Good afternoon everyone and welcome to today's webinar kindly sponsored by My RNA Diagnostics. Our topic today is the use of microRNAs as biomarkers in heart disease, and I have the pleasure of introducing you to today's speaker, Doctor Eve Hanks. Eve is a.

Surgeon and research scientist. Her key skills and training include PhD and postgraduate certificate degrees. She's an innovator, entrepreneur, and entrepreneur, and she used her clinical background for conception, development and implementation of this new testing technology.

If you have any questions or comments for Eve as we go along, please hover your mouse on the screen, click on the Q&A box, and we will cover as many of those as we can at the end of the session. But without further delay, let me extend a warm welcome to the webinar, Betty, and over to you. Thanks, thanks very much, Jackie, and, good afternoon.

Welcome to a session, a very scientific session actually for a bit of a change, on microRNAs. And I'm going to take about 45 minutes to just give you an overview of microRNAs as biomarkers and as diagnostics. We'll have a little look and put some clarity into this.

Because I think if you've been around at all in the last, 10 years, you'll have heard about microRNAs. So put a bit of clarity and, and some examples in place. And, I think that we'll probably have about 40, 45 minutes, so plenty of time to have some questions afterwards.

So as Jackie says, if you'd like to enter any questions, into the chat box, and I'll try to get to those as well. So myRA is a new company. We are, out we've spun out of Scotland's rural college, SRUC.

Now, if you're not from Scotland, that SRUC are a diagnostic testing, veterinary diagnostic testing company who work with the Scottish government alongside APHA. And that is where I was working when I came up with this idea, for the new diagnostic test that we've developed. Prior to that, and while I was also working at SRUC, I was still a veterinary surgeon in practise, and I did my undergraduate at the University of Edinburgh, and then in Liverpool I did my postgraduate certificate and finally my PhD in infectious disease in Glasgow.

So I'm a research scientist, a vet, and I have a clinical pathology background, and that's really these threads came together, as Jackie was saying there, and I, I suppose the right place at the right time, and I managed to have a new look at how we, deal with diagnostics in the veterinary fields. So, microRNAs, you may have heard a little bit about, perhaps liquid biopsy is a, a term that goes around just now, and the reason for that is microRNAs have a big role in cancer, so we'll have a look into that. They're really very important.

So, I'm gonna tell you why we should be paying so much attention to microRNAs and why as clinicians, as veterinary professionals, and nurses, we should really be looking into this area as new developments emerge in human and veterinary fields. And I think I'm going to predict a little bit of our, our new likely developments in the future as well. So microRNAs are interesting.

The basics are that they regulate a variety of functions. They're very small, they're non-coding pieces of RNA. And we can find them within cells and also they are cell-free, which means they're circulating in our system.

We know what they do in the sense that we understand that they are important for growth and development. They manage stem cells, they tell cells when to die, apoptosis, and they tell tissues what they are to be. But we also don't know what they do, in the sense that there are a lot of them doing a lot of things that we haven't understood as yet.

So generally they are thought at this time to regulate all of our cell processes. They're considered to be key regulators of immunity as well, which is important for us as clinicians. And it's been nearly 30 years now since they were, believe it or not, 1993 is nearly 30 years ago.

So they were discovered initially in C. Elegans, and C elegans is a free-living nematode, a cousin of Heunchus contortus, one of our, our sheep nematodes, or ruminant nematodes. And when they were first discovered, after a period of time, there was an understanding that, these molecules were redefining what we thought genes did and how they were regulated.

So they act on messenger RNA and what that means is that our DNA will trans transcribe messenger RNA which results in protein, and they don't act on the genes, they act on that intermediary stage, the messenger RNA predominantly. So this gives us a really quick way of responding to stress. Genes can be a bit clunky to turn on and off, but if you're looking at messenger RNA that you can have almost instant, results here.

And they're important to us, really and. All of our life stages, they are found in embryos right through, until death as well. So they are very, they're very active.

There is a lot of work, looking into my arms and microarneys just now. They're little hairpin structures, as well as what they look like, and we'll have a look at that in more detail in a second. And of course, since they were first identified, there has been, myriad research done in this area, and, of course, weed to the human field, but there's also been veterinary research as well.

It might surprise you to know that on Mirbase, which is the online database where you can record a new microRNA or look into existing ones, that there are over actually almost 40,000 entries, currently logged. So there are a lot of microRNAs, and partly that's because they are in all species. So they are in, cows and sheep and dogs, all mammals, humans, they're in viruses, plants.

So we can find these little molecules everywhere. So to help a little bit, there's now some standard nomenclature so that we can all keep track in the research community of who has found what in what species. So, for, I suppose, initially, it was thought that microRNAs were made in the nucleus and remained within the cell.

And they undergo some extensive processing actually to make them or to take them from that pre-microRNA stage to a mature microRNA. And then interestingly, for me, particularly as a diagnostic you know, scientist, what we then found is that microns are actively exported out of a cell into the bloodstream, urine, milk, saliva as exosomes and vesicles. We have literally no idea why.

