We still have a lot of people trickling in, but I know it's been a long and hopefully valuable weekend for everyone. Our next speaker is our inventor founder, Dr. Duncan Ross. He has a very unique background, which hopefully, to us, the story matters just as much as the science, right? So hopefully Duncan will delve a bit into his background in immunology and biochemistry.
So without further ado, our founder, Dr. Duncan Ross. Hello, can you hear me? Good. Thank you all for coming. It's Sunday and I know you want to get home.
That was a fantastic presentation by Jason. It actually made me aware that it's a good thing that I update my presentations from time to time so I have something new to say. to you.
So let me go through what we're actually doing here. This is the kind of thing that we see over and over again. This is on day two. He said, my friend fell off of his bicycle. And then this is, you know, a few days later, six days later, by applying these MSC exosomes to them.
This is a really strike, oops, this is a striking one. This is from Dr. Chernoff who's going to speak later. This poor girl woke her dog up by mistake, or not by mistake, but he wasn't ready for it and bit her lip off.
So he applied these exosomes and he'll tell you exactly how and that's seven days recovery and I got to meet her right before she was getting married and I couldn't even tell that this had happened to her. I mean that's really really beautiful. Dr. Chernoff's data is fantastic. He has many, many before and afters to work with, so try to stick around for him.
So I am the founder of Chimera Labs. I started it in 2012, and if you take anything home from today, just remember exosomes.com. You don't have to remember anything else. We have two projects going on right now.
We have a non-profit called Chimera Society. The idea was to be able to treat patients that can't afford treatment, and also to be able to apply for... government funding. The Chimera Labs is the for-profit and that's what we work on now. We have 27,000 square feet in Miami, Florida and I encourage you all to come visit it because I can say anything I want up here but when I take you into the laboratory and show you the cells, introduce you to the staff, it's a much different kind of an understanding of what we do.
This is our latest picture, a nice little group of people and this is what the structure more or less looks like. Every big blue circle you see there is either an MD or a PhD. What I find fascinating about this is I've kind of reached a critical mass, but I don't go in the lab anymore. I have PhDs that worked on extracellular vesicles at Yale for 12 years.
I have other people that grew placental MSCs at Celgene. Sitting around the table with them is fantastic. You know, they teach me. I'm still learning every day and even Dr. Spiel sends me a paper every single day. Dr. Spiel sends me a paper.
And you have to understand that I created this company based on wound healing. That's what I know. I'm a scientist. I know how to make things.
But the applications for them, I had no idea. And it's kind of blown my mind the way people are using them today. So this is my father. He's a scientist, or he was a scientist, at NOAA and University of Miami. He used to fly into hurricanes and study wave heights.
Well, he got myelodysplastic syndrome in the year 2000. and I had to come back to being serious and applied to graduate school and got into the biochemistry department. But I had six years with him from his MDS progressing to leukemia. Six years to read blood and the biology of blood and marrow transplantation and try to understand how to treat leukemia, which as anyone in here knows if you try to treat cancer out of the box you have no idea what you're doing.
And when you read the papers, which you know a third of them are actually incorrect, you don't know how to parse what you want to do. So in fact we we kept going to chemotherapeutic methods that the local hospital was telling us to do, when we should have gone to bone marrow transplantation. And by the time we understood that we needed to do that, they wouldn't treat him anymore. I had him at Harvard. I had him in Seattle.
We missed our chance. So I am now published in those papers, in those journals, Blood and the Biology of Blood and Marrow Transplantation, because I love transplantation, and we know a lot about it. I mean, the first transplant was in 19...
The function of regulatory T-cells is really carried out by a protein called TGF-beta-3. TGF-beta-3 will turn an inflammatory CD4 T-cell into a non-inflammatory CD4 T-cell. So think about it like this.
If you have arthritis and a CD4 T-cell is blowing up a knee and you put TGF-beta-3 in a dish with that, it will stop that inflammation, just that one protein. So knowing that, and knowing that what was going on in transplantation at the time, I knew that people were using mesenchymal stem cells to suppress graft-versus-host disease. In my laboratory, we were using regulatory T-cells. But companies were showing that with 14 million MSCs from an allogeneic donor, you could stop chronic graft-versus-host disease.
Now that's a very robust inflammation. So the inflammation that we're worrying about here... arthritis is much less, it's much easier to deal with.
So at least you know that with 14 million MSCs you can do something. So the MSC starts to become sort of a golden goose. An MSC is what you should think of from now on as the center of your body.
You used to think of your heart as the center, no the center is your MSC. As you're growing, that's why it's from the mesenchyme right, as you're growing it's controlling the growth of the organism. Something is telling a kidney cell to grow, something's telling a liver cell to grow, but the liver cell doesn't care about the cells around it.
