Thanks for talking to me Julia. You are currently doing your PhD in Biological & Biomedical Engineering with Amine Kamen. What academic and research experience did you have before you came to McGill?
I completed my Bachelor’s in Biotechnology in Brazil, where I’m from. At that time, I did a summer internship exchange here in Montreal and that’s when I knew I wanted to come back for grad school – I got a preview of Montreal summer! I started looking into different labs and came across Amine’s group and was very interested in their work on viral vaccines and gene therapy. I was interested in taking my biological knowledge and applying it to develop manufacturing processes for therapeutics.
Did you feel like you were missing any engineering knowledge when you came back to McGill for grad school?
Even now, sometimes I wonder how to define myself or my research background when I talk to people. I definitely had more of a biology background and I didn’t have strong experience in calculus or coding from my undergrad, for example. I had worked in a yeast lab developing genetic engineering tools, but not on scale-up or with bioreactors. But I feel I have always had the engineering mindset of “how do we achieve our objective with the tools we have?”. During my Master’s research, I learned so much about process development and, during my PhD, I have deepened my knowledge both from the engineering and biochemistry sides. I’m glad I have had these diverse experiences, as it helps me communicate with scientists and engineers from different disciplines and stages of drug development.
As a member of Amine’s lab, you have been very involved in D2R’s training activities, attending both D2R Symposia and several events. How do you feel you have benefited from being part of D2R?
I first heard about D2R through Amine, and our lab has shifted towards mRNA research since the Initiative started. Through being one of the first students in the lab working on mRNA, I have been able to learn so much by collaborating with professors across McGill. You can only learn so much yourself through reading papers. Within my department, there are not many professors working in this field - I have really been able to find my community within D2R.
One project you are working on is the “End to end sequencing platform for RNA therapeutics quality control” in collaboration with Professor Ioannis Ragoussis. What is your role on this project?
Whenever you produce an mRNA vaccine or therapeutic, there are very strict quality control standards. There are many things you must characterize to ensure that the therapeutic will be safe and effective. The way it is done now, there are several different assays to perform. However, it would be much more straightforward to streamline this process, and that is where nanopore sequencing comes in. Prof. Ragoussis’ group at the Genome Centre has been working on developing nanopore strategies to directly sequence the RNA molecule without having to make a DNA copy first. This shows promise to not only analyze the sequence of the synthesized mRNA but also to look at other qualities that we typically require other assays for, such as detecting the 5’ cap, measuring the length of the poly(A) tail, or evaluating RNA degradation. In this project, our lab’s role is to validate the new nanopore strategies through performing the traditional established assays and to generate samples from all steps of the process, like before purification, after purification, after encapsulation, etc. It’s myself and a research assistant in our lab, Laurence, working on that.
When did your lab begin working on mRNA therapeutics?
Since Amine’s lab was already working with vaccine manufacturing, we were keeping a close eye on the success of mRNA vaccines during the pandemic and how fast they were developed. I don’t want to call it simple, because it isn’t, but there is definitely less complexity in the steps required to make an mRNA vaccine compared to virus manufacturing that enables the development of a sort of plug and play platform.
Can you briefly explain these steps that are involved in mRNA therapeutic manufacturing?
Yes. It actually starts with DNA. First, we produce a DNA molecule that will encode for the desired RNA sequence. And that's typically done with bacteria - we transform the bacteria with plasmid DNA and as the bacteria grows and multiplies, it also replicates that genetic material, so that we can extract it from the bacteria at a much larger amount. There are also strategies now that enable doing that step without bacteria through enzymatic pathways. Once we obtain the DNA, we perform the actual reaction to produce the RNA, which is called in vitro transcription. This step, and all future steps, are cell-free processes - that's what makes it simpler than virus manufacturing that requires cell culture. For in vitro transcription, we add the DNA with the enzyme which will read the DNA and incorporate the correct nucleotides to synthesize the target RNA. The mRNA is then purified and, in the last step, is encapsulated into lipid nanoparticles with a formulation designed to preserve the particle for as long as possible.
Great, thanks! In addition to your work on the D2R-funded end-to-end sequencing project, you are also involved in the PATH RNA Cooperative project funded by the Gates Foundation. What is your lab’s role in this project?
In this project, several D2R-supported platforms are part of an RNA cooperative built to support core institutes in Senegal, South Africa, and Indonesia. Our lab’s role is related to developing and providing biomanufacturing protocols to researchers at these institutes in a technology transfer capacity. These include things like synthetic DNA amplification or protocols for RNA production and LNP encapsulation. I am working on these protocols through comparing different reagents and optimizing the steps involved. The goal is for these partner institutes to be self-sufficient in their production of any RNA therapeutic they would require. Additionally, the partners have a lot of information that we don’t – for example, in Senegal at the Institut Pasteur de Dakar, they have a lot of disease surveillance information that we don’t here at McGill.
It really seems like you are gaining great experience not only performing research, but also working on these collaborative projects. Does that influence what you envision for your future career?
Yes, I am coming towards the end of my PhD, so I have thought a lot about what comes next. The more contact I have with these types of projects, the more interested I become to move into industry as my next step to be involved directly with the manufacturing and development of RNA therapeutics!
That’s great! One of D2R’s objectives is to help develop a Canadian workforce in the RNA therapeutics space. Is there anything about yourself outside of the lab that you want the D2R community to know?
Outside of research, there are two main things I spend my time doing. Since moving to Montreal, I’ve been volunteering with a rescue group to foster cats. I still do that on and off because I ended up adopting one of the rescues so I can’t always be bringing in new cats without her getting annoyed! Also, I have taken up crocheting and knitting during my PhD, and it has become quite a serious hobby. I find it so satisfying to have a little project I can think about and complete. It’s good to have smaller projects to succeed on in parallel to the challenges of a PhD!
This conversation with D2R Training Program Officer, Anthony Van Kessel, has been edited for length and clarity.