It's assumed there's perhaps some sort of signalling in between tissues, but other than that, it is still up for debate why we spend time and effort putting these microRNAs outside of cells. So research at the moment is intensive, and it's probably one of the largest research areas that's ongoing, is microRNAs. So there's constant identification of, of new microRNAs and but there's clearly lots yet to know.

When you do discover a new microRNA, if anybody is thinking about getting into this area, then unfortunately you don't get to name it as you do in some areas of science. In fact, it has to just be sequentially numbered. So you may see a notation such as this HSA standing for Homo sapiens, and then MI R to tell you that that is a mature microRNA, not a pre-microRNA, which is important.

And then whichever number it's been given. Now, the reason that that's important is, that mature RNAs can come from different, from different mature RNAs can come from the same pre-microRNA. So the processing there that is done to the pre-microRNA actually determines which mature microRNA you get from there, so it makes it a little bit more complex.

But interestingly, they are highly, highly conserved. So what that means is that the micronAs we see in humans, we will see in dogs, we will see in in other mammals as well. So that helps us in terms of research and development, which is really important.

And 30 years ago this all started and clearly it was not as easy then as it is now, and even during my PhD this was a more difficult process. Initially research was hampered, I suppose, because to collect microRNAs, you initially had to have a dead subject. So you were postmortem tissue samples most of the time.

It was extremely easy to contaminate what you were doing. So genetics is a bit less robust than protein research and from the point of view that it's very easy to introduce some alien, DNA or RNA to what you're doing. And you didn't know if it was contaminated because typically, you had to wait about 6 months for a specialist lab in North America to come back to you to tell you your results.

So you can imagine if it was contaminated that that really puts your project back. And the samples are also difficult to collect, difficult to store, and super fragile. So whenever you have RNA, you have RNAs, your enzyme ready to take those down, and that's true also in tissue and you couldn't just freeze this to stop that process because you damaged your very fragile microRNAs.

There are some significant challenges at that point for the industry. But everyone persisted because we know that microRNAs are actually not a nice to have, they are essential for life. So when you make knockouts, and when you've taken your microRNAs away from the animal, and then they die.

They effectively can't live without microRNAs. So they are essential, therefore, it's essential for us as a research community to understand the fundamentals behind them. And with so much that they regulate, we're looking here at microRNAs having a driving force behind stem cell differentiation.

They can promote growth, they can inhibit growth, they can ask cells to die, to proliferate, they can create blood, they create organs. So when we have this type of, role, we really need to understand them both in health, but also, of course, in disease. So we'll have a look at a couple of simple Examples first of all, because sometimes in science it's nice that we can find just a simple pattern.

So for example, microRNA one, so ME one, actually, when you knock that out in mice, we can see that MER one is essential to heart health. So if you don't have that, then the heart is defective. So that's a lovely straightforward correlation, causation between one microRNA and one organ.

And we can see that repeated in different species and, and in different disease types or different organ types also. So for example, MR 203 is essential for skin differentiation. It's a bit like at school when we're taught biology, when you have one gene for one eye colour, so it's important.

We also know that sometimes they're driving other things within the body and they again, are, are essential. So my 127 really does have to be around for lung inflammation, something that is being heavily investigated currently with COVID, and also, MER 375 is quite central for insulin secretion, another big public health issue. So we do have these straightforward relationships between one microRNA and one process, which is nice.

And when we look at this and, and we consider that all disease processes are regulated by microRNAs, you know, this then means that we're going to have to have a better look at some specific diseases, and most of these unfortunately don't have just as simple a correlation as one microRNA to one disease. Now, by far the largest area of research, as we've said, is oncology. So, tumours, cancer, neoplasia, it's correctly called, of course, has been studied extensively, because microRNAs are so influential in growth and depth of tissue.

And then we'll have a little look at some infectious disease in the human field, some non-infectious disease, because that's where predominantly research has been ongoing, of course, is the human field, and but it's not exclusively there and we do have some veterinary papers to look at also. So cancer is not simple, and it's multifactorial, of course, there are a lot of things that can lead to a patient developing cancer. But we know and understand now that microRNAs play an increasingly recognised role.

So we know that some are known to, to repress tumours. For example, it's a really big finding through the use of, of apoptosis pathways and through influencing proliferation pathways, they do the opposite. Actually, of all the microRNAs we know at 50% are involved in oncology, so really that's where studies are targeting just now.

So there are I suppose a plethora, you can really fall into a well here if you would like to know more. And some cancers we now know as well can't occur without certain microRNAs which for the future, we'll talk about the end of this presentation. It's just absolutely fascinating to think if my 155 isn't there, the patient cannot develop lymphoma.

So they really are instrumental. And this is just a a jump to the side slightly where there is a blood test being rolled out in NHS England using a similar technology, where they're actually looking for rather than self-free RNA, they're looking for self-free DNA. So there is a slight difference, but this is still genetic testing and The results are really promising, and there's a couple of things here that are interesting.

