The Peter Attia Drive - #27 - David Sinclair, Ph.D.: Slowing aging – sirtuins, NAD, and the epigenetics of aging

Episode Date: November 5, 2018

In this episode, David A. Sinclair, Ph.D., a Professor in the Department of Genetics at Harvard Medical School and co-Director of the Paul F. Glenn Center for the Biological Mechanisms of Aging, provides insight into why we age and how to slow its effects based on his remarkable work on the role of sirtuins and NAD in health and diseases. He also presents the case that stabilizing the epigenetic landscape may be the linchpin in counteracting aging and disease.  We discuss: How and why David moved from Australia to Leonard Guarente’s lab at MIT [7:30]; Sirtuins and aging [15:00]; A series of experiments elucidating the mechanisms of sirtuins [20:45]; How are sirtuins activated? [25:30]; NAD and sirtuin activation [31:00]; Nicotinamide, sirtuin inhibition, andPNC1 [39:00]; Resveratrol [43:00]; The NIH/ITP studies on resveratrol [55:45]; Does David take any compounds for longevity? [1:00:15]; NAD precursors (NR, NMN) and pterostilbene [1:02:45]; Female fertility and NAD precursors [1:14:45]; A unifying theory of aging [1:20:30]; Waddington’s epigenetic landscape [1:23:00]; If David had unlimited resources, what is the experiment he would do? [1:28:25]; Testing combinations to extend lifespan [1:31:30]; What made David aware of his mortality at such a young age? [01:33:45]; What is David’s book going to cover? [01:37:15]; and More. Learn more at www.PeterAttiaMD.com Connect with Peter on Facebook | Twitter | Instagram.

Hey everyone, welcome to the Peter Atia Drive. I'm your host, Peter Atia. The drive is a result of my hunger for optimizing performance, health, longevity, critical thinking, along with a few other obsessions along the way. I've spent the last several years working with some of the most successful top performing individuals in the world, and this podcast is my attempt to synthesize what I've learned along the way to help you live a higher quality, more fulfilling life. If you enjoy this podcast, you can find more information on today's episode and other topics at peteratia-md.com.

Hi everyone, welcome to this week's edition of the Peter Atia Drive. I'm Peter Atia. This week, my guest is Professor David Sinclair. He's a professor in the department of genetics at Harvard Medical School, and he is the co-director of the Paul F. Glenn Center for the Biological Mechanisms of Aging. David Hales from Australia, that'll take you about 10 seconds into our interview to figure out. He did his PhD in molecular genetics

at the University of New South Wales in Sydney. And it was at that time that he met the man that would go on to become his mentor. We talk a lot about him and this is a guy Lenny who's actually come up on other podcasts so I won't get into him now. But nevertheless, David met Lenny and decided he wanted to come to

MIT to study with him, which is what he did. And David has gone on to become one of the pioneers in a particular field of aging that focuses on a class of molecules known as sirtuans. I'm sure many of you have heard of sirtuans, but you might not entirely be clear on what Sir Tuen's are. I won't obviously make any attempt to do that here in the intro because we spend so much time talking about that during this episode. The other thing that David is really quite famous for is his role in the discovery of a molecule or class of molecules that stimulate Sir Tuen's, and by the time you get through this episode, you will understand why that may be a desirable thing to do.

And among these, the most famous is one called risk veritrol. Of course, risk veritrol came to fame probably a little over a decade ago when it was noted, at least in one study, in one type of animal model, to promote a longevity phenotype. And of course, what really made it interesting, at least for the lay press, was that risk of a withdrawal is found in very low concentrations in the skin of grapes. And therefore, the logic went, hey, grapes make wine, wine contains a spare withdrawal, wine makes you live longer, hallelujah, the French paradox has been resolved. Slight spoiler alert, that's not true, but we get into the wise that might be the case. This episode, we only had, when I say only, meaning we had less than two hours, but like

has happened in other podcasts, including the one with Rhonda Patrick, after we closed the podcast, because I wanted to be respectful of David's time, he had to catch a flight to New York right after we spoke. This interview took place in Boston. We ended up talking for another 15 minutes, and unfortunately that 15 minutes was some of the most Intense detailed nuance discussions of it and I left thinking gosh I wish we had more time to talk so I suspect David and I will speak again David has a book coming out next year and I think that'll provide another great opportunity to sit back down with him in this

Upside we talk a lot about his postdoc at MIT He came out of a powerhouse lab that has produced other notable folks that, including folks we've already interviewed, like Matt Kabralin and folks we've talked about, like Brian Kennedy. We talk about his contribution to the understanding of Sir Tuenz, which was something that was coming out of Lenny's lab, and David really picked that up and ran with it.

What are these things? What do they do? What's their role in aging DNA-repair gene silencing? We go over all this stuff in detail. We talk a lot about NAD and its precursors, specifically something called NNN. So if you are listening to this and you are in the camp of trying to understand how to make heads or tails of NAD,

NR, NNN, I think this episode will be helpful. Tragically, it was not until 24 hours after we did this episode that I ran into George Vlasik on an airplane who mentioned to me a paper by Josh Rabinoetz in cell metabolism that came out about a month before we did this interview, that we will link to that I think makes a very compelling case for the futility of

orally administered versions of these precursors. That would have been a really interesting discussion to have with David because of his expertise in this. At some point, I'll probably want to interview Josh and we'll go into that in really detail because, again, unless you've been up in the Himalayas hunting yeti, you're probably aware that these are the hottest supplements out there. We talk a lot about NAD levels. What do these things mean?

Where does it go? Where is it produced? We, again, as I alluded to above, talk about risk rare at all and its potential in life extension. We talk about the connection then between Sir Tuen's and NAD. We talk about his rationale for what he does. So David is actually very open about, you know, kind of the stuff that he does personally, including the fact that he takes risk rare at trial and met for him and then himself.

We also go into the differences between some of these molecules and again, a little bit to his book. There's a lot of stuff in here. Again, sometimes it gets technical, sometimes it doesn't. The show notes, as always, will provide a lot of help. So if you have any questions about papers, we reference the likelihood that they're in the show notes is actually quite high. Also, if you haven't signed up for our weekly email list and you wouldn't mind getting an email from me once a week, I suggest you head on over to the website, pdratiamd.com and sign up. Again, my promise to you is to make

it as non-lame as possible. And finally, if you are enjoying these podcasts, please head on over to iTunes and write a review there and begin. I would hope that you liked the podcast and leave a positive review, but I guess in the spirit of getting as much feedback as possible. If you don't like it and you have something to say that we can do to make it better, that's probably not an unreasonable place to leave that kind of feedback. So with all that said, please welcome to my interview with Professor David Sinclair. Good afternoon, David.

Nice to be here with you. Thank you for making time today. I gathered only by the commotion in this very busy place wherein there is a lot going on here and making time to speak with somebody as inconsequential as I am, is a big ask, so I am incredibly grateful. I'm pleased to be on. We were introduced by a mutual friend who's no stranger to people listening to this

because I've had him on before, David Sabatini. And maybe a month ago, maybe a little more than that, I said, David, you know, I really want to know more about Sir Tunes and who do you recommend that I speak with? And he said, well, I think David Sinclair would be perfect. So he reached out to you and you very graciously agreed to speak. So again, I thank you and you very graciously agreed to speak. So again, I thank you for that.

