Behind the Breakthroughs: Rethinking Animal Research with the 3Rs

Elizabeth "Liz" Nunamaker:
The work the collaborative has been doing in the environmental health monitoring is actually having quite an impression on Charles River. Many people may not know it, but Charles River was actually instrumental in setting up the technology for the PCR based assays for establishing environmental health monitoring being a viable replacement.

Mary Parker (00:33):
I'm Mary Parker and welcome to Sounds of Science. Today we have an incredible conversation lined up with two passionate leaders making waves in science and animal welfare. Joining me are Elizabeth Nunamaker, Executive Director of Global Animal Welfare and Training at Charles River and Dr. Megan LaFollette, Executive Director of the 3Rs Collaborative, the 3Rs Collaborative, founded in 2016, is driving innovation and advancing animal welfare and research. Welcome Elizabeth and Megan.

Liz (01:43):
Thank you.

Mary (01:45):
Very happy to have both of you. the North American 3Rs Collaborative or the 3RsC was created to advance the education and science of the 3Rs, which is refining, reducing, and replacing the use of animals in research. So Megan, can you tell us about how this organization came to be?

Dr. Megan LaFollette (02:07):
Absolutely. So kind of this story of the 3Rs Collaborative began with some professionals. Actually it began at Charles River more specifically in that the leader of Charles River, Jim Foster wanted to do more for animal welfare and more for the 3Rs beyond just the internal Charles River environment. And so he kind of tasked this, he put this call out, and then a few individuals at Charles River actually got together. And slowly in that 2016 year actually started forming the nonprofit, forming a board, all of that. And then in 2017, we actually got our formal nonprofit profit designation. The point of it was to be completely separate from Charles River, even though of course Charles River remains a big member, an important part of the organization as a leader in the 3Rs. So that's kind of the short and brief, how we kind of got started initially.

Mary (03:51): 
Excellent. And just out of curiosity, how did you get started in this field? What's your background? How'd you get interested in this topic?

Megan (04:05): 
Great. But yeah, essentially I was getting a graduate degree. Ended up kind of switching to work with research animals. Got my Master's and PhD in laboratory animal welfare and behavior under Dr. Brianna Gaskell and Dr. Maggie O'Hare. And I've always loved science communication and teaching education in general. And although for some time I thought I would probably work in industry or just in kind of a research animal environment when I was graduating, there just wasn't good positions and locations that I wanted that were open. But there did happen to be a fully remote, actually this was pre covid, although I started when Covid started, it was fully remote, so I could live wherever I wanted. And it was a fellow program to work on this nonprofit that was actually perfect for my skillset and that it was all about human behavior change for animal welfare in the 3Rs, which is what I specialized in graduate school.

Mary (05:42): 
That's awesome. I'm sure we will get into how human behavior can affect animal welfare in just a little minute. But for now, I'm going to switch over to Liz Nunamaker. How do our ties Charles Rivers tie with the 3Rs organization help advance work with alternative methods?

Liz (05:59G): 
The 3Rs Collaborative is really this amazing opportunity for members from the Charles River family to get involved with the organization to share what we do in-house, but it's also an opportunity to interact with the other members of the initiatives to get their best practices in what they're doing elsewhere. So it's this great opportunity to exchange information, to share what we're doing to help improve the field as a whole, but also take information from other sites and improve what we're doing locally. It really goes along in demonstrating how sharing of information really makes us all better ultimately. And fortunately, we are able to have Charles River representation on the vast majority of the initiatives within the 3Rs collaborative.

Mary (06:51): 
And Liz, I know we've featured you many times on Eureka, and we're always happy to have your input. So can we just get also a thumbnail sketch of your background and some of your research interests that have led to this line of work?

Liz (07:04): 
So I took the scenic route to getting to where I'm currently, I am actually an engineer by training. I did my undergraduate and graduate education in engineering, and I was always really fascinated by the use of animals in research. And through doing my PhD, I was very interested in the ethics and welfare of the animals that we were using to support the research. And I decided to go to vet school specifically to become a lab animal vet so that I could focus in more on that. There was the development of the College of Animal Welfare, and I already knew that I was committed to lab animals. So I continued to work in my career to securing board certification by both the College of Laboratory Animal Medicine and the College of Animal Welfare.