They can detect disease early using this, and this one blood test can detect 50 types of cancer. They're not detecting it that often, in fairness, 51% of cases, but that will improve as the technology improves. And I thought I thought was interesting is that the only wrong way detect, or false positives are extremely low, and if you look at how specific Those cancer figures are that it actually, looks very promising indeed.

So who knows, maybe you or someone you know may be involved in the pilot, which is not exactly what we're looking at today, but it's within the realm of the genetic testing that I'm talking about. So, We now understand that we have got, I suppose a significant input, between host and a pathogen microRNAs when a disease, an infectious disease occurs. We understand, of course, a little bit about, our innate and adaptive immune responses and how they talk to the pathogen talks, to the whole and and the other way around.

And we now understand that actually a lot of that communication is at the microRNA level. MicroRNAs control or finely tune every aspect of our immune response. It's regulated by microRNAs.

So I think initially I thought this was a fine-tuning, but actually, they're more like project managers, if you imagine them like that when it comes to infectious disease, because they have so many multiple interactions with, so many different parts of the immune system, every inflammatory pathway every Cell, and if we chemokine. So really, when you add them then the pathogen interactions that are also going on, the viral micronies that are talking to our micronies and changing them, there is a really complex picture that appears here. And I think that that idea of viral mRNAs, we, we know that viruses were able to control our response to them, but we didn't necessarily understand why.

So let's have a look at a couple of human stories just to a couple of human diseases just to give you a bit of background. So Epstein-Barr virus is a member of the herpes virus, herpes virus 4 in humans. It's a global disease and it can cause just mild disease, but it can also cause months of fatigue, similar to several viruses, including coronavirus.

And one of the things that it can do is the viruses can actually hijack our microRNAs and control our own cells in order to allow it to remain latent, which is fascinating, and then it can reactivate, we're not sure why. There are other things that the virus can, can cause a viral infections such as this can cause, such as Ian-Barre syndrome, which you may have heard of, and that's when you're, it's an autoimmune condition where you start to attack your own nerve cells. So again, we think there's quite a lot of research, but the community think that this may be a derangement of microRNAs in particular.

There's no treatment, there's no vaccination, and so, therefore, would it be something that we can, if we can understand a bit more, might open up new treatment pathways. And then another example, very well-known example of course is HIV. And we know that HIV, I suppose enjoys, enjoys interaction with our immune system in particular.

We know how much they use, can understand now how much they are using microRNAs to allow them to allow itself to replicate. The viral microRNAs in this case actually act against our genes. They do a bit of hijacking of our own, but they also have viral mRNAs acting against our genes.

And there's 3 stages to HIV. So we have an acute stage, then chronic, which is the kind of remission stage, which is where treatment is targeted and and people can stay in that now for life, and then the final stage of AIDS. Interestingly, when we think about microRNAs and as potential disease targets, then it's, I always think it's good to think about, well, would they be as useful as we think.

And in the USA just now, every 10 minutes someone is diagnosed with HIV. I didn't, I suppose, fully appreciate the problems that are still out there. And 1 in 6 people with HIV it's projected, don't know they have it.

So even with good diagnostics, we're not picking up everyone that is carrying disease and therefore we can't treat it, so we're starting to, to think about, well, in the later stages, is that when we need to have some other arsenal in order to treat or even eradicate and cure HIV and could that be involving microRNAs. So we'll come back to that later. Then another infectious disease, I in no way mentioned Geronimo, but we know that tuberculosis is difficult to detect, and I'm sure that you are aware that TB is a very successful pathogen.

It's actually very difficult to detect, in particular in children. So in the human field, we miss 30% of cases because of problems with diagnostics. And incredibly 25% of the world's population has TB.

So it's the top 10, 1 of the top 10 causes of death, and kills more people than HIV and it's preventable, you know, this is the thing that's difficult is that it's so preventable if we had the right infrastructure. There's also a big public health concern with multi-drug resistant TB, so a real crisis there and microRNAs are known to be able to inhibit bacterial growth of TB. So there are microRNAs that can slow down and possibly even reverse disease, working alongside or instead of antibiotics.

That's a really interesting area. And you may have heard that Geronimo's postmortem is negative for TB at present. So we do have problems in the animal field as well with, with reliable diagnostics, as you will be aware of.

So, then if we move on to some of the non-infectious diseases, and again, we're just, I'm giving you some background here to know a little bit more about the potential of microRNAs because you're going to come up against them in the future. So Let's have a look at some of the processes where microRNAs are important. We've got autoimmune disease, genetic diseases, metabolic diseases, all of which are causing some problems, for example, multiple sclerosis.

Interestingly, this is a cold climate disease, so being from Scotland, it's something that definitely occurs more often in Scotland than it does down near the equator. And the reason for that is that, just out of interest. For every 10 degrees north you move from the equator, your symptoms of MS will be onset 10 months earlier.

So for every 10 degrees you move north, you're 10 months earlier with MS symptoms. We don't really understand why. But I think more importantly for this particular discussion is that in MS patients, 42% of our patients with MS are initially misdiagnosed.

