Hi, I'm Doctor Andrea Beal and today I'm going to be talking about my favourite topic, epigenetics and animal health. I'll present some applications and some emerging opportunities in this new field, coming to animal health. So, to start, I'll give you a little bit of background about myself.
I'm currently a postdoc at Nova Southeastern University. I'm also the founder and CEO of EpiPaws, an epigenetic testing company for dogs and cats. I've been studying DNA and gene expression since I was an undergrad.
So, during my time at Tarleton State University, in my bachelor's, I studied population genetics in the cotton rat. Then I went on to Texas Christian University, TCU to study RNA and gene expression in pipefish, very closely related to seahorses. And then during my PhD, I went to Florida International University, where I started studying environmental epigenetics.
I was looking at these epigenetic markers in sharks and dolphins. During this time, I created an age estimation test for dolphins, which inspired me to make age estimation tests for all different animals. And I landed in the pet space, wanting to do an age test for dogs and cats.
So I started Epipaws. . So today, we will have, we'll go through a few different topics and learning objectives, starting with some background on epigenetics.
You'll walk away with a better understanding of what epigenetics is and why it's important for animal health. We'll move into looking at DNA methylation, often abbreviated DNAM like this, for age estimation and how that could be useful in the clinic. We'll talk about an extension of this, which would be ageing rates and how we can use this for longevity and health monitoring.
And then we'll talk about the state of diagnostics when it comes to this kind of technology and wrap up everything by how we can implement epigenetic testing in the clinic. So diving right in on the background of epigenetics and DNA methylation, I'm gonna take everyone back to their sophomore year in college where they were learning about about genetics and how every cell in the body contains the entire genetic code. That whole entire blueprint is in all of your cells.
So then, how does a liver cell know how to act like a liver cell and a muscle cell know how to act like a muscle cell? Well, that's where epigenetics comes in. Epigeneex was first discovered and studied in the context of how stem cells, when a baby is formed, how the cells differentiate to form all these different tissues.
Since then, we've realised that these epigenetic changes continue to adapt and change inside a growing, animal. To help prepare them for everything they're going to encounter, and so the definition has changed to the study of heritable changes in gene expression that occur without alter alterating that underlying DNA sequence. Essentially, we're studying modifications to DNA that result in genes being turned on and off.
And why this is important in health is because of a few key features we find with these epigenetics. One, they're reversible. Any change we see, we could reverse it back, especially if it's a bad change we don't like.
These markers are highly affected by environmental lifestyle, environment and lifestyle. So everything from diet, stress, toxins, changes in temperature, seasons, all of that is affecting these markers and helping an organism adapt to the changes. So it can also help us in the context of health, understand how these things are affecting health.
They also can sometimes be inherited. Across cell divisions, so within an organism that's just making new cells, and then also across generations, so being passed on to subsequent generations. So lots of health implications there.
We're going to focus on DNA methylation, which is a modification that happens directly to the DNA over here. But I slipped this slide in just to mention a couple others that you might start to hear about. So one is histone modifications.
Histones are the proteins that DNA wraps around when we package up chromosomes, non-coding RNAs such as micro. RNAs, which are really coming out into the field for diagnostics. This is another type of epigenetic mechanism that can stop the process of gene expression at a different location than where DNA methylation functions.
So, like I said, focusing on DNA methylation, which is the most studied epigenetic mechanism. It's a mole, it's a methyl group attached to the outside of cytosine, which is the base in, DNA. You might hear me say CPG site.
This is a cytosine next to a guanine, very common area for this to happen. But don't worry about that. We're gonna really focus on the key reasons we wanna know about DNA methylation when we're talking about animal health.
First is that changes with environment. Let's look at some examples. A classic example is the GUI gene.
So, in mice, we see that there are different phenotypes showing here, brown mouse or yellow mouse. The brown is the normal phenotype. Yellow is the abnormal where we have hypomethylation.
So less methyl groups at the Agui gene. And it results in a yellow mouse, but they're also often obese. So health implications there.
This was connected with the diet of the mother. So BPA exposure caused hypomethylation and therefore the yellow obese phenotype. So, implications of diet on DNA methylation.
Here is a study that this is for my PhD. I'll point out the big takeaways from this, but I was studying baby lemon sharks at Bimini. And we have a northern nursery, a southern nursery.
And then in the early 2000s, there was a major dredging event. So what happened here is that the Bimini shark lab, they were sampling these sharks in these two nurseries during this time and archiving a piece of fin clip. So I came in and wanted to look at, well, how does DNA methylation change with this dredging event.
