When Ana Moreno started her PhD in bioengineering at the University of California, San Diego, she wanted to design gene therapies for hard-to-treat diseases. But targeting the genome seemed like a heavy-handed solution for diseases that involved more nuanced changes in gene expression, such as chronic pain.
Instead, she used an approach called CRISPR-dead Cas9 (dCas9), which takes advantage of CRISPR’s ability to home in on a target gene. Once the CRISPR machinery gets there, dCas9 doesn’t make cuts. Instead, it tacks on molecules that either increase or decrease gene expression. In pain mouse models, this strategy reduced the expression of a gene encoding a sodium channel known to be overactive in chronic pain.1 After CRISPR treatment, the animals appeared to be in less pain.
Now, spun from her PhD research, she has founded a company called Navega Therapeutics to develop this pain-reduction strategy into an alternative to addictive opioids.
Why use CRISPR to control gene regulation?
When we think about common diseases and going beyond rare diseases, we need to utilize genome regulation. Genome editing cannot target more than one gene, and we get translocations or other problems. Some diseases require multi-gene targeting and multiplexed targeting—that is where we think this approach will be really exciting.
What makes dCas9 useful for this?
Traditionally, CRISPR uses a nuclease that makes double-stranded breaks. But in dCas9, these nucleases are mutated so that you have no more DNA-cutting activity. However, it still retains the guide RNA, this GPS of where it binds in the genome. We can design the guide to go where we want it to in the genome, and then add activation or repression domains to activate or repress genes of interest.
Specifically, when we’re looking at pain, we know that the gene that encodes NaV1.7 is a very plastic gene that increases expression all the time. The advantage of gene regulation with dCas9 is that you can downregulate that gene, but also prevent future upregulation.
How did you decide to use this strategy for pain?
We have all heard about the opioid epidemic. We know that there has not been an advance in developing drugs for chronic pain. It is a difficult disease to tackle because we know that opioids work well, so it is difficult to go beyond them.
It was one of these random Sunday nights reading papers when I came across NaV1.7. People that have a mutation in its gene—a loss-of-function mutation— might feel no pain, but there are also individuals that have a gain-of-function mutation that causes more pain than in normal individuals. I thought it was really exciting, especially because I was using dCas9, and thinking, what can we utilize this technology for? It made a lot of sense to have editing for pain.
What do you have to consider when you are working with a condition like pain that has a role in normal function?
When we are going after pain, we do not want to permanently mutate that gene. We want a solution that represses it but is not permanent. We are starting with inherited erythromelalgia, a rare disease where patients live with this gain-of-function NaV1.7 mutation. Treatment with dCas9 would be a cure for them. Then we can think about other neuropathic pain conditions that could benefit from gene therapy using the same drug product.
For my first time working on pain, I was excited and surprised that it works—in the sense that things work in vitro all the time but then in vivo there are a lot of issues—and that it not only prevents but also reverses pain, because that is what we want in the clinic.
What motivated you to start a company?
The motivation was being really passionate about the results and thinking, I do not want to stop at the mouse model, graduate, and move on to something else. I want to follow this into the clinic. We get a lot of emails from patients suffering from pain, which is very motivational. Someone really needs this. Sometimes science is luck too: you have the right lab, the right technology, the right application, and that drive. Right after my PhD, we started the company Navega Therapeutics and are at JLABS now, an incubator space from Janssen.
What do you do day-to-day?
Startups are like academia. We wear a lot of hats and do different things. It is like grad school: when equipment breaks, we have to figure out how to fix it.
When I started, it was bench work all the way. I took a picture of my first bench because I was excited that it was my bench. It is hard to find someone that is specialized in these different components, so I still do a lot of mouse work. But now we have scientists and research associates that are at the bench, so my days are more management-style where I feel more like a PI. In graduate school, it was research and publishing and reading papers, and now it is that plus everything else. I write grants, manage the lab, talk to investors, write patents, and think about regulation.
Sometimes my old PhD friends ask me if I get bored working on the same thing, but it is not the same thing. With other aspects of the business—IP, patents, lawyers—it is different and novel.
How did you learn the skills to lead a company?
We incorporated the company in 2018 when I was still working on my PhD. I tried to look for resources on campus. There are some incubators; some were focused on female founders, because there are different challenges for female founders. I ended up taking around a year of business classes and got a “mini MBA.”
A lot of it was trial and error. If you look at my first pitch deck and the one that I have now, it is very different. I think the biggest thing I learned is that, as scientists, we think everyone is going to want what we are producing or doing. But then we have to think about other aspects: who is our market audience, who is going to need our therapy, what is our pricing, what about reimbursement, who is going to manufacture this? All of these things that we do not have to think about when we are in graduate school.
I have been learning along the way from great advisors. I have met amazing people that have been really generous with their time. They help us grow the company and believe in what we are doing. It is good to know that there are people out there like that.
What's coming ahead in the next year?
We are at the preclinical stage now and are excited about our work that showed the concept. Now we are doing the IND-enabling studies to get this into the clinic and expand into other targets. The next steps are to grow the team and operations, and then hire people to think about quality control, quality assurance, and other components that are required to introduce a drug to the clinic.
This interview has been edited and condensed for clarity.
Reference
- A.M. Moreno et al., “Long-lasting analgesia via targeted in situ repression of NaV1.7 in mice,” Sci Transl Med, 13:eaay9056, 2021.
