Tissue Rejuvenation via Plasma Dilution | Irina Conboy, UC Berkeley

5:02PM Feb 24, 2021

Speakers:

Lynne

Jose

Robert

Raphael

Joshua

Jean

Christine

Alexandria

Karl

Keywords:

procedure

proteins

aging

cells

blood

brain

senescence

question

people

work

clinical trial

young

tissues

animals

exchange

younger

improved

factors

published

rejuvenate

Thank you, and welcome everyone to four sides health extension, and a group that is sponsored by 100 plus capital. And today, I'm super, super excited to have even a combo here, who's gonna speak about tissue rejuvenation via plasma dilution. And a winner was a while back at co chairing, and health extension, technical competition that we had. And so I'm really, really excited that you were able to join us again, I know that many, many people here are very excited for what you have to tell us. I'm going to kind of like backtrack all the questions for later and just have you start so that people don't get too overexcited. And then I'm going to take questions here in the chat as they're moving in. And I'm going to post your bio biography and everything about this group in the chat so that I don't take any more time away from the presentation. Thank you, again, from all of us here. And I think you can go,

yeah, thank you guys for inviting me. So let me share my screen. Yeah, I hope you can see it. And so my talk will be about plasticity or wage, and how we can prevent attenuate and possibly even reverse age imposed diseases as a class, not one by one, which was not successful, but altogether. So primitive bill is speaking an antibiotic against aging, which you can see here is exactly what I see from my office at Berkeley is pretty nice courtyard. And unfortunately, I was not able to go there because of the pandemic for quite some time. So this slide is introduction into how we understand or rather do not understand the causes of aging. And you know that with chronological age increase, there is a dramatic increase in our organ deterioration and disease, which at some point becomes catastrophic. But interestingly, this chronological age is very different for mammals that are similar to each other in many parameters that we consider to affect aging. For example, rats live only three years and squirrels live 20 or 30. They are very long lived mammals. And even though that squirrels have similar size, and metabolic activity, in fact, if anything, squirrels have higher metabolic activity, higher metabolism, dendrites, they are mammals, and they have very similar diet. But yet, squirrels live 10 times longer. And so by the time the squirrel is three years old, it's a very healthy animal, but there are at three years old will be very sick, and really, really aged animal. And so TAs, first of all, it tells us that we do not really understand what controls the rate of mammalian species aging, we don't even know they have some theories. And secondly, that it is not simply the entropy or the rate of damage that controls because rats and squirrels really live in the same entropic environment. So, perhaps it is the capacity for repair. So, damage is something that we cannot change, but repair or efficiency or repair we can change. So here is another cartoon viewpoints on what a mansion See, if you look at the young person and an old person, the degree of damage will be the same. In fact, an old guy who takes care of himself or herself perhaps has less damage, if the half how to diet and wear sunscreen as compared to young people who may be basking in the sun and go clubbing every night and so forth. However, the rate of repair is significantly higher at at the young age. And so, perhaps, if we can improve repair an older individual, we can then rejuvenate them. So, this is kind of the general conceptual introduction to my talk, explaining what are we doing and why we believe that we cannot really chase down the entropy itself and reverse it that every single minute detail and to be perceived, but we can attenuated and offset that by improving render generation and that is the key to longevity. So I will speak a little bit about our experimental systems. You look at mesodermal rejuvenation and the dermal rejuvenation and ectodermal rejuvenate And those are the three germ layers from which our bodies are built. So we really start with messenger, mcdorman and the door, and then those differentiate into various organs and tissues. And so it is important if you look at multiple tissues in the nation as a class to make sure that all three germ layers become rejuvenate, for muscle for message on the look at muscle, and we'll look at fibrosis inflammation, poor regenerative responses, and we are able to show that old muscle for all practical purposes is to listen to young muscle, we're fibrosis and inflammation are decreased. And tissue repair after injury is not effective. Moreover, by the old tissue stem cells called satellite cells, and then for endoderm VRC, typically liver will also get fibrosis, adiposity and poor generation, and our experimental systems originate old liver turning it into young like, with fibrosis, the fibrosis and adiposity reduced, and the pathogenesis being increased. And then for up to dorm, the look at the brain, which is shared by many of our colleagues, everybody's interested in rejuvenation of the brain and cognition. Old brain is characterized by poor neurogenesis or formation of new neurons, increased neural inflammation, and also that leads to lack of cognitive capacity and memory. And be sure that our approach is now rejuvenate or brain making significantly younger such as the neurogenesis is improved. neuro inflammation is diminished, and cognitive capacity is enhanced. And I put 456, etc, because you pretty much now analyzed multiple hallmarks of aging and rejuvenation. So the axiom is that when several key organs become significantly younger than the entire animal will be also younger and healthier, and it will live longer. And I'm bringing this up because many people are asking, are asking me, did you do lifespan status or house further studies? And even though I will actually answer it, and we are on that path, you should realize that we are just the organ system composition. So if your muscle liver, brain bone immune system have become younger and healthier, by default, that means that you yourself are younger and healthier. And if you are younger and healthier, or the whole decline is no diminished, and how spawn is increased, you will live longer, there is no other way around it. Um, and then the opposite is not actually always true. So here is this image of very old supermodels. And living it is very old and sick, it is still alive. So it will be in a positive column. If people are just looking at the lifespan extension. So many lives, but extended animals are quite miserable.