And I'm really excited to talk about something that I know very little about as you will undoubtedly learn in the next hour or so. Well, yeah, that's kind of David. Yeah, as you know, and probably the listener though, as well, he's a bit of a superstar, so. It's kind of him. I'll have to, you know, send him a note for that.

But, honestly, it's really great to be on because I know you delve into the details of the science more than anyone I've heard and I'm excited to be able to share that with listener. Well, yeah, that's that's the part that's got me excited. So a lot of times I've listened to talks that you've given and I can tell your I don't want to use their dumbing it down, but you're, you know, you're speaking to an audience where you realize that if you go too far in the weeds, they're going to miss the point. And so I found myself watching talks you've given on YouTube going, I wish he would elaborate on that point and that point. Hey, maybe they're at it. I don't understand myself. Let's see.

So I think just for a bit of background, I know you did your undergrad, your PhD in Australia, and then somehow you wind up in Lenny's lab at MIT. How did that happen? I've been interested in aging since I was four, since I realized that everybody and everything around me is going to die. That's a pretty big shock for everybody. Most people forget about it because you just kind of function, thinking about it every day. I forgot about it until my teenage years, and I realized that with the technology that was coming online, these are the days of early PCR and gene sequencing, we used to call it genetic engineering, I

thought maybe we're just the last generation who's going to live a normal lifespan, a regular evolved lifespan, and our children, and our children, children forever are going to be able to benefit from these new technologies and dammit. This is not right. I've got grandparents, parents, friends, and about 5 billion people who could really be benefited right now. So at that point, I decided to seek out the best people in the world and see if I could go work with them.

I got a PhD in molecular biology in yeast genetics, and that was a great PhD. And a real turning point for me was this guy from MIT, who I knew about because he was also a yeast researcher, he was a legend, I'd read all these papers. Lenny Garenti came to Australia, and here I am, this young 20 something year old, having dinner with Lenny Garenti and my PhD supervisor from Australia.

And Lenny starts telling this story halfway through dinner about this new project that a guy called Nickanora, Australia, Co and Brian Kennedy had just started doing and it was to try and find genes that control aging in yeast cells. And I said, okay, I know yeast and I've always wanted to figure this out. Damn it. That's what I want to do. Save me a spot, Lenny. I'm coming to your lab and he went, yeah, yeah, yeah, whatever, as he does to most people who want to join his lab. So I wrote to him pretty soon after maybe six months, I was finishing up, can I come to your lab and he wrote back like he does to everybody. Sure, you can come. But then a millisecond later, I was disappointed because he said, you have to bring your own funding.

So that was no small task. And that's a story in itself, just briefly though, this is a lesson for anyone who's listening who thinks that getting to somewhere like this is easy. It's you have to be massively determined, you have to have grit. And I just wouldn't give up because there's nothing else I wanted to do in my life. I certainly was looking at patent law even, but that would, I think, I would have died. If I had done that as a career, I just don't have the attention span. Anyway, I actually found funding. It turns out I applied to the Helen Hay Whitney Foundation, which is a prestigious foundation here, and they wrote to me and said, you can't apply your a foreigner. And I said, well, this is the old days with early email, I think it might even be post. And they wrote back and they

said, well, you're a foreigner, we can't afford to fly you out to Boston. Okay, I'm pretty good for an interview. And I said, well, I'll sell my car and I'll pay for the ticket. As I understand it, I was the first foreigner who was allowed to interview and I flew here. I stayed in Nicanor, Austria, I chose Basement. It was very kind. And my interview was with a guy who everyone was telling me is a real tough guy, super smart. I'd never heard of him. Of course, now everybody in science knows of Doug Melton.

He's king of, well, endocrinology and stem cells at Harvard. And I was just this kid showing up for an interview. And there was a line of people outside his door waiting. And I was fifth in line. And I went in and Professor Milton said to me, David, tell me what you want to do and I had literally five minutes to impress this guy, one of the smartest people you'll ever meet. So you've sold your car, flown 22 hours for a five minute interview?

Yeah, that was basically it. And so I thought about it most of my adult life, what was aging about? And a lot of people in those days back in the late 80s, early 90s were talking about the evolution of aging genes, death genes. And I studied evolutionary biology myself and that didn't make any sense. You don't evolve death genes. And I'd never subscribed to the group theory of selection. So every person for themselves, selfish gene, loved or can stuff. And so my thought was, the only genes that I could understand could evolve that have

relevance to aging are longevity genes. And right about that time, there was the discovery of in sea elegance, the nematode warmer of the Daff to mutation from. Yeah, it's about to say this must have corresponded with Cynthia's work, right, around the same time. It's just happening, and Lenny at the same time had just discovered a mutation that they weren't sure what up was doing, but it turns out it led to the Sir Two in story. And so I said to Doug, I want to come to Lenny's lab and discover life genes, genes that

give life and longevity. And I guess he liked the idea of what I was saying, or certainly very passionate. It probably didn't hurt that I brought a bottle of red wine as a present. I just thought that's what you do when you come to people's place. Later on I learned that that's equivalent to abriving somebody, but I was just a young kid.

But yeah, that's what happened. I got the confidence. No force shadowing the research role there. Yeah, definitely it was a suspicious kind of a thing, but being able to come to Lenny's lab, I would have worked in Lenny's lab for free. I would have lived in the basement and done it. That's just how much I wanted to do this. So I came over, it was late 1994, early 1995, when things were just starting to take off trying to understand what these mutations were that Brian had found. It was Matt Kabilin there as well. Matt's also a close friend.

He was not there yet. So the lab has produced just a ridiculous amount of sort of prolific individuals in this anti-aging space. It was a golden age. You could definitely feel something special was going on in those days. But I arrived, I was the first postdoc to join to study aging specifically.

Was Brian a PhD student? Yeah, so Brian, I was this new guy on the block. Brian teached me how to do yeast aging and he taught me. But all the other postdocs, they're probably 18 to 20 postdocs there, big lab, all working on transcription regulation. And that was the sexy thing. And I was told by, and I don't think I've told anybody publicly this, that the postdocs were saying,

you're crazy to work on this aging project. Lenny's lost his mind. It's a house of cards. It's going to fall down. You can't study aging and yeast. All of these things. And within the first few weeks, I thought, maybe I've made a mistake. Maybe I was just fooled.

Maybe Lenny isn't that smart, as I thought, but I stuck with it. But I do remember at one point on the phone, Nelly crying to my mother saying, I think I made a really big mistake. And that stands out as a point where I could have easily just quit. But I stuck with it and thank goodness I did because Lenny was right. And Lenny is really the smart visionary that saw that this was the right way to go. So you sort of alluded to it earlier, Lenny was sort of onto something, right? There was basically a new pathway.

Now, if we go back in time to 94, what did we know? Well, we barely knew about tour. I mean, it was just being basically figured out that this thing that Michael Hall had figured out a year earlier, Saul is figuring out, David is figuring out, I mean, there's still this early triangulation of what's going on with with rap amycin. At the time they didn't even identify recall. I don't think we knew that there was mTORC1, mTORC2, and they were doing totally different things.

AMPK is pretty well understood, Metformans been around, but I don't think people really had a sense at the time that it was anything other than an antidiabetic drug. So talk me through this whole sir-to-in thing. That's a pretty broad question. Let me narrow it down a little bit. Talk me through what you were just about to allude to. What was Lenny onto when you showed up? Well genetics is a fabulous tool because you don't have to go in with any hypothesis.