So this is another disease that would benefit from treatment in the initial stages, but is not often picked up. It's therefore hard to monitor and hard to distinguish between the different types of multiple sclerosis that occur. So, microRNAs such as MR 155 may well be positioned correctly to help to understand progression, relapses.

So really interesting, area for microRNA research. And then, of course, type 2 diabetes is a massive public health concern across many countries. There's around 400 million adults living with diabetes, and so, again, an area that we really would like to have better treatment for.

And interestingly now, just in terms of public health, the way that our country can suddenly be burdened with a problem such as this is that China is now the country with the highest incidence of diabetes. They have 1 in 10 adults suffering from diabetes, whereas 10 years ago it was almost unheard of. So it's really important that, yes, of course, diet, exercise, there's genetic factors, but it really is important that we try, as a research community to, to come up with new treatments, and of course, microAs are involved again in this.

So it gives us lots of, of food for thought. So that's very interesting, but we are predominantly veterinary professionals, on this call, so, you're starting to see a pattern I'm sure, in terms of what could be done with microRNAs and hopefully a little bit inspired to think about, working in research in this area. But, in terms of what we're doing, let's look at small animals first of all, what are we doing?

Well, not enough. There's so much research that could be done. And when we are looking at at disease, and microRNA patterns, we're finding some really interesting results.

So when you think about the challenges of diagnosing a cancer, we're talking about biopsies, looking at slides under microscope, and asking specialists to do these types of jobs, we can improve that in a lot of cases. And canine lymphoma, B cell lymphoma, a study recently had a look, at, a diffuse large B cell lymphoma. And found that using microRNA 3 cell-bound microns, so this is not blood samples, they were able to 100% differentiate between healthy patients and those with lymphoma, 100%.

So I mean that's just unbelievable for the sort of size of studies that they will have been funded to do, you know, we're not talking about tens of thousands of patients, it's incredible results. And then we can look at mammary cancers and intestinal cancers, for example, in dogs as well. T cell intestinal cancer would be another one where it is possible using only a few, a handful of microRNAs to differentiate between healthy patients, intestinal tissue, intestinal lymphoma.

And enteritis patients. So you can actually even if you add an inflammatory component to this, you're still able to use my carnies to tell you what's going on. Now, of course, it doesn't happen in every type of cancer and that's why I put oral melanoma there, relatively common, of course.

And, we unfortunately, so far in the research that's been done, it's not as clear cut. So there is a, a differential expression of some microRNAs and melanoma patients, but it's not possible to assign them to a healthy or non-healthy group in the same way it has been with lymphoma. So there's lots and lots of work, to do in this area.

And then of course not forgetting production animals, so I, I'm very much a mixed vet, so I've worked with a variety of species and some of these concerns that we have are some of these diseases that we're targeting or reduction of waste and greenhouse gases and, and helping the economy, and, you know, microRNAs fit into that little bubble as well. So, for example, hereditary muscling and sheep. It does appear that that can be controlled by microRNAs, whether it should be or not, that's a different discussion.

And then disease control on large units, I just randomly picked a, a pine virus, but of course, there are, lots of, of production units where, the use of perhaps antibiotics should be reduced, the use of firefighting veterinary medicine and to improve the economics and, and production. So microRNAs are also being looked at in that field, but I'm not going to, to diverge too much into that for today. So I think we can agree that they're worth investigating, but until now, it hasn't been actually very easy to do that because, as we said, we need a postmortem animal, which is not ideal for diagnostics.

However, then we realised that they were in biofluids. Now microRNAs are in all biofluids that have been looked at, and they're in urine, they're in milk, they're in faeces actually, they're in blood, saliva. And when they are found in biofluids such as serum, they're highly stable in comparison to tissues.

You can freeze and thaw them. There's not much in the way of RNAs ready to break them down. And so what you can actually do is take a sample from a patient, store it at room temperature, and send it to a lab.

And so that really for me was a breakthrough because we knew we wanted to use microRNAs for diagnostics and this was the first time we've been able to do it. However, what we need to do is test it in a timely manner as well. You can't wait 6 months for results as a clinician.

So, unless it's a ringworm, panel. So what we then did was had a look at SRUC and with myRA we had a look to see what we could find in terms of microRNA assAs that could be run in-house in Scotland. And Aamer North American company who make a bespoke panel of microRNAs.

So what happens here is you tell Adcam that you would like X number of microRNAs and for us we have about 23 or 24 microRNAs per well, and then you add in your serum sample, and these microRNA binders will capture the microRNAs you're interested in. And then it runs really very much like any PCR or Eliza might do. So we have a PCR stage.

We fluorescently tag the microarnas and that are left after the washing process has been finished, and then you're able to quantify what micronnes are in that sample. So we can now create panels, we can validate the panels, we can, I suppose, run this protocol in 7 hours as opposed to 6 or 7 months, and that suddenly makes this whole thing viable. However, initially, it didn't work that well.

And the reason for that is that it's just the data that you get is just too complex. So initially we had differential expression of 23 different microRNAs and which were all differentially expressing from each other within a patient and then from the patient next to them. And so PCA plots are just not going to cut it when you're looking at this.