Um, so actually, I have to apologize because I opened accidentally a wrong talk which is slightly outdated. So your permission Allison, can I exit from this sharing and open the more Yeah, thank you The pair.

In the meantime, I will let some more people from the waiting room in and since I can you tell us what was the outdated talk? When was that? Like?

That was a couple of months ago and and now we actually have additional publication that I wanted to demonstrate here. So just please bear with me.

I'm going to share in the meanwhile, a few ways in which you can plug in into the

Yes. Okay, just to make sure that I have a doc here. Yes, this is it. Sorry. So just that 2021 tokens that have 2020 and yeah, so let me return to zoom. And I'm

will go seamlessly from the previous slide. All right. I'm sorry guys about that. So it is because you know, we are all overloaded with zooming in with so many remote talks. And to have, you know, I'm happy to report that we are publishing every couple of months on this discoveries. And so I just wanted to give you the most updated version. So this showed quite a while ago that aging is malleable for liver muscle brain in the first study that we published by 2005. And also for neuroplasticity, cognition, spinal cord rejuvenation, kidney, bone, cartilage, etc. And those additional Business Awards were continuation of our initial research on heterochronic parabiosis by our colleagues. So this is the final slide, which is the Halloween party at bark Research Institute in Novato. And this slide is by Eric Burdon, who is the director of that research institute, which illustrates heterochronic parabiosis, we're an old mouse is physically sutured the young mouse, so they share our blood then they also share multiple organ systems. And while many people interpreted this as just blood shedding, you should realize that now this old mouse has access to young heart and lungs, and therefore just the oxygenation of organs will be better. And the old miles now has butter giant trucks, so it will absorb nutrients better, it will go to food and water more frequently and exercise because it has to run around with the young guy. So there are numerous numerous changes which are unrelated to the secret sauce and blood. And to make the system better controlled, we develop this small animal blood exchange procedure and apparatus where we exchange only blood, so the mice are not physically connected to each other. And this procedure takes about half an hour, and then they recover and we could see what happens to their organs and tissues are the younger or older. Moreover, of course, this procedure is not FDA approved, cannot be translated to clinic. But heterochronic apheresis is FDA approved, and it's called plasmapheresis. For people. It was not previously used for aging and rejuvenation, but rather for blood diseases, and autoimmune disorders.

So once you published on what happens with chronic parabiosis, and what is simply the cultural current blood exchange, typically our publications that are highlighted in the economist and other popular press, and everybody interpreted all this in the same way that young blood is a medicine. And here is the an army of people who tried to steal this young blood. But that is actually not the case. And so to give you an example of why that is not the case, I'll show you one outcome of heterochronic parabiosis, where young and old animals are physically sutures with each other, and blood exchange will the only exchange blood. So what you see here on green dots, and the red dots are the new loop forming neurons or neurogenesis. And that particular part of the brain and the hippocampus, which is called dentate gyrus, and it looks like this V region with many blue cells. So the edge of the Wii U, there are neural stem cells, which continuously make neurons even when mammals are already adults. And there is dramatic decline in this neurogenesis in the old animals, there is only maybe less than 25 neurons are made as compared to young animals, which means more than 1000s of them in this specific class. And then after parabiosis now all the animals made 50 neurons instead of 25. And young animals have a diminished capacity to make neurons when they are sutured with old mines. So we discovered this back in 2004, and shared our unpublished data with Coronavirus curry laboratory. And then they pretty much republished the same result Six years later, in 2011. So initially, we wanted to include the brain status into our paper in 2005 nature on muscle and liver and brain. But we could not include them because we had too many suggestions to do additional experiments in order to have my new job with DARPA. But we are happy that Tony's lab picked up on those discoveries and confirmed and republished but now if you what happens when you change the blood and animals are no sutures. You have the same dots here and the same v region can see them very clearly. But now even though the young animals have better neurogenesis than old, all blood dramatically inhibits neurogenesis dramatically. But young blood, all the young does not rejuvenate. So if you secure young minds, the dogmas and they have environmental enrichment and more exercise, their brain becomes younger. But if you exchange only blood, it does not become younger, which is quite interesting. And then if you look at their cognition or short term memory in this interesting test, which is called for limb hanging test, so mines are suspended upside down from the marsh wire, and they have to have coordination and grab to the wire, and then they don't want to drop down. So they have to remember that this is not good for them to just let go, they have to keep grabbing the wire. So in this test, which is similar for us to learn how to walk on a tightrope, for example, if we are learning how to walk on a tightrope, and this is initially there is not much difference between young mice and mice, because it is not a brute strength test. And after the young minds or Association control experiment with young blood, three out of four loan that they have to hang tight where, but if they are exchanged with old blood, now, three out of four do not remember or they cannot do it. And old minds, weather directions with young blood or old blood aren't capable of improving either agility and coordination or their memory. So basically just keep dropping now. So to summarize, basically, everything before neutral blood exchange. In 2008, Mike and I had an idea that multiple tissue in all mammals can be systemic originated an agent, and to publish the proof of that nature 2005. And the discovery was not about young blood, but the proof of principle that mammalian aging is systemic, right, that's in the blog, and it is reversible. So the question to us still remain, how did it happen in bloodshed and what happened? Meanwhile, this is my illustration of an intuitive jump because cats like jam, some other groups made this intuitive jump that young blood is the medicine and they pursued this direction. But we continuously continuously pursue, we were continuously pursuing the opposite directions such as that.