The biology will tell you the answer. And the gene that had just been cloned by Brian turned out to be what's called surf 4. Now, surf 4 didn't turn out to be a mammalian-conserved gene. So, surf 4 has not been as exciting, but there was this partner of the surf 4 gene. It was called surf 2. And surf 2 and surf 3 and surf4 form this complex approach and stick together and control of all things gene silencing. That's what C4 is for. Silent information regulator in this case number two.

And that was very unexpected. In those days, if you think back, the cause of aging was thought to be DNA damage mutations for your radicals. So we were all expecting to find genes that controlled DNA repair or antioxidants. At that time, because I go back and I think, at that time, I was still in college studying math and engineering. I didn't know. I didn't take a biology course, so I can't think about it through the context of my own education. How well was it understood that there were introns and extraons and that much of the genome wasn't even coding, was that well understood at that point in time?

It was in yeast actually, they were ahead of anybody else in eukaryotes. We knew that there were introns, and not many introns in yeast anyway. But we hadn't seen a full genome, it was still bits and pieces, maybe 5, 10% was known. My PhD was sequencing three genes,

that's all it took. We, so the things were changing rapidly. This new thing with PCR, you could move tubes into hot tubs and amplify genes. It was all very exciting. But what we didn't have any clue was that why a silencing gene that controlled negatively controlled genes, other genes, why that would have anything to do with aging, it was totally bizarre. It led to a string of cell papers.

And one after another, every few months we were actually publishing something new and we had a science paper. And that was the gold rush of this discovery because it really turned on the lights in this cave of a whole new area of biology. And we're still trying to understand actually why those silencing proteins are relevant to aging. But what we do know for sure is that these same genes, these serotones in all life forms, whether they're from plants all the way to us, do play a protective role in responding to energy and nutrients just like the Ampecine Snipe Topophthwadu. So you alluded to seroton, was that first found in yeast? Yes, it was. It was known already, actually, Lenny, and we didn't discover CERT2, it was

already known as a science in protein that controlled the mating type. In other words, the sex of a yeast cell. And if you don't have the science in what happens is that the yeast cells get confused, because they're now turning on genes for A and alpha, which means male and female. And a yeast cell that doesn't know if it's male or female will not mate and it's become sterile. And turns out that's a hallmark of yeast aging, is sterility. So if you, you know, the way you can tell whether a yeast cell is truly old, is it sterile or is it just sick? And that's how we used to tell. But now we actually understand the cause of that sterility. It's the actual, the movement of Sir Tuprotein away from those genes that it should be at to go deal with other problems in the cell.

I see. So it's not that Sir Tuprotein becomes deactivated. It just, for lack of a better description, shifts its attention elsewhere. Right. It becomes distracted by other things going on in the cell. And we had a cell paper in 1999 with Kevin Mills and Lenny where we discovered and a couple of other groups should also get credit for code discovery and this is that the Sir-Two and they're also involved in DNA repair. When you get a broken chromosome it's the Sir-Two complex that goes along helps unwrap the DNA we think and put it back together and repair that and while the Sir-Two complex is doing that it cannot be also silencing.

There's not enough of it to go around. And you might ask, well, why would this all do that? Why don't you just make more so too? What we think is that there's a very ancient system that coordinates controlling mating and DNA repair. You don't want to be mating and dividing if you've got a broken chromosome. So this is a way of coordinating those two events. And it's very ancient, it's a very active system.

You need DNA checkpoint signaling, so it's not just random. But what we also have come to realize is that this, let's call it this distraction of the serotones, it's conserved. We find this happens in our own aging process as well. So there's really two roles. There's gene silencing and DNA repair. Now, sertoins are HDAQs, is that correct?

Are they all HDAQs? So HDAQ, histone deacetylases, this is the old name for protein deacetylases now. Because well, we've all come to realizes histones are just one of the things that sertoins and these are HX2. They can target what are called non-histone proteins and they remove not just a subtle groups but also other types of what are called generally asyl groups. So that's a whole new world. That means that serotones, the family, there are seven of them in mammals, five of them

in yeast, target other proteins. Proteins that are inside a plasm in the nucleus, even in the mitochondria. And that's their role. It's not so much only about controlling the chromatin and histone, but also about controlling signaling and metabolism as well. And they can do that by targeting

any protein theoretically in the cell. So what are the, you know, again, thinking back to DAF2, DAF16 as the parallels with Fox O and IGF, were there elegant experiments in the yeast that could show you extreme conditions of lots of cert, no cert, and what that phenotype is? Oh, yeah. These were the first experiments. I'll try to take it through them correctly in sequence. So what we showed with Brian, first of all,

since 1996 cell paper was that we were looking for this movement. There was this so-called age locusts. We didn't know what it was. We didn't know where they were going. We just know that they knew that they left the sound-mating type locusts. So what we did was we stained it. So Brian had moved on, actually, to his postdoc. And Kevin Mills and I, student at the time,

our job was to find where the sort of protein's going. So Brian had moved on actually to his postdoc and Kevin Mills and I, student at the time, our job was to find where the sort of protein's going. So we stained them and we could look at the month of microscope and what we saw was they were going to this little place in the nucleus, which we eventually figured out was the nucleus, which is what makes the RRNA, which makes the ribosomes, it's a really important part and the DNA that's within the nucleus is called the ribosomal DNA or the RRNA, which makes the ribosomes. It's a really important part, and the DNA that's within the nucleols

is called the ribosomal DNA, or the RDNA. And that's where they were going, not just during normal aging, but also during an accelerated form of aging. So there's a whole story that was lost in history, actually, that maybe I'll just quickly touch on. One of the first things I did when I got to Lenny's lab was to work on Warnes Syndrome, which is premature aging disease. And these are kids that die in their teens or 20s,

aren't they? No, that's a different Hutchinson Guildford Syndrome. This one Warners, they lived till 40s. Yeah. But the gene was just cloned by George Martin and his team out there. And the homolog in yeast is called SGS1. And I picked up the paper it was in Science I Recall. Went into Lenny's office and I said, I've just been scooped. But there's a yeast homolog, I'm gonna work on that. Is that okay? And he said, yeah, go for it.

And so we worked on SGS1 for a little bit. And what we found was that they were going through accelerated aging as well. So we had a science pair on that. And what was exciting about that was that they were also becoming sterile and the sort of complex was moving as well, just like the normal aging. So we had this model, rapid aging model. And you might say, well, so what's the big deal? You've got a rapid aging model. But A, that told us that there was some universal

process that results in premature aging in humans, probably in yeast, the same thing. And we could also study this much more easily than an old yeast cell. Consider that to find a single old yeast cell, it's really hard, especially if you're just studying replicative aging, which is the number of times they divide. Each mother cell produces, on average, 10 to the power of 25 offspring. Okay, so that's by my calculations, about 30 million offspring, and you have to pull out that one cell and study it biochemically. Wait, you said 10 to the 25. Yeah, that's my...

I'm sorry, 2 to the 25. Oh, okay, okay. Yeah. But in any case, that's way more... So what we used to do is to sort out the old cells. We'd label them with a chemical and pull them out with magnetic beads. But it was a real, we couldn't get many of them. You'd just get a handful.