So this cell sits in the vasculature and secretes growth factors and it also secretes anti-inflammatory factors. It does that only when you're young. So if you look at this, it's hanging out on the vasculature, it's secreting exosomes right down into that vein or artery. And now those growth factors are proliferating throughout the entire body. So knowing this, and remember in 2012 people were talking a lot about stromal vascular fraction, I decided that I was going to apply for an IRB to try to treat COPD using 60 cc's of fat from donors.
I treated many, many patients with this, probably about 250 if not more. Remember, I'm a scientist, I work with many physicians, so I've got data in from all kinds of different directions. The one issue I thought is you have to charge these people, somewhere between 6,000 and 13,000 people, to take their fat, have the surgical time, isolate the stromal vascular fraction, and treat them.
Now, almost every time, I would say 60% to 80% of the time, the patients would improve. Their FEV1 numbers would go up. The issue was that after six to eight months, they would lose the effect. And now sometimes it's because they were cheating, because they would smoke, or cause some other kind of inflammatory insult. But it just wasn't making sense to me that so many of the patients relapsed.
So I thought, okay, well I know that there's 150,000 mesenchymal stem cells and 60 cc's of fat. Just for your knowledge, there's also 50,000 mesenchymal stem cells and 60 cc's of bone marrow. That's still not a lot of cells.
So knowing that Osiris and Mesoblast were using 14 million cells, I thought, okay, I'm going to expand these patient cells to 7 million cells. I'm going to treat them with their own cells. That should be better.
I thought that was going to be my business. Well, it didn't work out quite that way. Because I would treat them with their own cells, and many people, absolutely it was better, but some people's cells I couldn't even get to grow. If you look in this room, your age means nothing. The efficacy of your cells is entirely different.
I had 65-year-old men... whose cells grew like an 18-year-old girl, and I had 70-year-old patients whose cells would not grow, no matter what I did. But I did know that there were experiments that showed that if you connect the circulatory system of an old mouse to a young mouse, the young mouse will get older, and the old mouse will get younger. Now, the old mouse getting younger makes sense, because I just showed you that the MSCs secrete exosomes out into the vasculature.
But the... young mouse getting older is pretty interesting. Why would that happen? The reason that happened is because you have senescent vesicles being secreted by the old mouse that are now going into the young mouse and killing the young mouse.
So there's actually a very interesting physician that I know named Dr. Haas and he is leukaphericing the patient or aphoresing the patient, returning all the cells back to the body but taking out fifty percent of the serum such that you remove these senescent bodies. I think that's a fantastic idea. I want to do it to myself and my mother.
So think about it like that. So one day I had this 70 year old patient who was actually the wife of the plastic surgeon that I was working in. I had built my lab. in his kitchen inside of his clinic. So it's my tiny little hundred square foot lab.
So I'm telling him that this expansion of cells is going to be a big business and I can do this and I had this 18 year old paraplegics patient on the right whose cells were growing quite well like wildfire and then I had his 70 year old wife on the left and it wasn't growing at all. I didn't know what to do because if I couldn't grow his wife's cells I'm in trouble. So I did everything I could. I changed the medium, I changed the oxygen concentration, and they wouldn't start growing.
So finally, I'm about to throw out the cells on the left, because I don't care anymore. But I thought, well, I know about that experiment with the mice. Let me take the supernatant of the 18-year-old, which has proteins and growth factors in it, the paracrine effect, and put it on the 70-year-old cells and see what happens. Because it's not ethical, but I'm going to throw the cells out anyway. So when I did that, the 70-year-old cells started growing.
That's why right away I was like, oh my God, I've been throwing out this medium. And a xenofree chemically defined medium is 10 times more expensive than a medium that uses fetal calf serum. And I didn't have a lot of money at this time, so I was like, I'm throwing out the gold.
The gold is in the growth factors. I don't need the cells to cause growth. And so that's where this exosome understanding started.
I did not invent exosomes. When I started looking around for pericardial effect, growth medium, condition medium, China and Singapore was really a leader at that time. People understood exosomes. So we now know, or I now realize, that I don't have to use the cells because the problem with the cells is that they persist for a short period of time. If you take a cell out of its niche and you put it into the vasculature, or into the circulation, it's going to die.
Now while it's dying, it's going to secrete exosomes, it's going to have some effect, but it is still going to die. That's the issue with umbilical cord use that's been going on for the last couple of years. Everybody knows that there is some positive outcome, but there's a lot of negative outcome because the cells die.
Now if I'm going to use it as a therapy and have to expand your own autologous cells, it takes me six weeks to do that, so it's not going to help if it's an acute injury. Third-party cells get rejected. That is the issue with cord blood.