(08:07): 
So having more of that, just having the background in both laboratory animal medicine and pure animal welfare and blending those together has allowed me to take what I know from these different fields and apply it. Lab animal medicine at its core is comparative medicine. What we learn in one species is always applicable in some way, shape or form to another species. And having the background in both lab animal medicine as well as animal welfare allows me to pull from all of these fields and apply it to lab animal medicine. So continually driving improvements, not that I'm necessarily the person doing it, but I know where to get the information to share it and encourage uptake of these different 3Rs initiatives.

Mary (09:04): 
Yeah, I would imagine that in lab animal welfare in particular, having flexible thinking is probably one of the most valuable assets you could have because our understanding of what they need and what's best for them is constantly evolving. And if we get too rigid about what we think is best, we're never going to be able to improve.

Liz (09:21): 
Absolutely. We're always learning new things that we can apply and make improvements. This is definitely not a stagnant field. If we get in the mindset of, oh, that's the way we've always done it, that's incredibly dangerous. Jim Foster actually recently said that during one of his recordings that the scariest words in the English language are, we've always done it that way. It's definitely holds true for lab animal medicine and caring for our research animals.

Mary (09:51): 
Love that. And speaking of improvements, Megan, what kind of exciting initiatives are you currently working on in the 3Rs Collaborative?

Megan (09:59): 
Yeah, so we do a lot. Right now. We've got, seven distinct topics that we work on. This includes kind a culture of care slash compassion fatigue, resiliency initiative. We just created and launched a 3Rs certificate course. That's really exciting and comprehensive for general training. On the 3Rs, we have an initiative on topics like micro physiological systems, environmental health monitoring, translational digital biomarkers, artificial intelligence for preclinical safety and risk assessment, non-human primate behavioral management. Those, I think that's all that we're doing right now. It is a lot. And then we do just general resources. We have a 3Rs sharing conference. Really our three main goals are just to make sure that the research animal community is supportive of the 3Rs and knowledgeable about the 3Rs and supported to implement them. Generally, we pick specific 3Rs topics that are high impact evidence-based and practical to really push and put a lot of dedicated effort behind individually. And then we also kind of just make sure people know about our organization so they can find our resources and get the help that they may need or want to make the important 3Rs Changes that impact the animals that we work with.

Mary (11:32): 
So you mentioned environmental health monitoring, so let's get into that. It plays a crucial role in fostering ethical and sustainable research practices, obviously. So how does the 3RC integrate environmental health monitoring into its broader mission?

Megan (11:51): 
So just for little background in case there's any listeners who are not aware, but so traditionally when we've got colonies of rodents, we have these cages of sentinel mice or rats that we transfer dirty batting into from LA colony cages. And then we test those animals, to then try to determine that colony health status. This makes sure that our research animals don't have any diseases that we don't know about. We obviously study a lot of diseases or pathogens on purpose, and we want to make sure that there aren't any ones that we are not interested in studied in our colonies. So this was kind of the traditional method.

Megan (13:23): 
So we want to make sure that our colonies are healthy and that we're studying what we mean to study. But now we have the technology through PCR to actually not have to use live soiled bedding, sentinel animals. And instead what we can do is if we have cages of animals, we can just swab either the cages or the racks directly, or we can still transfer that soil bedding to an empty cage. And in that way, just completely replace this other method. It's a really exciting and important and impactful 3R strategy because it's actually what we call a one-to-one replacement. So you just have to do one thing and it entirely replaces an animal method, and you just never have to use that animal again. Often replacements honestly aren't that simple. You might have to have three or four different methods

(14:29): 
Or more to replace a current animal model or method. So this is particularly exciting because you just need to do one thing very, very similarly often to what you were doing before, and you avoid that use of animals and institutions, depending on their size, they can replace hundreds, sometimes up to a thousand animal every single year that again, just will never have to be used again. So it's been really exciting to get to work with subject matter experts on this, institutions that work early adopters in switching and work with them to really champion their efforts as leaders in the 3Rs fields, and then provide resources and just sorts of different help and apply different strategies to get many, many other organizations across the globe to replace these animals in their research facilities.

Mary (15:34): 
So I'm curious from Liz, have you seen this change implemented already or have you seen other changes that have been suggested by the 3RC that you think have made a good impact on your work?