But with these SGS-1's protein, you could get bunch of them. The mutants, we could get a lot more. And so we made a lot of progress using that. But every time we made a discovery with the SGS protein, mutants, we went back to the normal yeast cells and verified. But we were actually able to figure out what the distracting problem was for the sir complex. And that was actually DNA breaks

and DNA recombination that was occurring at the most repetitive regions, the most unstable region of the genome, which is the RDNA, which is in the neclealus. And that was what was distracting those proteins. Interesting. When you knock out sir two, is it easy to knock it out? Yes, that was the next experiment.

Okay. So then we wanted to know the prediction is if you knock out sir two, you should get a lot more of this instability at the Ardena. Yes, and the question is does it translate to accelerated aging or not necessarily solid aging, but more cancer or some other phenotype, right? Well, in yeast it led to accelerated aging through the process I was telling you about. You know how my constability, DNA repair went down. And also that happens in animals.

Although it's a little more complex because it's embryonic lethal in a lot of mice. So you can't easily do that experiment. But what you can do is the opposite. You can turn on or over express the serotonin in a yeast cell. And if we're right, you should get a few things that are going to happen. You'll have more genomic stability at this,

particularly at this RDNA locus in the Neucleolas. And the yeast cells should live longer. And that experiment was done by an incoming graduate student, Matt Kablein. And what a fantastic project to get when you walk in. He did it. And the day that he got life-plan extension with Extrasser II was a very good He did it and the day that he got life-band extension with Extrasser

2 was a very good one for him and the lab. So what's the next step from there? The most obvious thing is how do you develop a compound that would do this without the genetic mutation that empowered it? Yeah, that was the issue because you can't easily genetically manipulate humans. So the question was how do you turn on these genes? Now we we spent about three, four years working on caloric restriction in yeast and then in mammals. And my lab and some others were leading the charge and showing that Sir Tuan's both in yeast and mammals were not only necessary but was efficient when you overexpress them to mimic calorie restriction. Put

another way if you knock out Sir Tuan you don't get the sir-to, you don't get the benefits of calorie restriction. You don't get the benefits of CR. Right, and that's also now being shown by others to be true in mice as well. So that led to the idea of it. Is that true in all mice? I couldn't say, you know, there's, even in yeast you can get around the need for sir-to-ins

if you stress the yeast really intensely with very little amount of calories. But there are, you know, there are aspects of calorie restriction benefits such as stress the yeast really intensely with very little amount of calories. But there are aspects of calorie restriction benefits such as a lifespan extension that are ameliorated, lessened by a serotonin knockout, but it's still complexified by the fact that it's lethal in embryos. And you have to knock it out in the adult to do a really strong... Yeah, it's the same sort of issue that Cynthia had with the Daphs, which was if you do too

early, the C. elegans doesn't make it into an intermediate stage. You know what, somebody hasn't even done the proper experiment, which is to take a mouse that you can knock out sort one in an adult whole body and then calorie or strict, something that we probably should have done years ago. We didn't, but the technology is there to do that. But what we did learn actually was both in Eastern and mammals was that it's not just one of these genes that's important. It's the whole family. And that if you knock out, take

yeast, for example, if you knock out sort of two in yeast, you lose the ability to respond to some mild calorie restriction, but if you really calorie strict them, they'll still live longer. And there was a big debate actually between Brian, Matt, myself about that. And where we settled on was that these other search, search who related genes were also helping and they work as a family. And if you knock one out, the others can compensate. The yeast only have one. Yeast of five. You still five. And humans have a whole family like eight or something or seven.

Yeah. Okay, so only two more. That's interesting. Little know, in fact, most people ignore the other yeast or two ends, but they're just as interesting and what we've found is they also can extend lifespan as well. So when you, actually, it's funny, I was gonna go back to somebody else, you said a second ago, but...

What's the teleologic explanation for why caloric restriction and Sir Two Ends would move hand in hand. I'm just like from, you talked earlier about your appreciation for sort of evolutionary biology. So an organism is, you know, in a nutrient deprived environment, it still has to be able to do a bunch of things

if it's going to be fit. Do we believe that that's an environment where we need to see more stabilization of the genome or repair or silencing, or what do we think is the biggest insult during that period of time? Yeah, so the biggest insult to any life form is a broken chromosome. That's lethal if you don't fix it. And do we see that more likely happening during nutrient deprivation?

I would see that as I would have assumed, to be honest, I would have assumed that was independent of nutrient exposure or at least if anything inversely cooked. Well, if you don't have enough nucleotides to complete replication, you're going to break a lot of lines. I see. So they do go hand in hand, but the bigger picture is that the sartouins evolved. We believe what are we talking about? Three and a half billion years ago in the first life forms, early life forms, maybe just

off to the first one of the first proteins to actually evolve, we think would be a seroton. And its job is to sense stress, biological stress in the environment, whether it's DNA damage or it's a burst of cosmic rays, change in temperature, or a lack of nutrients. And their job is to allow that organism to hunker down and survive. Stop mating, stop breeding, we can do that in another day. If we don't survive this, our offspring in a dye anyway. So they control, we think they control which genes to turn on and off in response to adversity and they allow those organisms to survive. But they're also talking to other pathways, so they're going to talk to M2A, they're going to talk to Cynthia's Deaf pathway.

And collectively, these are the genes that we've settled on as the longevity pathways, but they didn't evolve for longevity, they evolved for survival during adversity. How conserved are these across, let's use the big four models of eukaryotes from yeast, worms, flies, larger mammals like mice and rodents. Is this relatively well-conserved the way the tore pathway is conserved or does it have more bends in the road? Well, they're surprisingly conserved that you can just manipulate one gene in each of these organisms and get lifespan extension or one drug, works in all of these organisms. I would challenge anyone to use a chemotherapy to help a yeast cell.

So this is quite a magical discovery that the same pathways are that well conserved and that ancient. And that's actually one of the advantages we have. Aging is actually not that difficult to be able to control, and that's because our models are very good. If we can, I truly believe that if we can extend the lifespan of a yeast, a worm, a mouse, humans are so close. Right. If you can do it across a billion years, you should be able to make that leap to a few other hundred million years.

Exactly. It's really just the regulatory agency's making sure that we don't do any harm and it's safe. But the biology is all still there going way back three and a half billion years ago. So it was 2006, 2007. When did Resvera Troll emerge as an early, sir, two, an activator? 2003. Okay. And the story behind that was that we were looking for an activator. We're hoping for an activator.

Are you in your own lab at this point? Yeah. You finished your postdoc I'm guessing. Yeah, I managed to move to Harvard in 1999. So that was the year when a lot of things happened. While I was moving, we published this dinner repair. But just going back to the silly sort of social question, did you at some point think I want to go back to Australia and set up a lab here?

Like was it a difficult decision for you to stay in Boston as opposed to? Because you came from Sydney, if I recall. Right. Well, the goal was to, because like those are pretty different climates. I've been to Sydney once. I spent two weeks there. I could stay there. Yeah, I miss that, but I also like adversity.

I thrive on adversity. And so I've come to it like living here. The intention was to come here for two years. I had a job to go back to. And, but the first week of being here in Boston was like a city I could only dream about. This is the Athens of Ancient Greece, Rome of Ancient Rome. For biology, this is it. So I was in heaven. I'd never experienced a city where you're on the train and people around you are reading Science magazine and Nature. That's my dream.