And when we introduced, we had a statistician who introduced an artificial intelligence. Mo, it was a literal bingo moment. We were able to then see that the patterns we thought we could maybe pick up manually could just be created within 10 minutes in front of our eyes.

And so we have been able to develop a heart test for both dogs and cats using this new microRNA technology, which is astounding. So we're extremely pleased with this indeed, and it's highly accurate. And of course, therefore, we're bringing it to market.

So that's one of the reasons I'm telling you about it, as well as trying to get more vets into this area of research. We're now selling the first microRNA test of any company anywhere in the world, either human or veterinary medicine. So that's a fantastic achievement for us here in the UK and in Scotland's Science Park.

And what we're doing is this commercial test that you have to register with us and we're giving people support because it's a new type of test. So it's free to register, but we do ask you to do that through the app. And then Nemo, which is our, our brand name for this test, or Nemo heart test is the first of our offerings.

That now we know now what microRNAs do and what we're doing here is detecting early stage heart disease using these serum biomarkers, microAs, and they undergo AI modelling. And it's relatively simple. We just ask you to submit serum samples, to us, but if you don't mind pre-separating that serum before you send it, that would be fab and next day post for sure.

And then you get your results back through the app. Now, we're going to be working on this, updating disease. You may see we've got a cycle here where we're asking you to receive your results and update your diagnostics.

And the reason for that actually, I'll come back to in a second, but the reason for that is because we can do more with this test than just detect disease. So, I'm obviously not doing this alone and I just a huge shout out to the team, not all of them are shown here, but the, University of Liverpool has been absolutely instrumental in helping us in the cardiology side, to, to get our samples in and. Them to have our our test validated and Hay here is head of the AI side.

He has, we're very proud of him. He's been doing a lot of work for the Scottish Government with COVID and then also working with us, a very distinguished member of the team. So what we can do with microRNA testing at the moment, this is when it does work really well, because it picks up early disease, we are, suggesting that you would want to use this for predisposed breed screening.

You would want to use it for an assessment of heart murmurs, when you first pick them up with no clinical signs. You may want to run wellness checks on occasion on, on certain, dogs regardless of species and cats as well. I think pre-op checks, and it does take a day to day or two to come back to you, so you would have to plan ahead.

This isn't your standard, you know, make sure the kidneys OK. But certainly if you did have suspicion of heart disease and that was one of the reasons you were using an anaesthetic or you're perhaps going through quite a long process, quite a long procedure, orthopaedics, for example, then this fits nicely there just to make sure that you're got a healthy patient before you start. Cats with no murmur, of course, cats don't often tell us that they've got heart disease, so it's useful if you're suspicious, but you don't have any proof.

And it's so much more accurate in early stages than pro BMP plus because it's AI, it's really, really easy to interpret. We tell you whether or not your patient is diseased, which is really useful. And there are, I suppose that there's sort of that grey area, you're slightly above the cutoff or you're in the cutoff.

And so nice easy user interface as well. What we can't do yet, but we would like to do, and we would like your help, if you don't mind to get our, our AI system to that point. And I think this will be common with microRNA testing and from various companies as other people bring these out, is that we are not yet able to categorise disease, but we're very close.

So this, because micron. Vary within tissues so specifically, but then they vary within tissues within organs as well. We're actually able to see not just yes, your dog has heart disease, but we can say yes, your dog has mitral valve disease, it has dilated cardiomyopathy, it has aortic stenosis, but we're only about 75% accurate with that and cats, as well, we can do the same.

So what we would like you to do when you come back onto the app to submit another sample is if you can tell us what the dog or cat was diagnosed with just a line, just a couple of of words, then we can tell AI and that will help update and then we can give you a test that's even better than it currently is. So there's a little bit of community spirit there and it's completely up to you. We'll give you some money off if you do it, and if you don't do it, we're not angry, don't worry.

Then the other thing that's not, yet possible is we don't tell you like pro BMP, we don't tell you what the severity of diseases or based on one sample. What we do do is compare a sample that you've sent in as a baseline, which is then used for monitoring and treatment, evaluation going forward, but we need to have that baseline first related position and through the eye system. And we can't tell you about very rare disease, of course.

I mean, we've, we've just launched. So, you know, if we're talking about diseases that happen 1 in 2 million, we're not going to be able to tell you yet and that that disease has occurred, but we can tell you that patient is not normal. So that's where we are.

And of course, we're open to discussions. This is a vet led company, For vets. So if you like literally get in touch, if you have ideas, if you think that there is an area you're working on that's crossing over here, and if there's something you would like us to do, then very much we would encourage you to get in touch and of course, you can get in touch and order a welcome pack.

So the next thing we're going to do, and then I'll talk about the future of the microRNAs, things to keep a lookout for as we move forward. The next things we're going to do are relatively simple. Our future work, is already underway on Yoni's disease, and we're working with SRUC research to see if we can detect Yoni's disease.