And this was not the opposite direction, it was simply direction. Why did parabiosis work in the first place, and if you think about it, there really is like a fork in the road. It could have worked because of the Youngblood or it could have worked because now we removed, antagonized or diluted, he elevated negative factors. So when young animal was shedding blood with an old animal, now there is less of those he elevated proteins, not only they are physically diluted, but they are removed by the young liver and young kidneys, for example. And then young animal can compensate also unchanged gene expression of antagonistic proteins. So there are numerous numerous directions that could be worth investigating. And so then what we discovered is that parabolas work because the old blood volution a young blood is not a medicine. And in fact, people who are pursuing Youngblood as a medicine avenues now run into a sort of the scientific wall, because young blood is not working. So now they're switching to exercise induced molecules comparing everything and the young versus old, are thinking about combination of Aden young factors, removing the old or another word to some degree, try to repeat the scientific and IP steps that you have already accomplished. And if you are interested, that can tell you more about it. So this brings me to the most recent discoveries. The main point is that the effect rejuvenation takes place without a putative cause, which is young blood. In the window, young blood definitely change with aging, as well as many other things change with aging. This is interestingly not causal for us for either aging or divination. So decline of young factors and young proteins happens but it does not cause aging and cannot cause their civilization. That is a quite interesting, scientific conclusion. Because many asked, many, many people asked me continuously to ask by Mao It doesn't mean that donating blood would make you young. It is a novel scientific threat for unraveling the costs of age to impose health decline, and then you meaningful medicine. So it's a big scientific direction and doesn't translate directly that if you drink lots of water, and then use the bathroom very often, or if you donate blood, then you become younger. So this is the paper that he published in May and then I will end up with the paper that he published in November of 2020. So the first paper will show that we can rejuvenate all three germ layer tissues just by changing our plasmid ceiling album without any young blood or non blood products. And so this is our small animal blood exchange setup, where we exchange young or old mice with the tube of over psyllium. So to water with 5% album man, and there is also blood cells because once we remove 50% of the old plasma, we then remove album and without which we cannot leave. And they also remove cells without which we cannot live. So they breed too much then return albumin and cells back. But instead of plasma with the circulating proteins, we now have source of water. And then there's a control for procedure because we use heparin and we use some surgical procedure we use young young exchange in old old exchange where young minds are exchanged with young blood, old mice are exchanged with old blood. As I mentioned, this is miniaturization of the FDA approved procedure for people, which has very similar if not identical principle, and it's called plasma for instance, or therapeutic plasma exchange. What we discovered is that oh plasma dilution is safer and more robust for rejuvenation than all other current approaches that not only young blood is not needed, but then when you perform this procedure, all tissues become statistically the same as you can sit down statistically different from you.

And then on top, now there is some examples. So on top you see what happens with mouse muscle. So this is mouse procedure. On the bottom, you see what happens the people clinical study. So with models muscle, the fibrosis of muscle shown here in this white areas, is diminished by one procedure of neutral blood exchange, and we call it neutral blood exchange, because our solution does not have age. And then the regeneration Are these the dentro of newly formed muscle fibers, which are different color, and the half central blue dots, which is their nuclear generation is improved. So this is a typical for younger generation, generation much worse. And now when young mice are exchanged with neutral blood exchange, their generation is dead similar to young and old mice exchange with neutral blood exchange now becomes similar to young as well and very different from old. And the same is true for fibrosis. So it is not simply that fibrosis is decreased in older animals after one single procedure of normal blood exchange, but that now by that parameter, they are not statistically different from the young cohort. And then they can also measure the diameter of the newly formed muscle fibers. And you see how they are not only denser, but they're also bigger, and which is also quantified here. So by my new parameters, muscle regeneration is now like, like young animals that are similar to 75 to eight year old people. And then what we did now, though, with Dr. Deborah Cooper, who is our key collaborator on the studies, we obtained blood serum from patients before threat to the plasma exchange versus after therapeutic plasma exchange. This is post therapeutic plasma extensions right after the procedure. And typically blood serum of older individuals is inhibitory for stem cells, which you might need to know because it explains why degeneration declines and why it is through blood. So here you see this one lonely yellow.as compared to many of these yellow dots on the right, and this yellow.is one muscle stem cell that is cultured and dividing in old human blood serum and there are many many cells here so all of the cells that are cultured are shown in blue, but out the wall of the Salesforce one is attempting to divide here and then non are dividing in the next in the next Part of the fever so the cells are cultured that don't serum then have decline in the regenerative capacity or serum inhibits their ability to divide. But after therapy, the plasma exchange disinhibition is dramatically less. So now many, many yellow dots mean that now cells are effectively dividing even though they are cultured with serum from exactly the same old person, but after therapeutic plasma exchange procedure, and they show that the theory is unrelated to presence or absence of human serum albumin, so whether we adopt boomin to culture so denote that delving into cultures, there is no facts if you don't think that albumin is the main factor in our experimental system. And then switching gears, the Olson demonstrated improved neurogenesis now you know what to look for, those are adults at the village and hippocampus. So now more neurons, many more neurons are formed in the old brains after this procedure. And here is liver, adipose, it's all of these red dots here are oil droplets or fat and liver. And there is tons of it in control animals, as well as tons of fibrosis shown here on blue outlines, and then both the adiposity is diminished. And fibrosis is diminished by one single procedure. So liver is also out there.