So it was a very easy decision not to go back to Australia for reasons that if you really want to change the world, you've got to do it from here. So now you're in the process of kick-starting your own lab which comes with its own stresses, right? You've got to secure funding and all those other things. And then what's happening on the front of this stuff? Yeah, so 99, a few major things happen. One was this DNA repair connection. The second one was

Lenny's lab published that NAD was a requirement for us to do an activity. And Shinemi, who's a wash, you know, was the post-doc who made that somewhat serendipitous, but brilliant discovery. And then the third thing was the connection to calorie restriction was happening around that time too. Going back to the second thing, we talk about it now today, like it's in textbooks, it's so obvious, right? That Sir Tuen's our NAD dependent DS satellites. Okay.

NAD is so ubiquitous in cells that if the quantity you could have in a cell varies on a scale from 1 to 10. What is sufficient to produce this DSLase activity? Is it anywhere from 2 to 10, or does it have to be quite a high concentration? It can be 2 to 10. You can actually get very low levels in some disease conditions, and the animal is still alive. So it's basically only saying that NAD is necessary for certain to-ins to work.

Right. Well, without NAD, we'd be dead in 30 seconds. But by other reasons as well. I mean, wouldn't we be dead just from not being able to do electron transports? We would. And there's more than 500 reactions that you made just to survive. But what we didn't know in the 2000s, and it was actually quite

crazy thing to think that NAD was regulating anything. And that was actually what we first worked on in my lab was the control of certain with NAD levels. The reason it's crazy is you read textbooks and NAD is that the most ubiquitous important molecule in the cell, how could it possibly be varied during aging, let alone during the day when you eat something or you're circadian rhythm? Now it's obvious, we know NAD goes up and down, it changes with age, but in those days, people thought if you changed NAD levels, you'd probably die. That's not true. Was it known at the time that the doesn't complex one of the mitochondria basically convert NADH to NAD. So you would at least know that in the mitochondria the concentration of NAD must go up and down or else you couldn't actually respire.

Well, locally it goes up and down, but the steady state level is pretty constant. And when you say going up and down, are you talking cytoplasmic, nuclear plasma, where are we talking about? You know, we had a paper I think was 2007 where we had a quite a surprising result which told us that it's not just the cytoplasm and then it goes up and down. It was also the mitochondria going up and down. And we didn't know that.

We actually stumbled upon it. We found that if we kept cells with high amounts of NAD, they would survive better DNA damage, others in salts. And we could deplete NAD in the cytoplasm, but the cells still lived. And we didn't understand that. How did you do that, by the way? How did you deplete NAD?

Actually, come to think of it, we could either over express NAD depleting enzymes, but actually the way we found it was that when you damage cells with the DNA damaging agents, say, chemotherapy, drug, the cells themselves to complete NAD naturally with this enzyme called PARP-1, which is an NAD consuming enzyme. And that's very well known that if you hit a cell with a DNA damage agent, the reason that it dies is NAD depletion. So we were measuring the NAD depletion in the context of...

And that's because the cell is trying to utilize that an AD to repair the damage you've just caused. Right, right. Or you didn't know that at the time. No, that was well known. So part one is a no-Indian air repair protein. And that's, if you block part, you also protect cells. But what was interesting was that we could over express, give more copies of a gene that made NAD. And this is called an MPT, which is the equivalent of the DNC1 gene in yeast, which we found was important for lifespan in those organisms. So we were overexpressing this NMPT.

Cells had more NAD. We hit them with the toxin. They'd survive better than regular cells. But the NAD was still being almost completely depleted from the side of it. Sorry, just because this is sort of at the crux of it. Did they survive because they were unable to deplete their NAD? No, we saw that NAD just disappeared from the cell, but they survived.

But they still survived. But there was a place we weren't looking at the time, and we tracked it down to the mitochondria. So this begs the term, the mitochondrial oasis hypothesis. So myself and Anthony Solve from Cornell coined this term because what we found was that as long as the mitochondria stayed active with their NAD it didn't matter the cell could survive and recover from that stress. And it actually turns out that the levels in mitochondria, NAD levels are even more important than cytoplasmic NAD levels for survival. That makes sense. And of course, that's a hard thing to measure, isn't it?

It was extremely hard. That's where Anthony came in. Anthony is a chemist and a biochemist at heart. And it was extremely hard to isolate these mitochondria preserved the NAD. And we had to use new technologies to be able to do that. Well, how was that done? We were using my spectrometry for the first time

to measure NAD. And what Anthony did brilliantly was to make labeled versions of NAD and its precursors and he could spike those in and use those as references to measure NAD levels in these compartments. An NAD, if I recall from just biochemistry, you don't get to move that in and out of plasma into cells. It's made de novo in the cell. You have what you have, You can make more. You can bring in precursors, but you don't get to shuttle NAD between cells, correct? As far as we know, right? Okay. Yeah. It's sort of like an ATP problem. It's really hard to quantify ATP in a cell. By the way, does NADP do any of this as well? Well, it's important. No doubt it's part of this whole problem that we're working on. But if you add NADP or even NADH, which is only different by 100, they don't, those

don't work to activate such a one. Only NAD plus will do that. And I mean, nothing, a two nerdy on this or is it, it's obviously more than just the charge, but the charge must play a role if NADH can't work, right? Yeah, so it's probably a combination of the charge and some other size. Yeah. Okay, so now you've made this these three discoveries, the second of which we just went into in a little bit more detail. How does the story unfold now? You've got your spanking new lab. Well, you worked for a little while on the SGS-1 Wernos protein, and Tila Mews put out a paper on that. And we didn't work on sort of two ends for about a year

because Lenny said, I don't want you working on sort of two ends when I left the lab, which was a bit of a shock. Why? He doesn't want them what the competition I suppose, but I thought a year was enough to give him a head start, but we quickly started working back on that. Is that a common request if people when postdocs leave their labs? No, of course not, but what am I going to do?

I mean, the guy trained me, I go in my career. But in those days, it was very competitive. So the Lenny of today is not the Lenny of previous years, and he's a wonderful friend and mentor to me. But that's how it was in those days. It was very cutthroat. I see. But if it were up to you, you would have continued to work on sertuins.

Right. So then now we're basically back to 2001-2002. The moratorium is up, you're now back to working on it. And nobody at this point in time has yet figured out a way to exogenously manipulate sertuins more than the stuff that we've already talked about, which is nutrients, stress, and things like that. Yeah, we were trying to feed yeast, NAD, and we gave them nicotinamide, which is a precursor to NAD, vitamin B3. And actually, Kevin Bitterman, who's now a very successful venture capitalist,

worked my first student. He put nicotinamide on yeast, and we could measure the sur two activity by the color. And if sur two was more active, they'd turn red. And he walked into my office, one of his first experiments, when he's worked on suruan, and he said, David, something's weird. We didn't get activation. We got inhibition. So the yeast had gone red. And I said, Kevin, it doesn't matter what happens. As for some expected, that's even better. So that led to a paper that said that vitamin B3, high doses is inhibitory of Sir Tuan. So now the labs around the world use nicotinamide as an inhibitor for serotonin. I wouldn't recommend taking really high doses of nicotinamide. Why is that?

There's a few answers. Bichemical answer is that there's an evolved pocket in the serotonin structure that measures nicotinamide levels and it's a feedback loop. So nicotinamide is to get nerdy is the product of the reaction. It takes an AD, cleaves it. Oh, I got it. So it's just a negative feedback loop. It's seeing too much B3 and it's saying, I have too much of my output. Start it down.