Anybody working in farm animal medicine, will be thinking that's a really good idea, I'm sure, and you'll remember from uni if you aren't working there. And we're also about to just look into mastitis diagnostics using milk samples. That's another area that I'm very keen to, to find out if or not, if, if we can or we cannot detect microRNAs in the same way in milk, I think that would open another door.

We also have this heart panel, which we're using in cats and dogs, and we think that that would be an excellent fit for the equine market. So all those settings, for example, could be very useful if we had a little bit of a heart screen and poor performance, of course, as well. So we're looking actively at the moment for collaborators to work with us on that.

Again, if you think that this is something you'd like to be involved with, and we genuinely would love to hear from you. And then we're going to do whatever we can to take this little Scottish company out into the marketplace, show, the scientific community what vets, are doing and hopefully spur on a little bit of, of human diagnostic activity as well. And whilst we're doing this, we hope to be working with NGOs and governments on, on issues such as food safety in the developing world too.

So we've got a lot ahead of us, a lot of work to do. So, where is this whole thing going though in the future, where is this technology, and I suppose, how is it developing? So I personally think that, that by developing the diagnostics, we then will gain a better understanding of the disease pathways.

I don't think we have to do it the other way around, because all we need to know to do diagnostics or to, to proceed with diagnostics is that those microRNAs are there. And consistently up regulate or down regulate at different stages of disease. I don't need to know what they do.

But once we know which microRNAs are there and working diagnostically, then we can start to think about, oh, what are they actually doing? Does this give us new drug discoveries or treatments? And I think that the, the future, I mean, it's not possible really to, to see where this market is going overall.

I think it's gonna be really a really key that we do as much research as possible. I think I'd love to know why they're in urine and blood and, and what are they doing, why are they mess, who are they messaging, assuming they're messaging, what's the function, how can we benefit more from understanding these things? And the other thing that I'm quite passionate about is how can we draw more vets into this type of research because we're a little bit behind and we really want to encourage veterinary research because there is so much potential here.

But of course, research is research and so it's much more complex than perhaps I've made out. So we do have, some, we have looked at some simple examples, but actually, disease is, the, the diagnostics is easier, the disease research is harder. I think that the mechanisms are complex and finding that pathway and being confident that you find the pathway is difficult, and then on top of that, sometimes when you've found your mature microRNA, you don't actually know which pathway it has the options of working on several pathways and genes and also several genes can be regulated by the same microRNA.

So the complexity starts to build and you can see the picture is perhaps a little muddier than I had initially painted. But there's huge potential and I think that we're on the brink, as a, a scientific community and as clinicians of a new revolution in cancer diagnostics. And I think that, that there will then be or, or perhaps hand in hand with that new treatment regime as well, where hopefully we move away from a, a generalised chemotherapy and we become more targeted as everyone is trying to do.

And I find this really interesting in terms of, what will be the side effects, for example, of the first myRNA treatment. What will happen to the inflammatory pathways that they can also involve themselves with? What will happen in terms of immune-mediated disease?

What else will we be influencing when we start anti-microRNA and microRNA therapies? And I think that now as well, in this day and age that we're currently in, we're looking at health biomarkers inside a country where we've now got this amazing testing framework. So we can now do mass population screening, which would have been perhaps a little bit of a reach 2 or 3 years ago.

So will diagnostics for humans, for example, become a little bit more remote, and what would people think of that are some of the things that run through my head anyway. And I think early detection of disease is one of the ways that we can increase longevity and quality of life in the human population as well, but also the amount of knock-on effect that we'd have for, for example, the NHS, insurance companies, the economy, and this technology really is very powerful and and I find it very interesting, to think what we may end up doing with it, in healthcare. And then I suppose we can get even more futuristic.

So genetic engineering is difficult. That's why we're not genetically engineered. It's not possible.

It's just really difficult to do. So with these microRNA epigenetic frameworks actually be the way to genetic and genetically engineer to reduce or stop ageing, to reduce disease, of course, as we talked about, but also to reduce pain, for example, I'll talk about that in a second. And will the next generation, will pregnancy, genetic engineering or screening, and will that make the next generation healthier than us, purely through the use of microRNAs?

And if so, we, if we can do this, really should we be doing this. There's all sorts of, of ethical questions. So let's look at what's happening currently, with a couple of findings that have, that are hot off the press, shall we say.

Circulating circulating circular RNAs are microRNA sponges. So this is a new finding. Circular RNAs are conserved across species just like microRNA.

And they act we think as decoys for microRNAs. They're tissue and self-specific, just like microRNAs, and they're long, non-coding pieces of RNA as opposed to short non-coding pieces and as microRNAs are. So therefore, we've now found out that circular RNAs are regulating our immune system also.

So that's a new player, a new kid on the block. And then when we talk about pain, there's been a bit of study in humans again in pain with microRNAs and multimodal pain and control is, is fantastic, but actually doesn't eradicate pain in, in a lot of cases, we still struggle with it as clinicians. MicroRNAs are altered both at the origin of pain, and then when pain becomes chronic, there are differentially expressed roles for microRNAs are differentially expressed depending on the cause of chronic pain.