And then now, because many people were studying Young Blood effects on cognition, not just on the neurogenesis, but cognition, V, and then more recently published a paper on that as well where the procedure is exactly the same as I told you. But instead of sacrificing the mice and testing their various tissues, we look at their ability to remember and recognize new texture as compared to old texture and new objects as compared to old object. And you probably know that old people similarly to old animals are not very curious and inquisitive. So they are not really interested to investigate their environments, and they do not really remember what is new and what is all very well. So by this parameter by this whole multi parameters, single procedure of natural blood exchange, rejuvenated cognition. So now again, as you can see, old animals are no different from young animals and are very different from control animals that did not have this procedure. And then those of us wanted to compare the effects of neutral blood exchange with senolytics because this is another main profound approach to anti aging. And so first of all senolytics work very well. So this is 182 63. And senescence cells are shown here in blue by sa beta gal. So there are lots of senescence and old brains. And then senolytic reduces in essence, all brands were dramatically you can see here, but neutral blood exchange also reduces senescence the whole brain. So you can see here, so neutral blood exchange has senolytic or cinnamon properties. So, so you, in theory, do not need to use analytics if you use natural products. And then we looked at neuro inflammation. This is the last piece of data that I'm going to show neuroinflammation here as is illustrated by this red dots in the brain, which label microglia cells. So microglia are known to be like macrophages in the brain. And when they express this marker, cd 60 that means that they are activated and there is no inflammation in the old brain. And then little blood exchange dramatically reduces neuro inflammation. And that is probably why almost become rapidly more cognitively capable because we reduce that neuro inflammation. And then in contrast, if you look at seasonality, seasonality does not reduce more inflammation, which is quite interesting. So it diminishes load of senescence cells in the brain, but it does not introduce more inflammation, it significantly decreases how big these red dots are. But still there are many cells with those red dots which indicate neural inflammation. So okay, so then how does it all work? mechanistically and the way that it works is that aging is driven by access of sustaining proteins which are inhibitory for tissue health and repair. So we cannot the wolf out of them because at the young levels, they are really indispensable. We cannot live without them. But with age they become excessive and it is that excessive That then inhibits health and regeneration of multiple organ systems. consequentially, when we calibrate them, then you become healthier. But what is interesting and this is the proteomics data, this is the proteomics data on blood before versus after the procedure, this is mice, and this is people, what happens to the blood proteome after the dilute h elevated proteins. And what you can see here that there is lots of red on the right part, lots of red here, lots of red here. And if you look at the scale, red means that many proteins become elevated. which is surprising, right, because we diluted the protein, how can now we have all of these proteins which are now present at higher level after the procedure. And what you need to realize that this is one week to one month after the procedure is not right after. And the reason that this protein become elevated is a good reason is because those are the age diminished youthful protein, which we did not realize it didn't go anywhere, those are still encoded in our genome right now jeans, but they were physically suppressed by h elevated circulating factors. So once we diluted them, the repression is now also diminished. And we automatically restore the youthful factors as well.

And that produces the second wave of rejuvenation, the first wave comes simply from normalizing the level of H elevated proteins. But then once the levels are normalized, they cannot repress the youthful produce, which comes back. And that produces the second punch for improve tissue maintenance and repair. And they show the same specifically for brain proteins. So this is before procedure in mice after procedure in mice, these groups cluster differently from each other. And the same is true for people before versus after procedure. And these are the identities of the proteins, which then came back and are now elevated, and to have a table and our publication, published geroscience in November 2020, on all of these protests, and what do they do for the brain, they all are neuroprotective, and they all improve health of the brain. So I don't know I can stop here. Or I can then tell you why you know that epigenetics also become rejuvenated. And that as well as metabolism, mitochondria. And many other attributes and hallmarks of aging also become younger. But perhaps you could do it in q&a, just to give more time for Guinea. Allison, you just tell me, what's your preference?

Well, we do have four or five questions already. And I would just say if anyone in the audience wants to know a bit more about those factors, let us know. And maybe at the end of it, you can.

Yeah, because that it's kind of more educational points. It is not really about our discovery. Okay. So this is just our you know, this is the love and where you could really are socialized and did not have to distance. This is the behind our house beautiful heels. And this is Mike Conway, who runs love with me and our dog. This is me. We have excellent lab. We have phase two clinical trials underway with Dr. Cooper off as the clinical director and we do the science. The main conclusion is that large dilution of systemically loose TB resets molecular cellular and tissue determinants of organs to halt youth. Um, and basically that's what just on one or two parameters, but on all of the organ systems. Now I have I'm on scientific advisory board and have stock options with juveniles. And I hope to launch my company. I am you. So thank you guys, for you. Oh, yeah. Last but not least, we are very fortunate to be well funded laboratory. And these are the funding sources. Thank you guys so much for your, for your attention, and I'll be happy to take questions.

Well, thank you so much for being willing to speak. We have five questions already lined up, and I'm sure there's a lot more coming. Just tell us when you have to go. I'm just going to start right away with Raphael. Please unmute yourself and perhaps you can say one sentence about where you're coming from just to give some perspective.