Start it. Turn it down. Exactly. So we struggled with that. We couldn't get NAD to go into cells as too big, even with mammalian cells as difficult. Now, so we were looking at ways to make more NAD in the cell. That was our original thesis before his Viratrol was on the radar. And we were turning on and discovering the genes that made NAD in yeast.

And we cloned some of the genes in that pathway. And there was one particular one that was called PNC1. And it had been studied in the context of tuberculosis. And what we found was that when we, calor, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, we, got to do with stress, but we knew exactly what was happening. This is a stress response that was turning on NAD, production, and activating Sir Tuan's. So we had our first nature paper actually on that 2002, I think, 2003. And we found that PNC1 could mimic caloric restriction and raise NAD availability. And then if we knocked out the PNC1 gene, yeast cells didn't live longer when we calorie restricted them.

And what's really interesting about that I think is that PNC1 doesn't just get turned on by colic restriction. It's turned on by heat, low amino acids, salt, high salt. And so this is a gene that senses the environment and turns on the serotones. Exactly what I was explaining earlier about those early life forms on the planet Since their environment and through NAD and other ways they can turn on these pathways for defense So where did you go from PNC1 to resveratrol? well, we

We teamed up with a company at the time they were called Biomole and they were making reagents for H.DacS's this kind of thing. And Conrad Howitz was a scientist there, and he had invented, wasn't even available yet, an assay for Sir Tuan activity in vitro. I forget why he reached out to me, but he said,

hey, well maybe I wrote to him, but whatever happened, he sent me some kits to test. And they worked, and it looked great. What was the gold standard prior to these kits? Oh gosh, I mentioned this to an activity required Western blotting, which is detecting the cell groups. It was horribly hard. Still is. And this was a very quick way to look for

molecules. And what Conrad did was to take that assay and look through his collection of molecules at Biomole, they had libraries of these things. And two of the first molecules that were discovered by him to change the activity of sort one, humans are one, were a couple of plant molecules. Now, he found plenty of inhibitors. That was, that was. Inhibiting it wasn't the hard part.

Exactly, but he called me up and he said, David, this is something's weird. We've got these molecules that seem to activate the enzyme. And I said, that's great. That's what we need. And he goes, well, I don't know if it's real, but let's work on this. So we worked on it together for about six months, trying to prove this wrong. Because in the history of pharmaceuticals, there's only been a handful of molecules ever that have been true allosteric activators, as these apparently were. But we thought it could be fake, it could be messing with the assay, it could be an antioxidant activity.

But what really clinched it for me at first was we put these molecules onto cells and they lived longer, but only when the surgery gene was present. And the chance of that happening and us being wrong about this idea were pretty low. So that was a good sign. But the other thing that's not well known in history is that those first two molecules that Conrad how it's discovered, they were not resveratrol. They were actually, I think, Pisiotanol and Chorsotan. Chorsotan's found in fruits and onions, I think. But there was a brilliant CEO, Rob Zipkin, and Rob was a chemist, and Conrad showed him these structures that were quite similar, these two

molecules. They look like two rings connected by a bridge. And Rob said, hey, you know what, these two molecules look like? Resveratrol. And so Conrad and I are like, oh, what's resveratrol? Anyway, I certainly was. I didn't know much about resveratrol, but I googled it and it comes up with, I was suspected ingredient for red wine and I could just see the next 10 years of media, which actually happened, but it was an interesting time because we were it was the first molecule that that I can recall was clearly lifespan extending in an organism that you had to find genetic pathway that wasn't just found off the shelf. We really knew the pathway and we figured out, and it's still as far as I know, the only

drug program that has come out of the aging field where it was by design that we find these molecules. So full credit to David and to others who have their molecules that have come out of drug development. So when they first started to make risk vera-trol for the purpose of these experiments, they weren't extracting it out of naturally occurring sources. It was just being synthesized straight away, even though it was acknowledged that hey, this thing also exists in nature. Well, no, I think that probably was an extract, but I'm not being very articulate, but the point I want to make is we did it in a different order.

So repamison and metforin were already on the shelf doing other things discovered by others for doing things, and then you can trace them back to the pathway and you get lifespan extension. We didn't do it that way. We said, this was the direct target. We went genetics first, and then found a molecule and then tested on the organism, which is, you can do it both ways, but this was more rational science. But anyway, that's the reason, but we also had this very interesting result that we found

a yeast mutant that, and actually a certain one, mammalian mutant, that was blocked, that couldn't respond to as a vector. So we already knew that there was an amino acid that was required for this activation by resverteral in the test tube, which told me this isn't just some random thing. This is a highly conserved mechanism because we could also show this amino acid was found in other organisms as well. And what amino acid was that? In sort one, it's called E230,

and the mutation made a lysine. So it went from a negative charge to a positive charge at that location. We didn't know why it blocked it. Now we do, but that was one of the early things that we had in our knowledge base to be able to go forward. Now, when you put respiratory in cell culture, like you had with the previous two compounds,

you mentioned with the previous compounds, if they lacked Sir Tuhin, they did not get the longevity phenotype. Did you see the same thing with the risk-veritrol? We did in yeast, and then in worms, and then in flies. So the fly work, a couple of Brown University scientists, Mark Tater and Stephen Helfand, did the fly work, we did the worm work predominantly, and that went into another science paper, I think 2005, 2004. And what was exciting about those days was that here we had things as distant as a yeast cell and a fly separated probably by 500 million years or more, and we had a sur two dependent drug lifespan extension, molecule lifespan extension, which to me is that we're on to something interesting here.

Yep. Remind me, what was the phenotype of the non-sert bearing organism? Oh, now you're stretching my memory. In flies, I don't recall there being a strong phenotype if you're asking where they really short lived. Yeah, yeah, yeah. That's what I'm basically trying to get at, is when you demonstrate the first proof of concept, which is this drug won't work on you

if you don't have CERD. In other words, I think where I'm really going with this is are there other plausible explanations for how these compounds could have extended life outside of CERD2N? Oh, yeah, yeah, sure. There's plenty of possibilities. I mean, it could have just been that the CERD2Ns

were making them so sick that they weren't. But one of the reasons that we thought that it was real was that we tried to rely on every other possibility. We fed flies into accidents. Even if you got life saying such a wasn't through the serotones, you can only make hypotheses and test them as best you can with the tools you have in those days. That's what we did.

But now we can actually go back and replace the SIR2 and mutant with this point mutation that only knocks out changes one amino acid, which is real, that's real micro surgery. And when we do that and we go back now, we can still see a requirement for SIR2 for these effects. So it was the mouse paper in 0506 that was the one that kind of brought this to everybody's attention, even outside of the aging community, correct? Right. Well, so the the residual story in yeast was one thing, but when the mouse paper came out in nature in 2006, it went global and it was caught the attention of probably most people on the planet. Certainly those who drank wine paid attention.

The wine industry sales, I'm told, went up 30% and have stayed up. But that was 2006, very interesting time. And what's not well known in history is that we didn't know if this was going to work. I mean, it was a real hell-marry pass. Thanks for having me. Going back to that experiment, those were not wild-type mice. Those were overfed mice, correct? They were metabolically ill. We had both experiments running in parallel, but we got the results earlier for the fat mice. And that was what? So, you had the control

mice and you had the treatment mice, the control mice, but both mice were fed the same, which was a sort of high-fat, high-sugar, metabolically disruptive diet, presumably the usual. Yeah, the so-called Western diet. Yeah. And in that, so what was the outcome of that experiment? How much longer did the respiratory treated animals live? Approximately. I think it was 20%.