And what this lab noted was that during the dredging event, there was a decrease in survivorship. Of lemon sharks in this northern nursery, and they thought it to be because metals get introduced into the environment and get into the fish, and the fish are the food of the sharks, so it's a dietary issue going on for that survivorship. So what we found is that, and I'll walk you through this, the before north.
Was grouping really closely together and here's our caveat. We don't have before South samples, so our data set isn't as complete as we would like. And then we have during north over here, during that dredging event, being much different from during south, and then here is also after north, after south nurseries.
So, Looking at this panel, this is as if we're looking across this plain. We do see the during North having a very different DNA methylation pattern than the other groups. And so here are the takeaways I want you to realise here.
One, our data set isn't as complete as we would like. That's a baseline issue. And baselines are very important when we talk about animal health.
So I'm gonna bring this back up later. We'd love to have samples to have a better picture here. 2, things happen very quickly.
So we do see a change in the DNA methylation pattern very quickly. As soon as that dredging event, I know we had time periods, but all of them are grouping very, very tightly here. It's really fast that we see these changes happening.
So, moving right along. Now, getting into age estimation, Here, age estimation was for using DNA methylation was first pursued in humans. Now, why would we want to know age estimation in humans?
We typically know birthdays, right? So this was for forensic purposes is what they really honed in on. So if there's a crime scene and there's blood, that's all you have to go off of.
You can now look at, of course, we can see if it's male or female blood. And now we can see, well, how old are they? So, scientists were noticing that DNA methylation changed along with age, and there was a correlation, and this is across several different genes.
And we can make a model out of this for really accurate age prediction. From this, during my PhD, I looked at human studies. I looked at whale.
There was a whale study that came out, and I was able to create an age estimation test for bottlenose dolphins. It was the very first one for dolphins. Since this time when I translated it for dolphins, there's been other studies come out that have further improved this.
I kind of cherry picked my, gene targets in my model, but, I'm still validation that we can do this with these markers. From there, I wanted to make a destination test for all different animals. And so that's where I landed on doing one for dogs and cats and starting my company Epipaws.
So, I developed an age estimation test done from a mouth swab of DNA so it'd be really easy to sample and, and do the test. The accuracy of this test is plus or minus 12 months. And most, 75% of validation samples came within 5 months of actual age.
So we really recommend this for adult animals. There's also a difference in how DNA methylation changes in a puppy versus an adult. So we really honed in this test to be for adults, .
And animals that have more than 12 months of uncertainty around that age is probably where you're gonna get the most impact, but it does provide a really good feedback on, are we in the right ballpark? Was this estimate from a shelter correct or not? Things like that is where it really becomes helpful.
And then also in determining what kind of care this animal should get, going forward, age is a big factor in that. All right. Moving on to an extension of age estimation that's more applicable to health would be the ageing rate.
Here, we have chronological age, which was the test we just looked at, which is birthday, when we want to really accurately understand how physically old they are. Then there's biological age, which is the internal ageing rate of cells, and how healthy an individual looks or is, is the feedback we get. So, as of right now, age is the number one predictor of disease development, and this is chronological age.
But we know that 2 11-year-old dogs or cats might not have the same health and the same risk of disease development depending on lifestyle factors and how they've been treated their whole life. So biological age really provides a better insight into the health of a particular animal and their actual health or risk of developing disease. So how can we measure this biological age?
There's actually many ways and many molecules. First would be traditional blood chemistries. So, in particular, Purina just came out with a paper that they published where they created a biological age, model based off of blood chemistries.
Then there's phenotypic measures. This can be anything from just physical features like wrinkles in humans and things like that, or other phenotypic features that change with age, and if it's happening prematurely faster, we can calculate that into a biological age. Proteins are, are really great measure of disease risk actually, and we're seeing this in humans.
The problem with using proteins in the body is that it's really expensive to do this kind of analysis. So we haven't seen this take off really. It's great for R&D though.
And then there's epigenetic measures such as DNA methylation, which is of course what we're going to stay and focus on. So in humans, biological age testing is exploding. If you Google bottle biological age test, you don't even put in human, you're gonna see a bunch of human tests that are on the market for sale.
They range the whole gamut from blood chemistries, proteins, you can do proteins, DNA methylate. All of that's out there. One in particular that you may be familiar with is Function Health.
They're doing a really great job. They do all kinds, over 100 labs. They also have DNA methylation tests coming out.
And there's, I know our vet advisor, she uses this. And so, we've been really talking about this and how well they're doing in the human space, providing this kind of technology. For health monitoring and how can we bring this to pets.
So I think that this is really the future of proactive care for our animals as well, just like we're seeing in humans. But again, focusing back on just DNA methylation for biological age clocks in humans, this is what exists for them right now, is these 4 iconic, there's more than this, but these are the 4 iconic biological age clocks available. The two on the left here, Hannam and Horvath, these are trained on chronological age.