That's why I never wanted to use that. And the growth of these cells is really not approved for people. The other issue is that old people's cells are old.
Dr. Sanders referenced lactose tolerance. We're not supposed to be lactose tolerant past three years old. But we all have the gene. We pass it on at 20 years old and we continue. So the lactase gene has been epigenetically silenced.
That's why we become lactose intolerant. But every growth factor we have has the same result. And that happens by design. So we don't become Andre the Giant.
We need to stop growing at some point. But regenerative medicine, what it's really going to be is opening those growth factors in a controlled fashion such that we can not only stop aging, but rewind aging. Right now, I feel like we're stopping aging, but my goal is to rewind aging.
And I know that there are five other companies Using different methods of applying the Yamanaka factors to create induced pluripotent stem cells to try to rewind aging. Rewind aging by two days and let aging go forward one day. Back two days, forward one day.
In that fashion you don't get a tumor and we already know how to rewind aging. I think that is pretty exciting. If we can all stick around for ten more years, I don't know what's gonna happen. We're really gonna expand.
So it had been shown at this time the condition medium could have the same effects as the use of cells in an emphysema model, for instance. And I was working on COPD at that time. So that's encouraging.
So that was in March 2014 that I realized that the supernatant, the conditioned medium, was all you really needed for regenerative medicine use. Then in October of 2014, I went to a party, and my friend was drunk. It was his party.
And he threw water on a gasoline fire, and he burned his face in that fashion, second degree. So the next day he's very upset and he's black so he has a possibility of keloid scarring, hyperpigmentation, all types of things. He's very very upset. So knowing that I had been creating this condition medium and I was now freezing it instead of throwing it out, I convinced him to allow me to apply it to his face three times a day for seven days.
So if you look at look at the sides of his mouth right there you see how it's kind of turned down he looks not very happy. We'll look at him seven days later. So he's kind of smiling and then look at him six months later. In effect he got a free peel, facial peel.
Look at the size of the pores in the new skin versus the old. So what we'd actually done is by taking young growth factors and young messenger RNA, we allowed him to make young new skin and that was really my first visual understanding of what was going on here. What the power of not using cells and using exosomes was. So what is in that condition medium?
I was in graduate school for a long time. I actually did the whole PhD program of biochemistry and then had to switch to immunology and do that whole program too. I never understood how a protein, which is so labile, that when I would express a protein I'd have to run to the bench to try to get the assay done before the thing was not useful because it would denature so quickly.
There was no way for me to understand how proteins in the supernatant of conditioned medium could be useful in any way. And in fact, they're not. What's happening is that the body secretes these 100 nanometer lipid vesicles that protect the proteins but more importantly protect the messenger RNA of that protein. So with that one messenger RNA you can create millions if not billions of proteins.
So if the half-life of VEGF is four hours the half-life of the VEGF messenger RNA is four days. So in fact what the body does is send the message to the area. and then express the protein in situ. So having grown cells and used so many cells, I started understanding what the amount of volume of the supernatant was that I had to use to create an effect.
It didn't matter how many billions of exosomes were in there, I didn't know. I did know a protein concentration of what was in there. So when you think about these exosomes, it's very confusing because somebody says, There's a billion over here, there's 300 billion over here.
Well, I'm going to get to that. But if you really wanted to be a stickler, if you have a 70 nanometer bone marrow exosome and you have 120 nanometer, 130 nanometer placental exosome and you have twice as many of the bone marrow, you don't really have more because the volume of a sphere is four thirds pi r cubed. So the larger the exosome, the much...
increased amount of proteins and messenger RNA you have in there. So don't get bogged down by numbers. Get bogged down by dosage and efficacy. If a physician tells you how much to use, that means something. So where do you find exosomes?
You find exosomes in blood, but they're usually older exosomes. You find blood in urine, and I'm very interested in that because we're creating a diagnostic portion of our company because when you To create so much characterization equipment and protocols, it would be almost a waste to only use it on our exosome. We can develop new methods of diagnostics for prostate cancer, for kidney disease.
We can pull all of these exosomes out of the urine, assay them, and then understand what's going on in the body. And that's really where the whole field is going. There's a lot of investment in that right now, and it's very exciting. There are exosomes in amniotic fluid. They're a different type of exosome.
They're an epithelial type of exosome. And we actually make amniotic fluid at Camaro Labs. I love it.
But amniotic fluid contains a different exosome than a placental MSC. And it's also impossible, as far as I'm concerned, to isolate exosomes in a purified manner from amniotic fluid. And I'll get to that in a minute. You can get exosomes from bone marrow.
just like you can get exosomes from blood. I mean, when you take a bone marrow aspirate at 60 cc's, you know that about 50 cc's are blood, right? Some of it is bone marrow.