Something like that, 25%. And that was initiated at what age? A year. So this is a pretty mature mice. They're middle- age mice almost right there What 30 40 years old equivalent exactly? What dose do you remember? I'm gonna guess something it was a 200 milligrams per kilogram per day per day, but we also had a group on Feeding every other day as well, and those those are not to be pretty interesting as well

That's a pretty high dose relative to did the ITPs that came out that following use dose is that high? So we've done it now in two different doses. We've done 210 times less. Okay. And it actually it works in both. And what we find actually is that the higher dose is acting on other pathways, including the Ampikinesmiotformin-related pathway. Activating it, so it's Ampicon ace metformin related pathway. Activating it, so it's synergistic with metformin? We don't know that, but part of the debate about resveratrol is that if you give

cells in culture or mice too much resveratrol, then it kicks in other pathways. And in this case, it was activating Ampicon ace as well. We published that enough was paid for in 2016, it's activating ampykinase as well. We published that in our first paper in 2016, the supplemental that most people haven't read. But we knew it was activating ampykinase at that high dose. And that what that means is that when you're activating multiple pathways, it's very hard to dissect that. So these days, both in cell culture and in mice,

we're very careful to use the minimum dose to get an effect, not overdose, because it just makes things much more complicated. So that you mentioned that you had a parallel experiment that was done in presumably mice that were not overfed. What did that experiment show? So the fat mice, they were healthier.

That goes without saying, I think, most people know that. In the thin mice, they also were healthier. They had better cardiovascular system, less cancer, whole variety of things that were exciting. It didn't reach statistical significance when they ate resveratrol every day. But if we gave it to them in their diet every other day,

we saw a lifespan extension that tells me a couple of things. One is that it could be that thin fit every other day, kicks in some other pathways that are helpful. We don't know that for sure. We'll be kicking in pathways that aren't helpful, wouldn't it? If I understood you correctly, didn't you say that the mice, the non, so the lean, we're going back to the lean experiment,

if you fed the treatment animals, respiratory all every day, they did not have a statistically significant survival advantage. If you fed them respiratory all every other day, they did. Is that what you said? Yeah, every, so if you only give them food with respiratory, every second day, what you said. Yeah, so if you only give them food with residual every second day, we had mice that were living far longer than just that just to die alone. Were they being fed non-respiratory food on the alternate days? No, they were. Oh, so they were calorically restricted. Right. Okay, so there's now two very so they could have been the chloric restriction that was extending their life. No, because there was

virtual added on to that as well. So there was another control of just being fed every other day. Right. Right. And that came out in a cell metabolism paper in 2008. So from that experiment, one might conclude that resveratrol enhanced caloric restriction, the response to CR. It did. Yeah. So if you had if you had a control group, a CR group fed every other day, and then a CR plus resver plus respiratory group, you would see a dose response. And we did that. And one of the things that might have been happening is that, now, residual is not a very potent molecule. We've got things that are a thousand times more potent now.

But the other problem with residual is that it's not very soluble, and the blood levels both in mice and in humans doesn't get up very high unless you eat some fat. So that Western diet, we had much better absorption than this Lodo-Slean diet. And so it may have just been getting the drug on board, quote unquote, the drug, or it may just be that resverteral isn't potent enough or that we're wrong about this. But the reason I don't think we're wrong about the Surtuins is when you overexpress the gene in the brain, for example, it will extend lifespan and other serotones do that as well. Just going back to that, that's a very interesting idea about the fat solubility.

Was the experiment ever done where you took mice and you fed them within without the respiratory? Because presumably, it's not the solubility of the fat that they're eating. It's the bile acids that enable the emulsification of the molecule and the, you know, whatever the geogonal absorption. So that would be a quick way to show even without a hard outcome if you were just getting more into the cells, right? If the risvera-trol had greater absorption with and without fatty food that's resulted in bile acids secretion. Yeah, well, we didn't measure bile bile acids but we could definitely see by measuring

blood levels that resveratrol was getting on board when we gave some fat in the food. And that's true in humans. That's why if I take resveratrol, I do it with something that's fatty food. Fatty. So some oil, a yogurt works really well. Got it. Okay, well that's so that's elegant. Okay, so then three studies come out there after two of them were ITPs through NIA. One was not. You are on two of those papers, I believe, and one of them, actually two of them were sort of looking at multiple molecules in testing, correct? And I think one of them, I thought it was odd because it looked at a rapamycin, which made sense, and then cymbastatin, and then respiratory. I never understood the cymbastatin part, even like I wouldn't expectcin which made sense. And then cymbastatin and then Rosfaratrol. I never understood the cymbastatin part.

Even like I wouldn't expect that a short-term treatment with cymbastatin would have much of an outcome. What did those studies find, the ITPs? They just showed what we already published, which is that if you give her as a retroling regular food, it doesn't extend lifespan. And why did they, why couldn't that have been overcome? I mean, that seems like they should have given that you'd already put, because you published yours in 06, these were like 08 and 11 or 1112. Something like these were several years later.

Sure. Well, the, yeah, those scientists didn't consult me at all. They just put in the food and went for it. I see. So in parallel to this now, something really interesting happens, which is all of a sudden, a farmer company takes notice of something, which is risk veritall. Was there an IND that gets filed on that, or was this a pure grass application, meaning an FDA, sort of generally regarded as safe pathway?

How was risk veritall taken into a clinical trial with humans, though? Well, what was going on in the commercial side was screening of more molecules. So there was a whole bunch of synthetic molecules that we can talk about. Resverterol itself was considered a proof of concept molecule largely. If it ended up in the clinic, it would have, I mean, as a drug, it would have been great. But what we tried to do was... When your intuition was, it might be, it might not be. Well, we knew it was a terrible molecule.

Because of the solubility alone. Yeah, right. But we... Was it millimolar? Like, what kind of concentrations were needed once it was in the plasma to reach it? Is it a micro-molar or millimolar drug? Yeah, it's a micro-molar, which is, it really, that's not a drug you don't want to go ahead with.

But it allowed us to get into humans and see what would happen. Do we see the same kinds of things? Or could we deliver it on the skin or on the eye? So that was a goal. And there was some very good formulation chemists who were trying to and succeeded in working out ways to make residual absorbable in the body and there were some clinical trials that were initiated under an IND and there was one that was looking at blood sugar levels and that one I think is published now

And what kind of doses were they using in the phase one? Do you remember? that was looking at blood sugar levels, and that one, I think, is published now. And what kind of doses were they using in the phase one? Do you remember? It was massive levels because the scientists that were running the studies, they were worried that there wasn't enough by availability, so they were giving, I think, 10 grams a day, which is-

Was there any toxicity at 10 grams? Unclear, unclear. There was some, in regular people, there was nothing. There was a couple of patients in a cancer study that had renal failure, which happens anyway in these late-stage patients. So that was enough to say, this is not worth it, because this isn't blockbuster drug anyway.

It's a probe at least according to the company that was row running the trials. So they exercised an abundance of caution and stopped that trial in the cancer patients. That's odd. Why was it being done in cancer patients? Well a part of it was that trying to remember, but the person who was running the trials had a lot of experience in it. But I recall that there was some animal data or at least some initerate data that Rizvera Troll could help there.