So they're doing something, there, and it's thought just now that a breakthrough, an absolute breakthrough in pain management may be related to microRNA and their pathways. And then tissue engineering, and we spoke about genetic engineering of a, a full being, but actually, tissue engineering and regenerative medicine is really interesting, because remember, microRNAs control, our stem cells. So can we improve the function of tissues and can we renew cells, that previously it's not been possible to do?

And it's not yet clear how we might do this, but there's some promising results and, and how then do we deliver these microRNAs to the tissue and perhaps moving over to, veterinary medicine a little bit more again. How do we, look at microRNAs in terms of successfully growing meat in the lab. So we think that this might help with growing organs and therefore foodstuff within the lab as well because they're so essential to these, development stages.

And perhaps one that's a little close to, to the heart of some of us in veterinary medicine is debilitating mental ill health. Now, dementia, for example, neurodegenerative disorder, has been found to be linked in some ways to microRNAs, but other mental health issues have strong, strong links, to mental health. So, for example, there is a microRNA called 323AER 323A, and it's a very simple correlation.

When MER 323A is increased, depression is increased, debilitating depression. So could this be a new treatment that we've never considered before? And actually, scientists, just to give you a quick bit of background about this, scientists have analysed the brain cells of patients who died, of suicide.

And they found that within the anterior cingulate cortex of those brains, there was massive upregulation of my 323A. So they think that the target of a receptor neuroreceptor known as ERBB4, which is already known to be disrupted in schizophrenia and other mood disorders, is the target of MR 323A. And so what they then did, was they took a mouse model, increased my 32 3A, up regulated it, and the mice displayed anxiety and depressive-like behaviours, and then absolutely fascinatingly, when they donregulated it, the opposite happened.

So if we think about that briefly, with one microRNA we can control how a mouse feels. It just seems unbelievable that we haven't been working with these particles for longer. So, basically, I think that there are changes coming to the industry.

I don't want to go into this too much, it's just a food for thought. But if we can diagnose disease through urine samples, saliva samples, milk samples, perhaps this becomes patient side and it's home testing kits, perhaps there is some sort of iPhone app that goes along with this, . We've got the rise of video and veterinary, consultations as well.

So I think there's a lot that are going, it's going to change with the advance of myRA diagnostics and other, obviously, diagnostic fields, which probably some of which is needed, and some of which, which is, non-preventable, but it'll be really interesting to see what happens. And in terms of, you know, a quick summary here, we've run through a various types of, of microRNA disease and we've advanced our understanding, recently in microRNAs because we can now do assays so quickly. We've got successful diagnostics being developed, such as our new heart disease test.

And once you add in some deep tech, this is a whole new era. So our AI learning model is certainly what we needed. I think that there is definitely further research and development, required, and we have a massive lack of understanding of just what these microRNAs are doing.

And so, yes, let's, let's keep working together and see where we get to. And references if anybody would like to take a quick picture, but you can really go down a well with this as well. So, you know, there's a lot out there.

So that's us, so whistle stop tour of my area of research and I'd be very happy to take any questions. I think Jackie was maybe going to coordinate the questions. I'm just coming back here.

I'm back. I'm Eve that was absolutely fantastic. I think my, my head is expanding massively at just the potential that this has and where it can take, not just, you know, obviously the veterinary industry, but, you know, human healthcare as well.

It just, I think I need another hour just to kind of sit and digest. Everything that you just said the scope is just endless, isn't it? But I'm not going to waffle on because there's been so many questions coming through, so I want to get straight onto these, these questions from everybody, that we've already got and anybody else that's got some more, please feel free to pop them in the Q&A box.

We'll get through as many as we possibly can. So just start at the top here, . I've got, so my RNA 127 could be studied to treat lung disease and my RNA 375 to treat or manage diabetes.

Is that the case? That's kind of a question. Yeah, yeah, oh, to be honest, yes, if that was what was on my slide.

So I don't want the lung disease or diabetes, so I don't remember the exact numbers, but yes, exactly. That's the, that's the point behind this is that we can choose sometimes just one microRNA and if we can understand it or manipulate it or make an anti-microRNA or make more of that microRNA, then yes, we can. Changes the path of things like lung disease and diabetes.

That's the, that's a projector. Yeah, that's a projection. Incredible.

OK, but are the microRNAs that are currently being used and considered to be accurate for specific diseases very cell-specific in the sense that only the tissue cells where the cancer is present release them into blood, for example. Yes, which is completely different from protein diagnostic testing, which is why this is so powerful, because the microRNAs you find in the brain and the heart tissue in specific areas of the heart are completely different from those. That you find in your intestines or bones.

So you don't have this multiple, kind of, area that can come from. They're super, super specific. Yeah.

So it makes it really useful for us as clinicians. Yeah, absolutely, absolutely. Wonderful.

And I don't know if this was included in your, studies list there, but it just, there's one question saying, are there any studies of microRNAs and neurodegenerative diseases? Yes, so the, the study that I had on my reference slides was more on the depression side, but yes, exactly. So neurodegenerative diseases, which we perhaps don't encounter as often, of course, and although we don't not encounter it, we just don't encounter it as often in, in our animal work as they do in humans, it's another large area of research because we perhaps I shouldn't say we the doctors, struggle, to diagnose a lot of these diseases and clearly MRI, CT, etc.