But if you spun that down and took the serum, there would be a number of exosomes in there. And those would be useful, but the older the patient, as Dr. Sanders spoke about, the older the exosome. You can find exosomes in milk, but you can't sell milk as an exosome.
It's milk. An interesting thought is that the eight zip codes that most of the Army recruits come from that actually pass the test are in Wisconsin. They're mostly children of dairy farmers because they drink unpasteurized milk. So when you drink milk you get cow exosomes into your circulation and you express cow proteins for a period of up to a month and that's why they're so big.
So you can think of that next time you drink milk. What we do at Khmer Labs is we create a cell culture condition medium that you can get exosomes from and you can also get exosomes from tumors and this is the problem with using the word exosome. You really don't want to tell your patients you're using exosomes because the first thing they do and I've had this issue for is they Google exosomes.
And a major part of research now is that metastasis is really a cancer cell secreting an exosome with the detrimental DNA in it and that exosome goes to say the lung and then it transforms the cell in the lung and people get very scared when you talk about that. It's fascinating that that's how metastasis is working but the word exosome is probably not good for treating patients. If you wanted to say placental growth factor you know that would probably work.
So the benefits here are you may know that the FDA has come down very hard on cells in the last year. The reason for that is that there is no way to control the contamination of cells especially cord blood that's being taken in a in an operating room. An operating room is far from sterile. The sterile field is not sterile at all.
I could never grow cells on that. So when we have exosomes now Even if the culture was contaminated, I could filter out the bacteria using a 0.22 micron filter. I can then test the product for sterile netto toxin before I let it out.
I can concentrate it to make it more strong, if I wanted, more potent. It's not going to be rejected by a third party because it's too small. It can't be seen by the host. It also doesn't, if you're taking it from a zincable stem cell, it's not going to have MHC class 2 on its surface. Some of them do have a little bit of MHC class 1, but not enough to really create an immune reaction.
And it can be frozen, not cryopreserved. Cells are cryopreserved in DMSO because they're cells. But these exosomes are just proteins and messenger RNA.
So that makes it a lot easier to handle them. I keep them at minus 80. You can keep them at minus 20, which is the regular freezer above your refrigerator. So there's no stress there.
I can also make different types of products. We have an ExoGlo Pro that has more product in it. We have an original ExoGlo. You do not want a stronger product. Do you want a Viagra that's ten times stronger than a Viagra?
Nobody wants that. We have a product that has a dosage and treating patients is something that is your art and depends on the weight of the patient and you don't want to overdo it. One of the funny things about treating patients, even from before when I was doing stromal vascular fraction or cell expansion, is men would come in and say well I didn't get any effect I don't have any more energy.
I said okay nothing happened? No, nothing, nothing. Well alright maybe one thing. What?
Well I'm waking up erect now. Waking up erect is actually the way we dial in the dosage. Everyone reports that. So You can quantify these types of things.
In my opinion, I only take our exosomes once every three months because it bothers me to take it more often than that. I don't need it more often than that. Dr. Spiel is going to show you some data that's going to be really fantastic.
We've started testing the DNA methylation age of a number of markers from before we treat until after we treat and he has data from that now. So that's fantastic. So what's happening is the MSC, think of it as say the quarterback, it's throwing out The ball which has the message in it which is arriving at the receiver and then it's allowing that receiver to do what it needs to do in the environment that it's in.
So one messenger RNA can make billions of proteins, one micro RNA can control the expression of a protein inside of that cell and so that it's the manner of cell communication. An interesting way to think about is what came first the chicken or the egg what came first the virus or the exosome. Viruses and exosomes are the exact same size. What I'm going to talk to you about in a minute is that I create our exosome according to FDA guidances for vaccine production, because that exists.
So we're creating proteins in situ, so something easy like VEGF is fine. But why are the exosomes working in a regenerative medicine capacity? If any of you have done a lot of regenerative medicine, you'll find, and even the Mayo Clinic had this at Toby, some patients don't get better for eight months after they are treated. How is that possible? The reason that that happens is, look, whatever therapy you did to them is gone.
People get a first pass effect, they get a second pass effect, and then they get a long-term third pass effect. That long-term third pass effect is because of epigenetic remodeling via proteins that are called histone acetyltransferases. or histone deacetyltransferases. They're proteins that can put a methyl group on a histone which will open the histone and then the DNA can be expressed. I know through messenger RNA sequencing that we have those proteins in these exosomes.
So you are remodeling the DNA and allowing the cells to start working as a younger individual. When I started this, even before I was... the exosome out of the medium.
I gave myself 50 times the dose that you're supposed to that we now give. Now I'm 43 now, 37 at that time. I didn't see anything, whatever, I didn't have any more energy.