Okay, hello, how are we now? Thank you very much for the presentation. You have to say I've been fascinated by it. By your research since I was aware of it and I've researched it over quite a bit. So I have I have a question. First question about it is what do you think the limits of it are? Because when you present it, it's quite, it seems quite comprehensive. So what do you expect the limits of the procedure to be?

Well, on this great question, right. So right now we have our clinical studies. And the main point of the study is there are two main points. One is that how long does one procedure last, say to have a positive effect? Right, then do we need to do another procedure a couple of weeks from Dan? Or can we do it every three months or every six months? So, what is the duration of the posting for that, since initial post effects are was quite large, then we can reasonably see the decay curve and we can really have this conclusion. And the second one, how many age associated diseases can we tackle because official trappy plasma exchange, which is FDA approved is not approved for diseases of aging. It is approved for autoimmune diseases, for example, multiple sclerosis, or rejection of transplanted organs. So for any physician to be able now to advertise and apply this procedure broadly for Alzheimer's or Parkinson's, or still persists and so forth, we need to do the study. So that is the second part is that what hallmarks of aging can be offset. And by doing this Raphael, we can then know the limitations. Because right now, we tested virus virus things in mice, and we tested effect of human serum on cultured stem cells. So then the limitations will be known after we do the clinical study.

Okay, thank you. Reena.

Do you mind if I stop and share your screen? I know that

I know what exactly I've noticed.

You want to share it again? Okay, let's say your next.

Yeah. Thank you for your talk. That was very interesting. I'm really big fan of your work. So I was interested in in in the mechanism through which book exchange is leading the rejuvenation, for example, in this case with senescence cells, is it that there is improved immune clearance of the cells and cells? Or is it that the senescence cells themselves are being dishonest, so to speak, back to normal? Thank you.

You guys are asking very insightful questions, by the way. Yes, I believe, or we believe that they become this in essence or those that are senescence and we still are investigating what are senescence cells, this is one of the challenges that they do not have any marker that will not be shared by normal cells that are not senescent. So P 60. And will be shared by cells which are known senescent. And even as a beta gal, which is the senescence cell marker could disappear when cells, for example, are split and are not overcrowded in the tissue culture dish. So what you hope is that, by normalizing quickly, acutely systemic protein will make many senescence cells less than us and throughout the body. And that's why we see less of them. So it does not work as senolytic, for example, senolytics just inhibit prosurvival molecular pathways in the cell. And therefore senescence cells are dying from that because they're damaged cells. We believe that we simply reverse their senescence is certainly yet to be determined. That is the hypothesis.

Lovely. All right. Next one up we have Robert.

Hi. Yeah. Thanks for that very interesting presentation. I just had a quick question of when you mentioned that the old blood

makes it worse for the young mice. How much worse how quick, how quickly did they actually get worse with your blood?

For liver and for brain they get quickly, very quickly worse, like in one week, and after one exchange, so which is quite surprising, right? They have 50% of the young blood still and they have all of the young guns so long to me yours no DNA damage, but simple elevation of age, no age excessive proton dominantly inhibit neurogenesis, cognition, and liver regeneration and these animals does not have negative effects on everything. So muscle is still young. But for select organs, the effects are rabid and robust.

So the younger animals are resistant to some extent

In the muscle there, Uh huh. Note in their brain, they are very non resistant. And then if they are injured, so if muscle was injured, then brain becomes old, much more robustly so and you know, it's kind of more fragile. And actually, I've heard from neuroscience, neuroscientist physicians that they observed the same thing that if somebody has any sort of peripheral injury and surgery, then there is negative consequences for the brain as well. And it could be because then there is additional inflammation and load on liver and kidney. And then brain basically suffers from that.

Okay, thanks.

Thank you, Joshua, Unix,

and iryna. Thank you for the great presentation. Just a quick question. So how do you think about aging being driven by an excess of systemic proteins versus that excess being sort of a downstream consequence of other drivers just high level or as deep as you'd like to go?

Well, of course, access to saving proteins is, is caused by some in tropical mountain chronology, right? That is for sure. I don't know if that is, I'm looking at my husband or my sister, because sometimes he has good good points. That of course, in the end, the access of seven proteins initially is caused by damage to cells and tissues. We know that already, right. So when cells and tissues are damaged, then you have certain proteins that are elevated and then their presence systemically. But interestingly, the being driven by sustainable proteins means that let's say that one of your tissue is damaged, not all of your tissues are damaged simultaneously, but you have a weak link, for example, there is inflammation someplace in your liver. Now, because systemic protein, those proteins are systemic, even though produced by liver, they will propagate aging and senescence to other tissues, making them down wars with respect to regeneration and repair. And that is one of the conclusions of our most recent paper if you guys want to do that we should publish in neuroscience and 20 point. But the general concept of course, we are we did not discover there is a general concept of systemic propagation of aging and senescence. answer your question. And then you might ask yourself, why do cells become damaged? Right? So all this every question is a step back? Question, or every answer is the button or the question. So why do cells become damaged? Unfortunately, for us, they become damaged simply because we eat and breathe from oxidative phosphorylation or mitochondrial activity, that is the major major cause of damage that we cannot avoid.

And really small follow up? What makes these excessive factors then the ideal point to sort of intervene in the biological cascade? Why is this the point to attack? So to say, if you will?