Liz (15:57): 
The work the collaborative has been doing in the environmental health monitoring is actually having quite an impression on Charles River. So many people may not know it, but Charles River was actually instrumental in setting up the technology for the PCR based assays for establishing environmental health monitoring even being a viable replacement. This work was done by Ken Henderson's group 10, 15 years ago, and it's been slowly building and building to what we know now as pathogen binder or more commonly the Shak ‘n bake method. It's a popular way to refer to it. Surprisingly at Charles River, we've only seen piecemeal uptake of environmental health monitoring. It hasn't been the default, but thanks to the work that the collaborative has been doing, more and more folks are recognizing Charles River as a center for excellence in this space.

(17:02): 
And we have it as an initiative for several of our business units to completely transition to environmental health monitoring this year, to eliminating all live animal sentinels that we might currently be using, which is really exciting. Other areas where it's been instrumental is in helping to apply pressure. This is completely separate from environmental health monitoring, but with refined handling, also known as low-stress handling, we were able as an organization to transition 87% of our vivariums over the course of 2024 with the goal of getting to a hundred percent transition this year. So I would say there's a lot of reciprocity, a lot of sharing of ideas that are helping to drive uptake of these 3Rs principles internally at Charles River as well.

Mary (18:00): 
So obviously, innovation and data-driven practices play a big role in this. So what kind of innovative technologies have been implemented or explored by the 3RC to enhance environmental health monitoring

Megan (18:17): 
There's a couple different ways to do environmental health monitoring. So we've come up with our own kind of vendor independent terminology for these. And there's two main ones. There's exhaust air dust testing or EAD, and you can only do this in Allentown or Tecniplast cages actually, because they filter at the rack level, which means that the dust from all the cages goes into the filter into this pipe essentially that leaves the rack. And so you just install a filter in there and then you pull it out and you send it to your diagnostic lab of choice and they do the PCR testing for you. So that's kind of one term. And then, yeah, what Liz was talking about, again, our vendor kind of independent terminology is sentinel free soiled bedding or SFSB. And that one you just kind of transfer the dirtying buttering, and then you either put filter paper or swabs or something to kind of collect those dust particles. Again, send it out for PCR testing, and then again, choose your vendor, send it out. They give you the results, they tell you what pathogens you might have, and then you're able to then either know what's in your colony, you can then work to address any positives and do the testing in that manner.

Mary (20:07): 
can you give me an example of how the data collected through this process through environmental health monitoring is used to inform decision-making and improve animal welfare? For example, have you come across a situation where you've seen a certain pathogen pop up and because you were able to identify it, you were able to isolate the source of it?

Megan (22:09): 
So see, Bovis is a pathogen that impacts nude mice, which are often used in cancer research.

(22:28): 
And if the mice have C.bovis, they may not be able to complete that study because it inhibits tumor growth, it impacts their skin, it can impact just how the mice are doing. And so it both impacts the animal welfare because if you have this pathogen as a nude mouse, you're pretty negatively effective. But then it also impacts the science because then you're not able to actually maybe complete that study. It's really counter to the 3Rs principle of reduction of using as few animals as appropriate for particular study. And so with environmental health monitoring, again, you're able to detect this and then implement certain protocols. And so the environmental health monitoring and the ability to do this pinpointing can really support all 3Rs from refinement to reduction to replacement, and just general scientific quality.

Mary (24:07): 
Excellent. Thank you. Thank you for that example. So it's right there in the name, 3Rs is collaborative. So let's talk about partnerships. How does the 3RC engage with researchers, institutions and any other stakeholder to align environmental health monitoring with the principles of the 3Rs?

Megan (24:25): 
Yeah, so we definitely do a lot of collaboration in our name. It something that we think is so important for actually just ensuring that change is really integrated throughout the whole field. So usually how that process starts is we start by collaborating with subject matter experts because we do really want to understand the 3Rs techniques that were forwarding.

(25:28): 
So we engage as staff with those subject matter experts, really understand what are we doing here? Why are we doing, does this replacement really work? How did the transition go at your facility? How did that work? What sort of barriers? What sort of costs, opportunities, benefits, etc, did you experience? So collaboration starts with those subject matter experts. Then we start to collaborate much more widely. So we have run a number of these really broad survey studies to get a pulse on what truly a wide range of people across the United States, Canada, the globe, are thinking about this technique. Do they think it's good? Do they think it's bad? Do you think it has evidence or not? Do they even know that it exists or how to do it? What are their concerns about it? So we understand that, and then we work together then with those subject matter experts to address those barriers head on, so we understand what's kind of standing in the way from this becoming more widespread, and what sort of resources, assistance, research, etc can we do to help it.