They don't take into account health factors. So you could say they're chronological age clocks, but they're using so many markers that there is a little bit of feedback and influence from health factors on these age estimates here. That would be a major difference I see between this and like the chronological age clock that we created, that's really specific for chronological age for our pets, and even the forensic, chronological age clocks they use for forensics are different from these.
So, and we have a tissue-specific one. We have a multi-tissue one, that's their big differentiation, I would say. And then over here on the right, we're seeing pheno age and grim age.
These two did take into account health factors and in particular, pheno age takes into account clinical markers, blood work. In, in other measures, and it's really good at seeing morbidity and mortality risk. And then Grimmage is very sensitive to lifestyle factors and being able to get a feedback on is this food better or this food better for this person.
. And so with these, if we're thinking about translating these to animal health, we definitely want to be in this realm here if we're talking about biological age and how we intend to use that. So, at Epipause, we have been working on this, and there are other companies as well working on biological age. And a big takeaway here is that you can make a million different biological age clocks.
They'd each tell you something different. It's the markers you put in. So, here, we are developing one that we want it to be sensitive to disease risk.
Is there a disease developing? We want that kind of feedback. And then we also want to be sensitive to lifestyle.
So, Those are the markers we're trying to hone in and like kind of data set that we're trying to train these models on is so that we can pick up on how is the overall health over time, and this is how we intend to use this is measuring it over and over again. So we need a nice baseline, so it'll be sensitive to see changes. And then, We can catch things like this, like if there is a change in diet.
Does it really significantly reduce their biological age and how they look from a health standpoint? And so this is coming out soon for, pet owners and veterinarians to start using. Right now, we've been doing research behind the scenes with nutrition companies.
So, Looking into that a little bit about biological age for understanding diet, there was a study in humans called the Sisters study, which looked at 50 over 50,000 women on different diets, and they did see an inverse, correlation in lower biological age scores for individual for cohorts that were on better diet. And notably, they made a note that it seemed to be very impactful that someone was on a good diet if they weren't getting proper exercise. So a good diet can go a long way if you're not as active as you'd like to be.
I think the same will stand for our pets as well. So, currently, what's going on for dogs is we're starting to see these biological age models be created, and then they're being tested on things like fresh food versus kibble, which is a study that we have going on right now, and we should have published early next year, . So we can start digging into how can we use biological age as a feedback on if our pets are on the right food.
So, this is a fun one here. This is not here yet for pets. It's what we're working towards.
But right now, we're at a state where we can assign a biological age based on these markers. But there's also a way to differentiate and understand, well, if we see an accelerated age, so an older biological age estimate than what the pet is. What is it about that pet that is unhealthy?
What's off track? How can we get it back on track? I think one of the best ways to do that is to understand the health of each individual organ.
So here, we're showing, let's say, it's a 10 year old cat. And so we would see here that the heart looks healthy, liver not too bad, stomach looks good, but then kidney. Here, the kidney.
Is ageing quickly. So, Jody's kidney score is 12, that's 2 years older than expected. This is where we would suggest to a pet owner to go visit the veterinarian, run some tests, and talk about how can you support the kidneys better.
Is it hydration? Is it something we need to change, lifestyle, anything the vet knows could help contribute to better kidney health. And getting ahead of things like chronic kidney disease that often pop pops up later in life.
We're studying this in not only mouth swabs, but we're also doing some work in blood. We wanna start doing things in urine just so we can create also really sensitive diagnostic tests from these markers as well that help us pick up on individual organ health. OK.
So, that brings us right into, last but not least, the early disease detection, which is an extension of health monitoring that we can do with these molecules. Oh, human epigenetic diagnostics, we have a few in, out in the market already. One is Cologuard.
You've probably heard of Cologuard. So, this detects DNA methylation in a faecal sample as well as some other markers that get measured from that faecal sample to help screen for colon cancer. Then we have the gallery test, which is really cool.
This one screens for 50+ different types of cancer. And it's measured from cell-free DNA from a blood sample. It's a screening test.
And then we have Of EpiSign complete, which is screening for different diseases, or actually, it's diagnosing diseases. This is a true diagnostic in the human space. And this is used after genetic analysis can't help us understand a rare disease that is occurring and we're not sure what it is.
Sometimes DNA methylation can bring to light what that is. So these individuals can get some care. This includes some really rare diseases like Kabuki syndrome.
X-link disorders, things like that, that you may or may not have even heard of. So, now, I'm gonna talk about a couple of diagnostics that I think will translate to the pet space really quickly. We're working on some of those ourselves and be really impactful.