Well, because you cannot stop eating and breathing, right? I mean,

well, you know, versus

running Oh, but I mean, versus intervening on, say, I mean Damage Removal, versus repairing the machine.

Right, that the option to remove all of the damage will stop eating and breathing, which you cannot do, since you continue to eat and breathe, you will accumulate damage. And the damage cannot really remove the writing at every single level, every enzyme and cannibalize every organelle, every hole in every membrane part of every cell at is neither feasible nor necessary. Because his plant, he also developed the ability to repair and regenerate. And that is the capacity that is untapped. And what visual is that? With a very simple already of the approved procedure, we can make old organs and tissues identical to young or non distinguishable statistically from them.

Awesome. Thank you so much.

All right, love the next one up.

I mean, that was actually that answer was a perfect segue into what I wanted to ask because my lab is actually working on replacing and removing all the damage that occurs specifically in the brain using cell replacement strategies, so sort of a form of replacement.

Yeah, sort of a form of regeneration that you mentioned. And so my Question is in looking at, you know, the state of the brain after your treatments, you look at inflammation, increasing levels of neurogenesis, which are restricted to very small parts, especially in the human brain. And also cognition, right. So. So there are a lot of factors in the literature that have been shown to, to ameliorate these things and make them look more younger, right, so we even published a paper, you know, genetically manipulated one pathway showing that we reverse aging associated decline in neurogenesis with just one pathway. And that same pathway reduces inflammation. But when we look at the, these brains, right under two photon microscope, so the live brains, it's amazing what the difference is between a young brain and the old brain. So you can you can look in a young brain pretty deeply now with two photon microscopy, but by the time they're four months, nevermind two years, right, it's already getting harder and harder. And so despite an increase in the regenesis, decrease in inflammation and increase in cognitive function, this, this is, you know, this opaqueness of the brain as a measure of protein damage, and protein aggregates is still going on, on altered, right. So have you saw that? Yeah,

totally. First of all, thank you so much. This is these are very, very good points on the cell replacement, and gene therapy, those are their own, you know, interesting technologies with their own challenges, merits and demerits. So I will wait until gene therapy is effective, and it's much more. And the same goes for cell transplantation. And it is very difficult for the brain on numerous levels, or particularly considering that brain connections are who they are. And if you are trying to transplant cells, which established novel connections, that brings about the whole interesting set of problems and questions, will we be ourselves even if it works with scalability? And how now to cleanse and dishonor part of questions, excellent questions. And we just have recently developed a collaboration with UC Santa Cruz and Stanford to look at virus virus other things under two photon microscopy and brain sections. Exactly. Exactly. You know, what you said? Once you have your neuroplasticity things and synapses and bunch of other things that I'm not going to go into much detail. But to see to what degree did we rejuvenate the brain? Now to say that after one procedural brain will become young, I think it's not realistic. But to say that numerous parameters, little bit of genetics and gradually, all brains have become younger is what is our hypothesis postulates. And here, I bring this idea of the two waves, right, the first wave was simply to reduce size neuroinflammatory proteins, other age excessive proteins, then young proteomes, come back, and does its own thing and gradually improves your pattern regeneration. And then if you think about it, there are like 80 kilograms of tissue that will not become instantaneously younger. But if it becomes gradually younger, or does not become older, at the same rate throat, I think that is already a very good first output of the technology.

All right, thank you, john. I'm gonna ask this question just because I think he has some noise in the background. But he says, having read many of your great publications, it seems like there are a relatively small number of critical negative factors. Maybe it doesn't, despite the fact that hundreds of factors may be involved. Do you see additional advantages of depletion negative factors, specifically with affinity column versus dilution of all factors?

Yeah, so let's see, all of these questions from the audience today is just incredibly insightful. And I wonder if you guys are experts in the field, and I somehow missed it. And I would certainly look forward if you would like to send me emails and we can discuss it in more detail. But with respect to hundreds of factors, most versus few of them. Typically, there are certain heirarchy or dominance of interactions of key factors or determinants, and they control key nodal points. interactions of signal transduction networks. So really how cell behavior in health and repair is controlled is not by any one factor, but it is not controlled by hundreds or 1000s of them. It is controlled by few determinant factors which down when present absent for particular time and particular levels, then determine all of the networks. So, if you then identify those key factors, and you normalize them to help them use, they will take care of other hundreds of them. And this is again, not our discovery. It has been published over many decades on PubMed. In fact, my favorite thing is that if either one of us knew everything that was published on PubMed, you would just discover everything else. But, but based on our background and expertise, yeah, there are determinants, which then control networks have signal transduction pathways with hundreds of the factors. And you don't need to normalize or hundreds.

Alright, I'm going to not and Louis said, I have a question. I'm sticking in there. And also giving other people the space ask perhaps one or two more other questions. So when I asked you as I do with everyone, what is your key challenge that you'd like our accelerator projects to solve? This year? You just said safely calibrating. Secretary, will you to help us? Could you maybe elaborate a little bit more on that just in case someone was willing to tackle that?