It's quite a difficult modality to use. So yeah, absolutely, there's a lot of work and stroke diagnosis, Alzheimer's, different types of dementia, and other inflammatory problems and, and CJD and these types of things for sure. Wow, it just seems like it could be the key to everything, doesn't it?

Because just answer all the questions. Amazing. OK, what have we got here, blah blah bah bah.

How is the progress of research with infectious diseases? Yeah, so I think that more work has more funding for sure and slightly more work has been undertaken with infectious disease since the pandemic. It's, it's driven, I suppose, the idea of finding really reliable diagnostics because even with a PCR test, we know we don't pick up every single case of COVID, for example.

So infectious disease research has suddenly been funded much more than it was before, which is good, but if you think about Infectious disease, there's just so much work to do. It's like, if we all give up what we're doing now and spent our lifetimes working on microRNAs and and infectious disease, we would only get through a segment of, of what we needed to. So yeah, there's, if anybody is interested in, in working in this area, there is funding, and certainly vets are welcomed into research because we have a unique species per perspective.

So yeah, I would say if it's something you want to look into, there, there is money out there to do it. And I know you mentioned testing the milk for mastitis, so we've got a question here related to that saying, have you thought about testing for TB in the milk samples and comparing to current standard approaches? That would be cool.

That would be cool. That's a good idea. So I think that the Yoni's work that we're just on our, our, I was starting just now, we're just at the very edge of that, and that we're doing in a blood sample because that's also mycobacteria and we had hoped or do continue to hope that we may be able to do a nice easy research.

Development and trick and move that over to TB testing. And but that's a really good idea that actually perhaps the the tis and milk work we're doing along with the blood samples and Yoni might combine nicely to give us more options for TB. So, yeah, exciting times.

That's a great idea. Thank you. Yeah, brilliant.

I've got more coming through. Is anyone using whole body conditional knockout approaches in adult mouse models rather than germline knockouts, which might give a hint as to side effects of anti-microRNA therapies. Yeah, I, I bet they are, but I don't, I haven't seen any studies with this so far.

So, as far as I'm aware, antimicroAs remain a theory, which means that they are not yet published, and there's a lot of test testing going on, I'm sure, in labs, but I don't think the results have been widely published yet. Certainly not in the areas that I've been looking. So yeah, I'm sure they are though.

I'm sure this is underway because anti-microAs will be, you know, a fortune maker for someone somewhere. So I'm sure it's like an enormous amount of research will be going on in that area just now. Yeah, I'm quite rightly too.

Right, are microRNAs found in or secreted by bacteria? If they are, could a diagnostic test or treatment be developed to detect or treat bacteria that are resistant to antibiotics? Yes, exactly.

So, yes, they are, and that is, the, it is thought that microns maybe one in a number of, of developments to combat the resistance crisis that's currently facing human and animal medicine. So when we have a look at our milk s and the sti, one of the things that will be really interested to find out is firstly whether the host mRNAs are varying depending on the bacterial infection, but also what bacterial and microRNAs are doing, as well. So can we just identify bacteria based on a microRNA profile?

It all, it all helps to kind of fight that antibiotic resistance, anything we can do to avoid that. Right, the last one I've got coming through here now, does your test differentiate between overall blood microRNAs only exosomes PBMCs? I assume there might be expression differences between those compartments.

So, no, it doesn't. It just looks at what, microRNAs we capture and serum. The way we differentiate between, microRNAs that we're interested in and not interested in is because we only ask the plate to look for the ones that we see secreted and vesicles from the heart.

So that's how we cut out noise from things like inflammatory components. So yeah, so it, it doesn't differentiate in terms of the serum, only when you put it onto the plate, it captures what we would like. Wonderful, thank you.

And last one just to and hopefully a nice quick easy one coming through here now, where are available guidelines for taking, storing and delivering of samples and a price list? Are they all on the website? Yes.

So if there, there's a contact us form on our website, so we, we are slightly new age in the sense that we're working with an app rather than a paper system. So if you pop onto our website, sign up for the app or drop us an email. Then we can get in touch with you with your welcome pack, which has all of that and within it.

So it's 90 pounds for the test to, to you guys. It's a serum, separate serum sample, and it's all done through a wrap, so it's nice and straightforward. Brilliant.

Thank you so much for answering all those questions. They were coming through in droves then, but, I think we've got through most of them. So to everyone that's joined today, thank you so much for your time.

I hope you have enjoyed the session and found it as valuable and interesting as I have. Thank you again to our sponsor, My RNA Diagnostics. Thank you to Dawn, our controller behind the scenes here for making sure everything.

Smoothly today and most importantly, of course, a huge thank you to Doctor Eve Hanks for such an exciting insight into the new microRNA world and all of its potential. Thank you so so much. I hope you all have a wonderful afternoon and we will see you again soon on another webinar.