(26:44): 
So for example, for environmental health monitoring, initially there was a big concern, well, if I don't have Allentown or Tecniplast cages, I can't do this. It's not possible. Which initially there wasn't the Sentinel Free Soil Betting technique really well established. There'd been a couple papers on it. So we really encouraged more research on it. We then collected all that research. We educated people about the techniques. Then there was more research done, and then we were publishing more on it, and we got all the evidence together and publish a systematic literature review on the methods. And so to actually show hard evidence that this is actually better and more effective than the traditional Sentinel method. So things like that. Things like having SOPs developed, so then people know actually how to do it, how to make change at their institution. We highlight institutions that have changed so that way people really feel, oh, I don't have to be the first.

(28:02): 
We're like, no, there's all these institutions that have changed. If they can do it, you can do it too. And then also, we really like that too, because then we're able to showcase institutions that have changed and really lift them up as, Hey, you're leaders in the field. We're proud of you. We're thankful for the work that you're doing on the ground to implement the 3Rs.

Liz (28:53): 
And some other things that they've done too just to build on, because Megan's listed a lot of things. They started the 3Rs sharing conference to bring people together to talk specifically about the 3Rs. There's not a lot of opportunities to just be purely focused on the 3Rs like there is at that meeting. But then also setting up mentoring programs for low stress handling, refined handling. There's a number of members of that initiative who have quite a bit of experience, and they're getting matched up to individuals who are looking for help to making the transition at their site. And so again, forming those collaborations and mentoring relationships to help spread some of these 3Rs practices further.

Megan (29:37): 
And we also have a mentorship program for our environmental health monitoring initiative as well. And then we also, the other thing that I forgot is we also have general members of the 3Rs Collaborative, and we've got a membership membership program where organizations, whether they're industry or academic, can join us. They're able to get a few benefits from being members, but part of the big benefit of being a member of the 3Rs Collaborative is just kind of more intense collaboration with us, kind of having a direct line to your institution, understanding what's going on and kind of that back and forth. So we have members of specific initiatives of us generally. Yeah, we do a lot to try to make sure that we're not just taking one person's idea and running with it regardless of what everybody else is saying. We want to understand what's going on and how we can make change in a way that is positive for everybody.

Mary (30:39): 
So Liz, from the perspective of a member, what is Charles River's role in the 3Rs Collaborative?

Liz (30:47): 
So we wear a lot of hats in that space. As a large CRO I think it's very important that we contribute financially to the organization to ensure its long-term financial success, so that it's able to share these 3Rs practices. I'm fortunate to be a Charles River employee where the 3Rs are paramount. We are constantly making improvements in all three of these areas. And so as a member of the collaborative, I'm able to bring those experiences back to the collaborative and share them and contribute to the overall success of the field. I feel like it's a very symbiotic relationship between the two organizations, and it's more than just Charles River. I mean, we were involved in the starting of the organization and we continue to be a big supporter of the organization, but we need everyone to come and play in the sandbox, if you will. We need to have lots of different ideas and thoughts so that we can all work to improve the welfare of the animals we're working with. And moving the 3Rs forward as a field together,

Mary (32:04): 
What do either of you see as some of the biggest challenges for advancing environmental health monitoring in North America?

Liz (32:13): 
One thing I see as a challenge is we're not quite to the tipping point yet where everyone has made the transition. I'm always blown away by different locations telling me, oh, we've made the switch. We've made the switch. And so we are getting there. I think once we get past the tipping point and everyone is completely on board with accepting environmental health monitoring samples, I think that will go away, but it's going to take a little bit of time to get there.

Megan (33:10): 
I'd also add, so I kind of said, we've actually been doing, or we have done a three-year longitudinal survey to directly understand what the barriers are. We ask our survey participants what makes it difficult or impossible for them to actually apply environmental health monitoring in their institution. The good news is that the reasons in general are going down. There's more and more people who reply to that question with nothing. There are no barriers. We have already switched from year to year, so that's really, really positive. But there are still things that people do say sometimes there's people that are still not convinced of the evidence. They're really concerned that, oh, is this going to be as good scientifically as using sentinels or should I still use sentinels in my facility? I think with our systematic review, that is going to become difficult. That kind of reason is we've seen it decrease over time, and I think it's going to be more and more difficult for people to stand behind.