One is chronic kidney disease. What I'm showing here is scientific articles in humans. So there's a lot of promise here for really early diagnostics and pinpointing new therapeutic targets.
For chronic kidney disease when it comes to humans, and I think a lot of this will be translatable. We're understanding in particular this last one, that DNA methylation is playing a role in the development of chronic kidney disease. There's a trigger that starts DNA methylation change.
And it just keeps going and it contributes to the development of disease, which was really interesting and something that we're exploring to see if this is the same that we see in dogs. Is it the same that we see in cats. So lots of potential here and I do think that possibly within a year or two, we'll have a diagnostic based around DNA methylation.
Maybe it will stand alone, maybe it'll be in conjunction with other markers, to diagnose this disease earlier. Osteoarthritis is the other one. And with this one, when it comes to humans, we're seeing DNA methylation markers that could serve as again, therapeutic targets, which is great, but also telling us more about the status of the disease.
And I think when we think about dogs and cats, this can be really impactful because They can't exactly tell us when they're starting to feel the very first signs of this disease creep in. And it's often not until they're really showing signs of pain that we're diagnosing this disease. So they're suffering unnecessarily almost.
Unless we come up with a really sensitive diagnostic for this. So I think this is a really, really cool one that has a lot of promise to come out in the next few years. So, now, we'll just move into wrapping this up with how can we start implementing and thinking about using Epigeneex in the vet clinic.
A lot of it's still to come. We have a lot of research going on, but that's something else you could get involved with. So, quick recap what's available, and I know I'm being biassed.
I'm showing EpiPaw tests. There are other, companies doing DNA methylation testing for dogs. I haven't seen another one for cats yet, so we're currently the only one for cats.
But there are other tests out there that do age chronological age estimation. I'm starting to see biological age come out, which I think that's really interesting cause we could end up with different, we will end up, we will end up with different biological age tests that might be better for one use case versus another, but we'll cover more of it. So that's amazing.
So with age estimation, we'll talk about a few ways you can implement that and then biological age and health monitoring. Very important. This is something that we want to build up a baseline, keep that going.
So how can we implement this in the clinic? One, starting with that chronological age estimation, the new, newly adopted dog or cat. I'm sure you've had that question.
We got him from the shelter. They said this age. We're not so sure.
What do you think? And we're going off of physical features, which can be challenging. This could be a nice internal molecular feedback, you know, another tool to use cause there is that error around the estimate, but it can really pinpoint, around the age that they are.
With that, it can be helpful if you're getting that age figured out to developing that health care plan, how often to test and screen for diseases, how often to do vaccines. There's all kinds of health care plan, things around age that you could need or want that age estimate feedback on. Especially if you're starting to see a disease develop.
Are they really that old? Are they not that old? What's going on?
With this, this is where we also transition into how can we implement biological age. That's definitely very helpful for developing a healthcare plan if their biological age is high for their age, for their chronological age. They have the accelerated ageing showing that might mean that, hey, maybe we should screen more often.
Something could be happening so we can catch things early. And so this is where we come into that foundational baseline. If you're not measuring regularly, at least with like a wellness or something like that, you're not, you won't have that baseline.
You won't be sensitive to catching when there's changes and understanding what they are. And especially if there's lifestyle changes that, hey, is this what caused the health issue or not? It can also help with that.
So, this is where you'd want to bundle this into a routine wellness screening. And I would say the beauty of this is that we've started with mouth swabs, so it's not necessarily that it has to be tacked on that day in the clinic when they're already spending a lot of money getting their blood, their annual blood test done. Right now, we are working on blood tests that could be just in there with the screening, hopefully someday.
But for now, it's a great, something that they can add on when they're ready or at a different time, because money, you know, it gets expensive. So, in addition, the last one I would say is an objective measure of the benefit of prescription diets. So if someone has to go onto a prescription diet after they've been on that diet for a few months, it'd be a great time to check the DNA methylation, see what markers have changed because they've come on that diet, And is it giving a positive feedback that's doing what it should be doing?
Cause it can take a long time to observe those changes phenotypically. And so this is again, I'm gonna mention that baseline. It's best if you have a before and an after to look at.
So, if you can get them into a wellness routine that uses these markers, it's great. So, lots of research, going on around this biological age, and then, of course, diagnostics coming up. We're currently working with veterinarians, a lot of great companies, universities, and we're looking for more veterinarians to get involved and help recruit and participate in these studies.
We have a few grants that have been written. Once that money comes in, we're gonna need that to help recruit. So if you're interested in getting involved, please, please reach out.
We'd love to be in touch and work with you. And with that, thank you so much for listening. I hope you enjoyed the presentation.