But eight months later, my wrinkles went away. Now, fortunately, they came back. Because what I'm saying is we have been worried about tumorigenic effects since the very beginning. And it turns out that exosomes are now demonstrated to be anti-tumorigenic.
But the remodeling of the DNA expression and the regeneration of tissue takes a while. It took you nine months to be made. I don't know why you want your knee fixed overnight.
But it is striking how many patients see these results. results months and months later. The way we're starting to think about it now is treatment once a year or something to that effect.
So like I said, we're not growing anymore, right? The reason we're not growing is because the controller of cell, of body growth is the MSC and it has turned off the requisite growth factors. So we have about, let's pretend, 110 approximately proteins coming out. A baby's mesenchymal stem cell secretes about three 300 factors, quite a bit more, because it needs to grow. So it does a couple of things.
Baby cells, they grow, they're anti-inflammatory, and they're non-tumorogenic. On the count of three, please, I want you to say young. One, two, three, young, okay? Because exosomes are not the conversation right now.
Young exosomes are the conversation. And it is imperative that you understand that, because you can get an exosome from anything. You can get exosome from my old cells.
It's not going to help you. And the reason that if you've ever done stromal vascular fracture or regenerative medicine, the patient comes back and says, you did something wrong. It didn't work. It was not your fault.
It was 100% their fault and their cells. But how are you going to explain that to them? And I went through that with thousands of patients who were working with my partners, and I just got tired of it.
I was called a fraud twice, and it really pissed me off. I mean, I've been published in a lot of journals. I've been in school almost my entire life.
It's not fair. call me that. So Doug Spiel coined this phrase, exosomes are the penicillin of our generation, and I think they really are changing things.
So let me tell you how I do it. And again, this is the way it's done. I didn't make this up. This is the way I'm required to do it.
It's how Humira is made. It's how Botox is made. You always have a cell bank that is characterized.
I take a placenta, I dissect it, and then I throw it out. out that placenta is only in my hands for 24 hours. I don't have a placenta that I've been hanging on to for four years.
I isolate those cells. I grow them for a period of eight weeks. While I'm growing them, I'm able to characterize them.
I'm able to test for any virus that is available that I can test for, say Zika, Epstein-Barr virus, things that I don't have to test for. I also am doing something important is that I'm diluting whatever was in the placenta. So when a placenta is gross, okay, placenta is full of blood. It's like a sponge full of blood.
Blood gets everywhere when you're dissecting it. You don't know when you get that placenta if it has AIDS or not because you don't have the testing back yet. So it's just not a fun experience. So we isolate the cells. We grow them for eight weeks.
They now haven't seen the parent for eight weeks. Any virus that was in that original placenta will have been diluted out. Any anti-AB antibody that was in that original placenta will have been diluted out.
And then we get to the end of the day. get that condition medium that I treated that burn patients face with. At this point I can sterile filter to make sure that there's no bacteria in there.
Now this is the way the literature talks about it's not exactly the way I do it but the point is the same. Your centrifuge in your office does about 1200 G. With an ultracentrifuge you can actually pellet down exosomes at a hundred thousand G for about eight hours. When you get that exosome down to the the bottom you can remove the supernatant, which would be the conditioned medium, and you're left now with a pure exosome product.
I resuspend it in 0.9% saline. The International Society of Extracellular Vesticles definition of an exosome is an isolated exosome. You cannot call whole amniotic fluid an exosome.
You can't call milk an exosome. You have to isolate the exosome. And why is that so important?
Because the conditioned medium is definitely a drug. There's dexamethasone. There's recombinant growth factors in there.
There's nothing natural in there. So you have to remove that condition medium. We then quarantine it, test it, and try to figure out how many exosomes we have.
And that's a lot harder than you might have... imagine. And in fact, I've been working on it since 2014. So as I said, if somebody's going to argue about how to make a drug, Humira, Botox, all of these things are made from Cultured cells that's how the FDA wants you to do it and there's this guidance here If you're interested in you can go read it if you want to make a product from a same-day primary cell which is what taking a placenta and isolating exosomes out of it immediately would be You have to use a specific pathogen free population of animals that have lived in a sterile environment for multiple generations You cannot just take things out of the environment and put them now. There's no woman that has been in that situation They require that you culture the cells and you have a characterized cell type that you're making your product from.
So exosomes is a terrible word. If you look at these two teams here, both of those helmets are exosomes. One is a better passing game, one is a better running game, but they're different products. Try not to think that one exosome or more than one exosome is the thing. So in the world of academia, this is still the way people do it.
The protein concentration is how you tell how much of an exosome you have. That's it. And exosomes are so complicated that there is a method called A280.