(34:31): 
What still sometimes comes up as reasons are cost. Although often institutions, if they do a full cost analysis, they often find that costs actually decrease when they switch But some institutions, if they don't use PCR testing if they're relying on older testing methods like serology or histopathology, those can be cheaper to do in-house. So that cost piece, I think always is a reality, although I really encourage institutions to really calculate, especially that personnel time that they're spending on a sentinel program.

(35:49): 
And then the other two main categories would be people in just that it often, you can't usually make decisions unilaterally in a research environment. There's a lot of people and a lot of different hoops and people that you need to convince. So maybe even if you personally are on board, you have like five, however many other people that you need to convince that you need to train, that you need to make this conversion. And then that leads to the smallest barrier, which is just time in that often we're super busy and unfortunately that we've always done it this way. It takes no time to continue to always do it this way. Technically, you're like, well, I don't have to train anything new. Whereas it does take a little bit of time to figure out, okay, well, we're going to change this method. What vendor are we going to use?

(36:49): 
Which method are we going to use? Okay, I need to retrain all these people that I were trained in this other method and just have some systems. And so the reality of making change is it does take a little extra time, and I think it a hundred percent should be prioritized. But sometimes if you're in the midst of staff shortages or various crises or big times, it can be a little bit challenging just to find the time, which is again, where our organization really comes in, is we've done all that I think we possibly can do to try to make that switch as quick and easy and the least amount of time that you can.

Mary (37:33): 
So switching gears entirely, let's talk about digital biomarkers first. For my benefit, what are they?

Liz (38:09): 
so digital biomarkers are their own thing, but a way to get people thinking about digital biomarkers is by referring to Fitbits and Apple watches. Almost everyone in the world is wearing a Fitbit or an Apple watch, or it feels like it. So people wear Fitbits and Apple watches to get additional information about themselves, how active they are, maybe their sleep quality, maybe their body temperature. And in the elderly community, oftentimes they wear them for the heart health capabilities because it's really good at picking up arrhythmias. This is something that's part of their environment. They're wearing it on their body all the time. They don't really notice it. It doesn't change their behaviors. But when you start looking at the trends in that data over time, either yourself or in conjunction with your doctor, you can start to get a lot of information about your basic health, whether that be you're very healthy, maybe you're spiking a fever and you're going to get sick. Maybe you have some sort of heart arrhythmia or some sort of cardiac disease that's progressing. So these devices are essentially a digital biomarker of your day-to-day life.

(39:27): 
Yes, we're not putting Fitbits, and Apple watches on our mice and rats, but we are able to have smart technology integrated into their housing that gives us information about how much they're moving around their cage, what's their respiratory rate look like, what's their body weight look like. It gives us a lot of information about just basic health of the animals. And by understanding that in animals that aren't manipulated in any way, at least externally by someone handling them, we can pick up on very subtle changes. So we can identify a disease progress, a disease progressing in those animals. Maybe we can pick up on early toxicity. We're able to identify these biomarkers, if you will, of disease processes or toxicity reactions that we wouldn't be able to pick up on otherwise. And it's just by simply collecting this information about the animal in their home environment, in their undisturbed home environment.

Mary (40:32): 
Yeah. So Megan, back to you. How do digital biomarkers contribute to the 3Rs's principles, and what kind of advantages do they offer in bridging the gap between preclinical research and clinical outcomes?

Megan (40:44): 
So digital biomarkers are really exciting. We actually really emphasize and have an initiative focused on what we term translational digital biomarkers in that we really define these digital biomarkers as it's data collected continuously from unrestrained and uninstrumented animals in their home cage environment. So that really contributes to refinement. And then the translational piece is that this should be an objective, quantifiable measure of either physiology or behavior that relates to disease progression or therapeutic intervention. And again, this ideally is clinically relevant and actually translatable between preclinical studies and the clinic. So this contributes really directly to reduction.

