Demeler)
We are analyzing the nsp14 (non-structural protein) in COVID-19. That protein has never been expressed or purified before, so there are several students involved in this project including one from Dr. Trushar Patel’s group at the University of Lethbridge.
The idea is that this nsp14 protein is necessary to interact with a cellular protein called DDX1. DDX1 is a helicase that unravels double-stranded RNA so it can be copied and read, and made into additional viral RNA molecules. If we could interrupt this process, we could prevent the virus from replicating. Our biophysical and biochemical approach is to first purify these two proteins to a high degree. We can then stick them into our analytical ultracentrifuge and measure their interactions. Before we do that, we have to verify that they are pure so that we have controls. This is what Amy’s been working on. We also got a CFI grant to support this study, so we were able to buy a supercomputer, which is helping to support this Mitacs project with API as well. They are very complementary.
The goal is, once we understand the structure of these two proteins, and we understand where they interact, then we can find a drug molecule that would bind to this interface and inhibit the interaction. So that’s the whole idea, inhibit the interaction, and prevent the viral replication. A lot of drugs that you find today are working on this principle – by inhibiting a function that comes from the viral particle.
Tell us more about some of the exciting work that has been happening in your lab (Dr. Demeler).
There’s a very related effort (to the work with API) that Amy was involved with that was just published in a very high-ranking journal. Amy’s the first author on that. It was published in ACS Nano and is a collaboration with UBC, among others. We developed a methodology where we measure the density of molecules that are used for gene therapy like AAV, but also in this particular case, using lipid nanoparticles. Groups were traditionally measuring the structure of these particles by cryo-EM, but they could never tell the difference between a loaded or an empty lipid nanoparticle. This is now what Moderna and Pfizer are doing for their vaccines – they load the lipid nanoparticles with RNA, which is what we did. And we developed a method to measure how loaded the lipid nanoparticles are. We did this by analytical ultracentrifugation, because cryo-EM could not pick it up. That worked really well, and is actually a big deal.
What sort of broad applications could your recent work have in the life sciences industry? (Dr. Demeler)
If you look into gene therapies, it is an exploding area of interest. Especially in the area we are working, analytical ultracentrifugation, it seems to be the gold standard for characterizing these gene therapy formulations. They all share the following idea – you have a vector like a lipid nanoparticle or viral vector capsid, which can be loaded with a drug. And then you inject that into a person and the nanoparticle will find its target, like a cancer cell, or maybe it’s a blood cell that generates sickle cell anemia that needs to have a fix for the hemoglobin gene so it doesn’t produce sickle cells anymore. But, before you inject something into a patient, you need to be certain you don’t have contaminants in there and that the particles are properly filled, not overfilled or underfilled.
This is something you can do with our technique that Amy is an expert in. This applies to pretty much anything that’s being done in gene therapy, whether that’s lipid nanoparticles, or AAV, or any other vector that can be used to inject a cancer drug, or antiviral, or vaccine, any one of these can be used this way. For the biopharma industry, I would say it is a very high priority to start using our technique for characterizing drugs before they get injected into people. We’re working with the manufacturer of our instrument to get the FDA on board to develop good manufacturing practices (GMP) that will be followed by the manufacturers, and used to validate drugs before they get approved.
What has it been like working with Dr. Demeler and collaborating with many different researchers in Alberta, BC and beyond? How has that been important to your development? (Amy)
It’s been such a great experience. I’m so happy that I chose to work with Dr. Demeler – he’s very knowledgeable about his field, and about many others as well which is really helpful. And as you can tell, he has quite a passion for this work, so it’s hard not to have that rub off on you. He also has connections all over the world. We’ve been working with companies in the states, the UK, and more. It’s really great to be able to meet all of these different people, and to make such big connections. We’re learning about things outside of our own field, and it’s been exciting to learn about the passions that others have too.
API funded me going to Caltech to learn about working with AAV for the first time. It was such a great experience, everyone was so wonderful and we’re still in contact – they are helping me when I have questions. I visited the CLOVER Center run by Dr. Viviana Gradinaru. I spent 10 days there and we went through the whole process of making adeno-associated viruses twice. They answered all of my questions. We hadn’t set anything up like that here at the University of Lethbridge yet, so it was a great learning experience. Having to learn all of that on my own would have been a bit overwhelming, so it was nice to have somebody walk me through the steps.
Why do you think it’s important to mentor the next generation of researchers and scientists? (Dr. Demeler)
I am kind of in the end-reaches of my career, and I could probably retire in a few years, but, when you spend your entire career developing a technique or a method, you have a lot of fun along the way. Every other day you have some sort of eureka moment and it’s very fulfilling, but at some point you get to the stage where you say – what if nobody can take this over? And I did this all for nothing? And nobody knows how to do it when I’m gone? So you have to either clone yourself, which is not really feasible, or you train the next generation which is the next best thing. When you have some fantastic students like Amy, this is just thoroughly a fun thing to do. To find somebody that has the smarts to pick this up, and then to do their own thing with it and run with it, come up with new projects, for example the AAV project, Amy came up with that on her own. I’m finally at a University where I have access to undergraduates and graduate students that can work in my lab full time, and I’m just repeating that joy.
What would be your top piece of advice for students interested in becoming a researcher like you? (Amy)
Do some research before you commit to anything so that you’re working on something you’re interested in. The work goes by a lot faster and a lot better if you like it. And start early. At the University of Lethbridge, you can get into labs in your undergrad, so you will know what to expect and you can start talking to the PI’s. All the PI’s here are really friendly so you can talk to them to see what work you find the most interesting. And one thing is to know what you want. You can find someone you want to work with, and then you can tweak the project to fit what you’re interested in. Follow your passions. Do what you want.
How important has collaboration been as part of your projects, and what have you learned from collaborating with others within Alberta, Canada or around the world? (Dr. Demeler)
This is absolutely critical, crucial, essential. Without collaborations, we are nothing. Science has, over the last 10, 15, maybe even 50 years advanced to such a degree that you can’t be an expert in everything. It’s impossible. Unless we work with collaborators, we aren’t going to be nearly as successful as we are now. And, if you look at the people that have collaborated with the Canadian Centre for Hydrodynamics over the last 2 and a half years, since I moved to Canada, we already have over 90 people that we’ve collaborated with from all over the place. You put all of these groups together and you can solve real problems. That’s what we’re all about here, it’s exciting!
About the Interviewees:
Dr. Borries Demeler is a Professor in the Department of Chemistry & Biochemistry at The University of Lethbridge, a Canada 150 Research Chair for Biophysics, and Director of the Canadian Center for Hydrodynamics. For more information on his work, please visit: https://www.demeler.uleth.ca/
Amy Henrickson is a PhD candidate working in Dr. Demeler’s lab. For more information on her work, please visit: https://uniweb.uleth.ca/members/1232/profile