You hit them with A280 wavelength light, and you should be able to see the protein concentration. That doesn't work on exosomes. Now, when you're trying to figure out how to work with exosomes, and the typical methods of working with proteins don't work, it can be very, very confusing.
So we use a Bradford assay to try to come up with a repeatable concentration of exosomes. And we still do it that way. That's how we characterize it.
every vial. I go on about numbers of exosomes, billions of exosomes. It doesn't matter. The protein concentration matters.
The number of exosomes was for me, so I could create a repeatable product, so I could understand things. But this is really the way to do it. Like I said, you don't want a stronger Viagra.
We have X number of concentrations. We know exactly what those do. You don't want to use the pro on a little old lady that might get sick from it.
You want to use the weaker product, or the stronger. the regular product. So the second way to characterize exosomes is by flow cytometry and we do that as well.
What's difficult about that is you attach an antibody to the exosome and when you look at the data it's confusing. As a scientist I don't understand this data very well. I see that nobody in the world knows exactly what's on the surface of an exosome. Every little lipid vesicle that's 100 nanometers is an exosome.
It might have a have CD63, it might have CD9, it might have CD81. If you go to the real conferences, nobody has definitely determined what is the therapeutic type of exosome yet. So we're all kind of shooting in the dark.
So if I change my mind or I say something's different, well, we hadn't seen an atom when we knew that atoms existed. We knew about gravitational lensing before it was proven. This is really a novel science, and it's really hard to understand.
So flow cytometry, I was actually yelling at my staff. this data isn't good enough, I don't understand it. So we continue. You can do protein expression. And this is an assay I did in 2017. And people get mad at me for why I still show it, because I don't want to do this assay again.
It's a waste of my money. It is interesting in one respect. You can see the difference in expression of growth factors in amniotic fluid versus placental MSC exosomes. And the bright side is that they're different.
So you have two two different tools that you can use for knees, for inflammation. Amniotic fluid is temporarily anti-inflammatory, and it will help the production of collagen. Placental MSC exosomes are permanently anti-inflammatory.
Now, if you look carefully at these numbers, some of them are higher in MSCs, right? The fact that there's 300 billion particles in the amniotic fluid does not mean there's 300 times the growth factors. The number of particles, and I used to say exosomes, is misleading.
I don't know. and you might not want to bog yourself down in it. The reason I don't do those growth factor assays anymore is because now we do mass spectrometry. Mass spectrometry I don't have to worry about. You don't put antibodies in there.
The antibodies can't get through the surface of the exosome, so growth factor panels are misleading. They're not valid. Mass spectrometry absolutely is. The reason that I have to do this kind of data is because we are applying to the FDA in January. Because of the recent uproar, we moved it up.
I would have waited probably another six months. But we have... all the data. Love the FDA to come tomorrow and say, okay, now I don't have to ask you for a meeting, let's just go.
So, the other way to really characterize repeatability is, I don't just have one placenta. Let's say I have to get another placenta. I have to characterize for the FDA that that new placenta is equivalent to the old placenta and the dosages that we came up with. Messenger RNA sequencing is a fantastic way to do that.
What I love more about it is that I get everything that is in the product. So almost every single day somebody asks me if I can help this or I can help that or this person has this DNA mutation and I get to go to my PhD and say do we have this protein? He goes right to the sequencing, looks for the sequence, says yes we do or no we don't. We do it every day.
It's an invaluable resource and this is really the way to characterize what your exosome is. That's also how I know we have telomerases in there and the histone and acetyltransferases and such. In 2014 when I was trying to figure out what I was doing, people did understand exosomes and the one way to see them was with electron microscopy.
Electron microscopy is hard. You have to decide what kind of metal you're going to use. It can take you weeks to do it. It's not something that you could use on a batch to batch basis, which we make three batches a week, so I have to have something that can move quickly. Electron microscopy is not the way to characterize your exosome.
But I have taken the time to characterize the product and this is electron microscopy. of the Chimera exosome. This is all new data from one of my scientists. This is atomic force microscopy of the exosome.
This machine can actually see molecules, individual protein molecules and phenol rings in those molecules. So we're starting to really believe what we're saying here. This is more atomic force microscopy. This is the The outer edge of the exosome up there on the right and the whole exosome on the bottom. This is a beautiful image.
This is focused ion beam microscopy. Look at that exosome. It looks like a little crystal.
You're actually seeing what the surface looks like and how those proteins are kind of amorphous on that surface. So one of the issues with characterization has been that this nanosite has kind of become... A standard of everyone saying how many exosomes they have, and I have contributed to the misnomer of that.
What you see on a nanosite is this, and while in my case those are exosomes, if you put sugar in water and put it on this machine you would see the exact same thing. If you put medium on there you'd see the same thing. If you took a placenta and ground it up and put it on there you'd see all these little dots, and you would say, oh those are all exosomes.