(42:03): 
This idea that we should, across the entire research environments, all the studies be using as few animals as possible, but making sure that the animals we do use, we're getting good data from that is actually translatable. And so ultimately then what these digital biomarkers can do is that maybe they're looking at, I think a really good example is, yeah, we take our temperatures when we as humans get sick, but a lot of times we realize we're getting sick because we don't feel well. And that's a lot of times in a lot of diseases that we're trying to treat. What we're trying to treat is we're supposed to make people feel better, not have X heart rate, X temperature, whatever we typically might measure.

(43:13): 
And typically that feeling well is related to our general activity. When we feel good, we're moving around our house, fine, we're going on our walks, we're exercising, et cetera. When we don't feel good, we're sitting on the couch, we're stuck in bed, we're sleeping a lot, et cetera. So one where way there's digital biomarkers can really help by getting an idea of that activity from an individual animal or just over time you kind of get a baseline, then you can see a change. And a lot of times in these studies, you can actually detect that behavioral change of decreased general activity sooner than you see those more traditional measurements like say, temperature or weight loss is one that we look at. And that way you can potentially stop a study sooner before the animals have to go into more stressful phenotypes, i.e., refinement, which is great.

(44:14): 
But then also, again, maybe you might not see some of that weight loss or something. Maybe your intervention isn't seeing that result, but you're seeing that change in activity. So again, maybe you're having a translation or an effect on clinical patients really translating to translation. And then the other way that you can use these, that's both very much related to that, but separate. But as the research animal staff who's just one of our jobs is to make sure that our research animals are healthy in general. And that again, that we're intervening at an appropriate time in a study that everything is good, that everything is healthy, etc. And so you can sometimes use these digital biomarkers to just understand your general colony health and which animals maybe are going downhill faster than others. And so you need to have that intervention so you can really enhance their welfare in that way. And kind of that, I think the behavioral piece is really interesting because it can be challenging and very time consuming to study animal behavior. If you're just watching videos and manually coding something, it takes a long time and a lot of manpower. But if you're able to use these digital biomarkers, you can maybe run studies faster and with fewer staff or just able to process things and see things that we weren't maybe able to see before.

Mary (45:50): 
My next question gets into some of the cutting edge technologies that are used for these.

Mary (46:36): 
Are there any other cutting edge technologies that kind of fit this bill?

Megan (46:41): 
Yeah, I mean, so those digital monitoring technologies, they really vary in how they might apply it. Some of them are using kind of video and then sometimes using machine learning or deep learning algorithms to process that data. Some are using RFID chips where those are implanted in animals and maybe they're just collecting temperature remotely, or they're looking at, they use that RFID chip in combination with kind of a plate, or sometimes they're able to have something on the bottom of the cage that isn't even connected to an RFID chip, but they're able to just see the activity piece through. Through different ways, they're actually able to record that data just from the bottom of the cage and not use video, which is sometimes actually nice because that video processing and storing a video takes a lot of space, adds that complication.

(47:40): 
So sometimes this is kind of simpler can be more cost effective. It's just different applications. So some are video, some are more hardware based, some are more physiology based. Some they have these cages where there's operant studies where maybe they're training the animals or they're having the animals do some sort of task. But there's some automatic sensors that basically do the training themselves and record all that data and see how the animals are interacting with different pieces of the caging. There's lots of different kind of creative technologies. Basically, if you want to measure it, there's different ways to measure it. And whether it's sound pressure, video, heat, infrared, there's a bunch of different technologies that are all, again, then used to understand whether it's a cancer model, an aging model, an infectious disease model, whatever, that then we're really able to understand that disease progression often in just a quicker way and a deeper understanding than maybe traditional measurements.

Mary (49:08): 
I love that. Liz, anything to add on this topic?

Liz (49:12): 
I mean, every model has its unique needs. And so a lot of these systems, it's a blend of commercially available and homegrown sort of experimental systems, but it's modular. So you can add back what pieces you need that are relevant to the animals that you're working with, to the model that you're working with. So you can be collecting those digital biomarkers that are truly relevant to the model that you're working with so that you're not just collecting, because the data becomes a whole headache in data management and storage. And so making sure that you right size what data you're collecting is really important so that you can ask and ask informed questions and get better data.

Mary (50:01): 
Yeah, good data is definitely the key for any of these projects to succeed. I mean, it's kind of the backbone of well-done machine learning is you have to put in good data before you're going to get a good result.

Liz (50:13): 
Exactly. For sure.