Yeah, so that pretty much you know, I'm obviously our current interest in calibrating circle Tourmaline. We believe that the way forward is through removal like donation and dilution of elevated proteins. But certainly, it is just the very beginning amongst them that there's a scientific threat to unravel why we age and how to age slower or automate the process. So at some point, we have excellent screen systems and excellent computational biology, computational mathematical modeling systems, to identify every single determinant at elevated protein, and to see how to calibrate them to precisely young levels, not just in general, but individually, because we do not age identical to each other. So even though they will be the same patterns in our aging, they will also deviations from those patterns. And so, in my opinion, the safe calibration will be precision based or patient based, where for every person, we will identify your key proteome, the levels in composition, how does it differ from the healthy young proteome? And how can we reset that now to healthy young protein? Starting that is a big challenge, which will accomplish will provide meaningful extension of over healthspan and lifespan.

That's pretty succinct answer. I think there's a good challenge probably put in this book yourself. Okay, we have a few other questions. Let's see how many we can get through your very quickly we know

that this is because the audience is so well informed.

Okay. Yeah, I think so. Well, okay, let's, let's see if we can keep it up. Okay, linear next, and perhaps a few words about yourself.

Okay. Hi, I'm Lynn Cox. I'm based in Oxford. And my lab works on senescence, and we're trying to suppress the sass because we think that that's probably one of the most damaging parts of senescence. So I was really intrigued by your proteomics data, because I was expecting that the cytokines and chemokines would be going down, and yet they're going up. So that that was sort of contrary to what I was thinking, and then it made me think, are you actually rejuvenating the immune system? Are you promoting better immunological health? And could that be one of the reasons why you're seeing a loss of senescence cells? So are you improving senescence, cell clearance? And then the final corollary of that is, do you know if your animals have better resistance to infectious diseases? So if you actually improve their overall immunity?

Thank you so much. This is great question. So first of all, not just animals, but people who right now are undergoing the clinical trial, which is efficacy clinical trial, have improved resistance to infectious diseases, which has, it is like really amazing output that are in our we plan to publish the outcomes of the trials in a couple of months. And this is one of them. And we kind of mentioned that anecdotally in the papers that we published, but that seems to hold true. And immune rejuvenation is a main main aspect of that, you know, just absolutely correctly. I did not think about it, but you are also completely right, that if immune response is now younger, then senescence cells could be cleared. So not only, you know, hypothesis being that they become less and less and and produce less, but some of them the most decrepit ones could also no more successful eliminated by the system. Okay,

now, that's really exciting. I mean, with COVID, you wouldn't be diluting everyone's blood.

I know, but, but you started actually before COVID. So these are the clinical befores, a bunch of patients started in 2018. And remember that, um, that was before COVID. And in many, many of those samples were on the shelf samples. So they were treated with rapidly plasma exchange for something else, and we just got access to the repository. But what then will go on is that even though they never had the flu shot, but none of them had flu. So that was, that was like the end our older individuals. And so now these are these trends seem to hold true.

Fantastic. Thank you.

So another point, though, it is it is our now some of these people, they did get COVID vaccination, and therefore we have to stop the clinical study, because we don't want to dilute the anti COVID anti spy antibodies. So our clinical study would be slightly slowed down by COVID. Because when they get COVID vaccinations of this Laden, they cannot be prescribed procedures.

All right, thank you. Well, let's see if we can fit two more in and we've next.

Hiring as Christine Peterson, one of the co founders of foresight, if I understand the procedure properly, it sounds fairly low risk. And I was wondering, is there anyone, any MDS offering this now, in fact, I heard a rumor that the MD you work with in San Francisco does offer it currently.

Yes. So they'll build it and keep it all has 25 years of practical experience with crappy plasma faces. And in fact, he I think pioneered the concept of rejuvenative plasma freezes and already made a few innovations into the procedure that make it better. So he is our clinical collaborator, and perceived Chief Medical Officer of a company that is trying to launch. Now in general, if you simply use it as prescribed and approved by FDA to treat autoimmune diseases, then there is not many risks, certainly many fewer risks, as compared to injecting yourself with somebody else's body fluids like young plasma, there is some risk though, because your cells are being taken out of your body, spun down in the machine, and then mixed with the ceiling and album and then returned back to you. So it is really a complex thing. It is not something that you know, any doctor could prescribe. So um, so I would say that that is there are much less than four other alternative procedures, but they could they mitigate it, the physicians who are doing the procedure know what they're doing, and have been doing it for a while. That would be mine.

Yeah. Great. Thanks. Thank you. Um, well, we have one question here from Alexandria, maybe you want to ask it too. And I just want to piggyback on there. If there's a thing that this group could particularly help with your research and maybe Alexander, you want to ask you a question to. Hi, Rena. Thank

you again, for the update. Awesome, awesome new data. And I was wondering, like, if you can tell us anything about like future plans? Like, what what are you up to in the company in

the university?

Anything that you can share with the group?