They are in no way exosomes. They're particles. And even though I purchased the laser to try to look at the surface of these particles, they're moving so quickly that you can't get any data on them. We can see that we have a particle of basically one size, which is encouraging, that's what you want to see, but the data really isn't that great. I mean, you can see, I can't see the difference between the background and the data here.
If we knew that we're trying to characterize exosomes by these surface markers which you can see by Western blot, I was trying to see it by nanosite and I couldn't see it. So what it's doing is bouncing a laser off of the particle but again the particle can be anything, it can be a grain of sand. But I wasn't using the nanosite to sell you, I was using a nanosite to make a reproducible product for myself.
If you look at the exosomes with a regular microscope you're gonna see an empty hole there, you're not gonna see anything. Well, I've lucked out in that the hardware and the science has progressed to a point where I can better characterize what I'm doing right before I'm going to the FDA, so I don't walk in there like a jerk and not know what I'm really doing. What you're looking at right here is a mitochondria. Those are antibodies attached to the surface of mitochondria. Now, I know you've only seen it in electron microscopy before, sort of a flat slice.
But look at the resolution of this microscope. When they chased me down at this conference and showed me this, the hair stood up on my arms. I've never seen that before.
It's amazing. So then they showed me that I could track those little particles running around there. Those are exosomes being taken up by cells. And now the next picture that I'm going to show you is the first light-based image.
Remember the electron microscopy is electron, it's not light. This is actual photons. It's the first light...
beige image of an exosome in the world. And that's the Chimera exosome right there. So now, when you looked at that flow cytometry and you didn't understand which was CD63, CD81, that's because that exosome right there has...
CD63 and 81 on it, but the one to the left doesn't, or it has mostly CD81, a little bit of CD63. Exosomes are a heterogeneous population. They come from different parts of the cell, and they're all going to have different surface markers.
That's why trying to characterize them by surface markers is never going to work, and everybody in the industry knows that. It's a problem, but it's not really a problem if the product works. It doesn't really matter what's in there if it works. And if we're able to sequence my messenger RNA and know what's inside of there, we can do it. Right now I'm looking at surface markers.
there, but with this machine, I could put an antibody for VEGF in there, and I could see if one individual protein molecule of VEGF was inside of that exosome. That would be a method of characterization of an exosome. So because of that, I'm trying to get away...
from talking about billions of exosomes. We're creating a ROS unit, which is going to be the number of exosomes, the size of the exosomes, the protein concentration, specific growth factors that are in there, and turn that into a formula and come up with an efficacy unit. That's how we're going to proceed. So just for your knowledge, when you inject exosomes IV, they end up in the spleen and liver, which is a benefit over cells, which only ended up in the lungs.
But for COPD, that was fantastic for the lungs. And just so you know, the reason that lungs get stuck in the lung, or cells get stuck in the lung, is because the lung is a site of extramedullary hematopoiesis. When the cells get there, they think they're home, and they extravasate into the lung, and they cause anti-inflammatory effect, and that's functional in COPD. If you look on the bottom, on the second line there, if you have a kidney injury, a kidney injury model, the exosomes will end up in the kidney.
That speaks to the fact that the exosomes go to the spleen, get taken up by macrophages, then the the macrophages traffic to the kidney. So exosomes can home in a way, even though they're not smart. They home using endogenous cells that are in the body. So 2020 is an exciting year.
I'm super excited. Like I said, I've been spending since 2014 trying to get this data together for an FDA application. And the deadline is coming.
November 2020, we're all supposed to have applied to the FDA. And otherwise, you're not supposed to sell your product anymore. So. A few days ago, a letter came out. I've been successfully selling exosomes since 2014. Never had an adverse event.
Now there's a lot of exosome companies. I specifically know one that I think does things the wrong way. A few weeks ago, December 6th, this letter came out from the FDA discussing five adverse events.
Those were not my adverse events, but this has now happened to the exosome industry. So they put out this letter. You do have a duty to warn the public about an exosome that's causing adverse events. This letter does not say you can't use exosomes. It says that there's a couple out there that are bad.
It also says they're trying to work with people and that there are none that are approved. I'm not sure why they said there are no FDA-approved exosome products because there is an FDA-approved exosome product. That's sort of the issue with the FDA.
I love the FDA. Am I interrupting? with them has been fantastic but sometimes they write things so this is the November 2017 guidance not a draft guidance it's a real guidance and I was inspected on October 24th 2017 so this came out two weeks after I got inspected and when they were inspecting me they were just harassing me about whether I was using cells or not I don't use cells the whole point of my company is that you don't need cells you need the growth factors so they published this in November 2017 says secreted body fluids such as amniotic fluid are generally not considered an HCTP.