Mary (50:15): 
So can you share an example of a successful collaboration or case study where the implementation of digital biomarkers has improved the 3Rs goals?

Megan (50:27): 
Yeah, so I would say that, I mean, there's many, many examples. I think I could pick a lot of different ones depending on the particular technology, the particular animal research. I'm just going to pick one from. We have a paper about the emerging role of translational digital biomarkers in the field. And so if anybody listening wants to learn a little bit more about these technologies, dive into a little bit deeper, you can do that. But one of the things that I'll mention was there's an example of using a home cage monitoring system with an acute colitis model. So this is a GI track model that's not exactly super comfortable for the mice. So we really want to detect any changes earlier. And so they use this home cage monitoring system. They didn't have to do anything directly. They didn't have to remove the animals from the cage.

(51:32): 
It was objective, it was continuous, it was non-invasive. It was compared with kind of gold standard parameters and detected earlier. And this is able to both improve the animal welfare. So they're able to actually stop that study and intervene with rescuing the model earlier, but then also enhance scientific quality because you're not having to kind of do that intervention or pick up the mice or add this additional stressor to the animals. So that's just one of many. There's other examples in that paper, and we see more and more examples of this being used over time.

Mary (52:15): 
So what do you both envision as the future of translational digital biomarkers and how the 3Rs Collaborative is going to stay at the forefront of this?

Liz (52:26): 
I personally would love to see the technology get cheaper so that we can integrate it as a base model into all caging options that we have so that we can get to a point where it's like, okay, we're using this model. What data do we want to collect? Oh, boop, boop, boop, and we picked those three things that we want to monitor. And it's just there. It becomes the standard of care. I do think it's doable, but it's going to take a while. The smart caging options have been available for a while now, but they've just been so expensive. It's been a bit cost-prohibitive, but it is an area where a lot of really smart people are putting some concerted efforts into identifying better ways to do the data management piece, as well as provide affordable options to integrate into the home cage monitoring. So I think it's on the horizon. It's going to take a little bit longer, but I would love to see us get to that point where we're not having to manually take the animals out of the cage every day to do routine monitoring, being able to get more basic information about the animals in their home cage. I think it's going to be much, much better for animal welfare and improve the quality of the science coming out,

Megan (53:45): 
Coming to build on kind of what Liz said is, so at the 3Rs collaborative, I think we're going to continue our work with our translational digital biomarkers initiative. One of the things that we have done recently as we've focused on validation, verification of these technologies, looking towards that, both just general confidence in them, but ultimately regulatory acceptance of the measures. So I think in the short term, if we could have a future where there's some guidance from the FDA on this, or there's some positive examples of these digital measures, digital biomarkers actually being used in, say, an IND application or used more concretely in drug development that would help encourage more institutions to invest in them. And so we have this paper on that process that I think might help with that and might just build the confidence in the data, which is really important piece of it.

(54:56): 
The other thing in the short term that we're working on is establishing that return on investment or thinking more about that because as Liz said, what has stopped some institutions from engaging is that cost factor. And so if we can show more examples where, no, there really was a positive return on investment from these technologies, we're going to see more and more engagement. And I think in the short term, a goal that I think I would love to see is just each, every organization having a small number of cages with some of these technologies so that they can try it out so that they can see for this study or for this application, it was really beneficial. I always encourage when you're making change to start small, to figure out what are the unique needs of your institutions. And this is a way too, this is what I've heard at some other institutions too, that they may not have these throughout their vivarium yet, but they have cages that then they book out for these studies because they have found them really valuable for a particular application, a particular scientific question. And I think even having that to start would be a really positive thing for the field to just utilize these more, get more publications on them, and demonstrate their value for translation, for reduction, for refinement, for scientific quality, animal welfare, really all of it.

Mary (56:39): 
Well, Liz and Megan, thank you so much for being part of Sounds of Science and for your insight on this topic. It's been a real pleasure talking to you.

Liz (56:47): 
Thank you. Thank you so much.

Mary (56:49): 
Elizabeth Nunamaker, executive Director, global Animal Welfare and Training at Charles River and Dr. Megan LaFollette, executive director for the 3Rs Collaborative. Stay tuned for the next episode of Sounds of Science. Until then, you can subscribe to Sounds of Science on Apple Podcasts, Spotify, Stitcher, or wherever you get your podcasts. Thanks for listening.