Yes. So in the university, we plan to wrap up the the scientific results of the current efficacy clinical trial, which is a small trial, let's say 25 people to 40 people and publish it. So people know you know, the weaknesses and strengths and limitations and promises of this approach for treating human aging, age associated diseases on a company part of basically because I am university professor, I was looking for a business person to spearhead the company and finally, I was reached by email and now we have XLR to business person who started a few multimillion dollar companies recent past. And so he is basically connecting us with potential investors and we give the introductory talks on slide that's presentations. The idea for the company would be to launch the company at the time when the results of urine clinical trials come out and to have funding for Larger phase three clinical trials simultaneously as the first step of the company launch. And then the phase three clinical trial, we need to have placebo group randomization. And right now I have to mention it. Thank you so much, Alexandra for bringing it up. Right now our clinical trials are longitudinal study, we do not have placebo group we just compare before. And then after one round of procedure, 234, and so forth. And then how long does it take for for the positive effects to decay. So you have longitudinal clinical trial comparison features, IRB approved on a national level, but the participants are paying for procedures and are paying lots of money. So that's how during clinical trial was funded, and that's how it was approved by IRB. So then at the launch, which you hope to cover in May, we would need to have investment large enough to support phase three clinical trial where participants do not pay, and then we'll have more people and then and then to have placebo control. And we still plan to have the service for a fee, branch of the company to continue this something that we have proven to work already. So that's like, in a nutshell, you know? Yeah. What

would be what could people in this group do that would majorly help?

Well, that's working and advice, particularly in commercial real would help a lot. So then, you know, instead of making our own mistakes, you guys can tell us where to open not to make them how to troubleshoot. And then of course, you know, the major how to be with the resources with the, with the resources for phase three clinical trial. Because that is the very first setting that is the priority to have longitudinal trial success, when people pay, they absolutely have to be, you know, to have good standards of research and medicine to say that now we have phase three clinical trials, which, after successful completion, we can approve this procedure by FDA, for new classes of diseases, advertise it broadly, and perhaps have health insurance involved and compensated some of the procedures. So yeah, so the sources and networking, that would be, that would be a huge help. Alright,

arena.

What what's the high level? It's great that you're trying to commercialize this. And it's great that since it's already a procedure in humans, that you can jump right to late stage trials. But what's the high level story on why you can commercialize it? What can you protect in terms of intellectual property? Like since it's already approved? How can your company gather the money for it if you fund the trial?

Great question. So it is approved for completely different class of diseases. So if a clinician just wants now to start making money over it, or anybody prescribing it for Alzheimer's, Parkinson's, osteoporosis or anti aging, then they will have problems with FDA, they will have FDA warnings. And, and the only way to go forward in fact is to perform clinical trial to show that the procedure is effective for those classes of diseases. Now, in doing our clinical trial, they already know that the procedure has to be modified with all the trappings of plasma exchange plus, so it is not the identical procedure to the one that has been approved, but it is repositioning of either FDA approved processes and medicines. So it is not really novel chemical composition. And that is the first step in our intellectual property protection is trade secret.

Do you think you can protect the combination of tweaks or not

it is really a trade secret protection. However, there is a very tempting possibility to the patent for new application of patent. I previously have the approved and of patents, procedures, medicine device, so forth. So that is the easiest thing to do. And then of course, of course, there are numerous avenues for changing the composition of exchange fluids, such as that chemical composition itself as a protected domain, the procedure last longer to encompass more hallmarks of aging reversal. So that is the third way so they really start by you know notoriety, they are inventors and they already have numerous tweaks which we only we know how to apply successfully. And then the second wave is replacing thing of button things. And then the third coincidental wave is now normal chemical composition of exchange field. And so that is within like the first A couple of years. And then of course, you know, sky's the limit, because we just discussed here that we can identify for each person, the precise combination and concentration of things to be removed and to be restored. And there are additional patterns on how to do it safely. Like 710 years. Did that listen?

Well, we do have three more questions, I'm just going to read them out to you, and you just decide if you want to do any of them, or none of them. Okay. And so, we have, um, what do you think about existing clinics in Europe, for plasmas races? Then? We have, do you have any idea of the medically awaited cut off of the molecules you think I needed to be removed? And how long do the benefits last? And so and so how often do you require treatment?

Right? So the same thing about existing clinics, some people who are in Europe, they are, in fact get in touch with us. And they tell us that well, we potentially can do this, but don't really know exactly how to do it, you have no idea how to apply to anti aging at all, because it is not completely benign procedure, would you guys affiliate with us. So you teach us provide service and support and we give your share of profits. So don't break your profit now is has developed excellent isolation plan. And there are at least four clinics in us and couple in abroad who are interested in affiliation. So they pay upfront fee, and then we pretty much train them and provide support them, the patients are actually our patients. So they are working on that. Then the respect to molecular weight, no, they will be protein. So all kinds of molecular weights, and they will be elevated. So you cannot simply have a poor of 22 adult and remove aged factors. And that's why this really was laughable and non serious to find a work in production that will make you younger, there is no silver bullet one fraction they have. There are combinations of proteins and the published on those as well. So sorry, if I forgot.

But thank you know, you're awesome. Well, thank you so so much. I think we can all just thank you

guys,

much appreciated your interest and even getting in touch with us by

Yeah, please let us know how we can help. Thank you so so much. I'm going to post a link to those of us who want to stay on longer and socialize in a different room. No worries about you, even though please feel free to drop off. Please let me know anything that I can follow up with the group with and vice versa. Thank you. Bye, bye. All right, everyone. For those of you who still want to continue chatting, then I just posted a link in the chat. And I see many of you in the gather room, it's best to join in via Chrome. You can also now join via Safari, but this is a space for us to just all kind of debrief on this meeting. And yeah, thank you all for joining. This was fantastic. And I'll see you next week next month for the next proper one, but I'm hoping to see many of you in the Hangout afterwards. See you in a second and bye bye