They're not 351 and they're not 361. Now, you know there's a ton of exosomes in amniotic fluid. So basically, that sentence is saying that exosomes are not a 351 or a 361. It says cells from secreted body fluids are considered an HCTP. So you would have to remove the cells from the amniotic fluid to have something that wasn't a 351. Then it also says the following articles are not considered HCTPs.
Secreted or extracted human products... such as milk, which has exosomes in it, collagen, and cell factors, which is what exosomes are. So that is what she was thinking when she came to me.
If you're worried about what's going on or lab or anything like that, you can look up everything. You can look up my inspection. You can look up my tissue establishment registration.
And if you look at that registration, you see how it says process there? That's the one that gets me in trouble. That's the one that brings the FDA to me, so I'm inviting them to come. So if somebody has a tissue establishment registration that's just store, label, and distribute, they're a medical marketer. They're not a laboratory.
So this is the results of the inspection report. They inspected ExoGlow on 10-24-2017. That's when they expect the ExoGlow product.
And there were no 483... ...inspectional observations. So that's important because they took my process.
They made me tell them exactly how I was making the exosomes. They took it. They kept this inspection report open for two years while they went around to every clinic I deal with. and everything else to see what claims I have made, which I've never made any.
And eventually they closed it. I received the disclosure on 8-22-2019 after being expected in 2017. So this is by no means an approval, but this is what I know. So everybody wants to be an FDA pundit and they never work for the FDA.
The FDA says this, the FDA says that. The FDA is going to come get you if you are doing something wrong. And they're taking a risk-based approach to regulation until November 2020. If I don't have my product in by November 2020, that's on me.
But in the meantime, we didn't even know what an exosome is. I'm telling you, we're still trying to figure out what it is. And I have the best hardware in the world to characterize it.
My PhD, who worked on extracellular vesicles at Yale for 12 years, says we have more data than anyone in the world. So how is the FDA going to deal with these type of things? If you hurt somebody, you're going to be in trouble. If you're interested in the inspection classifications or seeing what I just showed you, you can go to the inspection classification database and look for it. If a lab hasn't been inspected, it hasn't been around very long because when we register, we understand that within two years they're going to come inspect us.
The reason I take this so seriously is because I don't want to go to jail. I'm the only person responsible for this laboratory and this guy went to jail for ten years. I take it very, very seriously.
My quality control has done phase one and phase three applications for monoclonal antibodies. I have four people just on that side. I have 40 people in total working at the laboratory and we just take it very seriously. I don't want anything to go wrong.
There is no reason why you should sign a contract. I mean you are practitioners. I'm telling you a new exosome is going to come out. An exosome that rewinds aging is going to come out whether I make it or not. Why would you want to sign a contract with somebody?
That's just a marketing ploy. so that somebody can sell their company. All of these people are rushing to make as much money as they can by November 2020 so they can retire. If you know how much our lab makes a month, but also spends a month, if I wasn't doing it the right way and spending all that money, I could retire in 10 months, 12 months.
But that's not what we're going for. I want to make that exosome that rewinds aging. Lastly, I want to talk about cancer because this is really exciting. And actually two nights ago somebody told me something I hadn't even thought about. I'm trying to get FDA approval for a certain indication, but I could probably get it a lot faster on cancer.
Because if you think about pancreatic cancer, it's usually a p53 mutation or p10 mutation. It's a tumor suppressor mutation. So what that means is, pretend you have a tire and the tire is flat now because it has a p53 mutation and it is expanding uncontrollably.
Just because the DNA is mutated, doesn't mean you couldn't put the real protein back in there and the protein would then control the growth of that cell. So we can surpass or suppress this DNA mutation by putting in an exosome that has the messenger RNA for p53 such that p53 will be expressed and now the cell will act normally. At the last ISEV meeting in Japan a few months ago, there were five different labs working on this.
We can stop if we know the genetic profile and we know that we have that protein in the exosome, we can stop the progression of the tumor. And if you give a tyrosine kinase inhibitor, you will even shrink the tumor. So this is a new type of a therapy.
And I've actually already seen it work. I've seen the laboratories that make it work. You take a myeloid leukemia, you dump an umbilical cord exosome in there, the myeloid leukemia stops growing. And that's why. So that's really fascinating.
So not only are we not worried about... tumorigenesis now, we think they're anti-tumorigenic and that should make you very excited for the thoughts. One last thing about that is it turns out that in a solid tumor in the vasculature there are 200 nanometer holes because the tumor needs so much blood and these exosomes are 120 nanometers so you can understand how they can preferentially get into the tumor. That's the end of my talk. If you want to find me again exosomes.com is all you have to think about.