I see. So I didn't actually realize that at all. I didn't, so you're saying it was in a phase one that they had this questionable outcome and thought, well, maybe it's not worth pursuing this. And we'll move on to the second and third generation variants of these. Well, right.

So the company was bought by a large pharmaceutical company and a large pharmaceutical company, and a large pharmaceutical company looks at Resveratrol and they say, this is not a drug, this is not something that we want to continue with. And so that was certainly part of the decision I'm imagining, but what was, they really were excited about whether the new chemical entities, there were hundreds of them made, there were very potent drug-like molecules that they wanted to take further and they did take those into humans as well.

And I should mention that those molecules have been in mice and rough out a cabo deserves a lot of credit for doing that. And so he's taken the same molecule that went into humans, put them into mice, these same one-year-old mice, and found that those do live longer, even on a regular diet. So, that's why I think maybe it's just where his veritable wasn't as potent as we needed it, and these other molecules succeeded. Now, you mentioned, and I want to come back to this at the end, because it's interesting

you're one of the few people in this space who speaks openly about, you know, look, in the presence of incomplete information, I still take compound X, Y, you alluded to a monorail, you still take risk veritrol. Without having any certainty, is it your hypothesis that you might not be taking enough, but it's unlikely that there's any harm, and you said you take how much about a gram? Well, so my calculation is this.

First of all, I like experimenting. That's, you know, I'm not afraid of death, that's for sure, but I want to be the first person to know if there's a problem, so I'm also keenly aware that what's to happen to all of us if we don't do anything is not pretty either. And so what have I got to lose? It's cheap. I've got buckets of it in my in my basement. There's been never any sign of toxicity. It's been in humans for a long time now. So that is the bigger risk it would seem is what if you're not taking enough? Sure, but you know, one gram, I think if it's not if it's not working at a gram, then it's not working.

I don't think it's worth going higher. But you know, what's important is that we've made advances since then. We're talking about discoveries from a decade ago. We've got much better things now. So I still take my reserteral because I've seen enough data in humans as well that it can protect the heart You know, is it gonna make me live 10 years longer? Probably not not even five, but will it potentially delay cardiovascular disease? Absolutely, so why not? You mentioned in a talk that you I think you mentioned this in the talk, but you take metformin as well. Yeah quite recently probably at least half my colleagues that I talked to.

Yeah, now you're only taking a pretty my colleagues that I talked to. Yeah. Now you're only taking a pretty low dose if I recall from at least what you said in that talk. Is that because of the additive effects potentially of respiratory also activating AMPK? Yeah, you're exactly right. If you're starting to take combinations of molecules like I do, you want to ramp it up. We're going into unknown territory. So I'm in.

Especially if there's overlap in these pathways, right? As you've alluded to. Absolutely is overlap. Anyone who says there isn't is myopic or lying. These are pathways that are additive. So it could be that taking two grams of metformin plus, I'm taking two other molecules would be overdosing it. And I don't want to do that.

So what I do is I start on a reasonable low dose based on all the data that I've read. And then I make sure that I'm okay. I feel okay. My blood tests are okay. And then, you know, I talk to the experts, they're all my friends. And I say, what do you think about going up to one gram or one half gram? So actually, recently, I have gone up to one and a half grams given advice from people who shall remain nameless. Yes, we'll keep everybody nameless. So let's fast forward to some of these other molecules

because the story is just so interesting and feel free to go back and patch pieces of history if it makes things to where we are today. But today we are in a place where there's a company, there are two companies out there that are selling NR, precursor to NAD. They're packaging it with, is it the,

terrestrial being? Tell me what that molecule does specifically and why it would make sense to include that with NR. Yeah, so terrestrial being is essentially just resvertral with some methyl groups on it. It's a more novel, sexier version of resvertral, but that's the, is it any more active in your opinion? So Lenny has told me that it is, but I don't know enough to be able to give a sensible answer.

Okay. So you now have a cert activator with an NAD precursor that are both basically over the counter, right? That's right. So tell me about the excitement in that space. I don't want to get too much into the politics of the companies because they're at the time of this recording. They are in World War 3. So let's just talk about the science and not all the other stuff. Yeah, well, if you take the 40,000 foot view of this. So Lenny is in his 60s now and I'm in my 40s late 40s. And you got to do a calculation. How many years do you have left to not only see what happens, but potentially even be

vindicated. And I'm sure that's part of what I'm thinking. What would let me be vindicated for? For finding that the sort of two ends are important for health longevity in humans. I see. Meaning that following this discovery, some of the air left the balloon, and now there's a resurgence of, hey, no, it really did matter. Is that what you mean by being vindicated?

Well, I think there's a race to see whose pathway is more important. Got it. And the Srituan's and Em Tor and Amy Kainase and some others are in this race. I think it's a silly argument because they're all important. They talk to each other. But, you know, ego is involved in people want to legacy and Lenny's legacy,

as I understand it, can't speak for him directly, but is that he wants to have made a major contribution to human health. And this is a way of showing that within a timeframe that's reasonable, who wants to wait another decade, if I was him, I wouldn't want to. And so I think that that was a time frame that's reasonable, who wants to wait another decade, if I was in my wouldn't want to. And so I think that that was a good way of being able to quickly test the hypothesis in

an area that going through the supplement route. On the other hand, I think because I'd had good experience with pharmaceuticals and I also have more time to wait, I took that route. And so the two of us are heading in parallel. You know, we're very good friends. We talk all the time about this, but it's an interesting, from an historical perspective, that we've got these two guys who are taking very different routes to what we hope

has achieved the same thing, which is to help people in the end. And Elysium, obviously, they've talked very publicly about their goal is to sort of provide a supplement but at a much higher quality, obviously in the United States. I can't speak to this in the other countries, but the regulatory environment here is quite unique in that basically these supplements are quite unregulated. So you're sort of at the mercy of the person who's making the supplement. And obviously having a number of Nobel laureates involved with a company like Elysium in a commitment, a public commitment to use the highest quality stuff, at least a person can

buy this and say, look, I'm not getting crushed bird feathers, which I'm pretty much guaranteed to be getting with half the stuff I buy online. Look, I bring this up because I think half my patients are either taking Elysium's basis or I can't remember the name of the other company, Chroma Dexas, the other thing. Have my patients are probably taking NR. They always ask me, what do you think? Do I say I have no earthly clue?

I'm pretty sure it's safe. So it passes the first test, which I don't think it's hurting you. One of the issues I've always struggled with is, if it worked half as well as it works in the mice, it shouldn't be subtle. You showed in one of your talks an exercise, a contrast between two mice running on a treadmill, and that was actually one of the milder successes. That was like, you know, a 50% improvement in exercise tolerance.

I've seen other studies that talk about an 80% improvement in exercise tolerance. If people were experiencing a 20% improvement in exercise tolerance, I believe we know about it. Do you feel that that's happening and we're missing it? Or do you feel it's just not going to be possible to elucidate that without a clinical trial, even though it's a supplement? I get a lot of emails every day. You might be the single most emailed person on this topic I'm guessing, right? Yeah. Wait till your book comes out. Probably a thousand of them person on this topic, I'm guessing, right? Yeah. Wait until your book comes out. Probably a thousand of them this week, actually.

And I'm not exaggerating. And to any of you who've written to me, I'm sorry, I just can't answer them all. I will try. One of the main questions is, Well, hopefully by having this discussion, let's go as deep as you want into it so that you can say, go listen to that discussion and you'll get

all your nuanced points across. Yeah, I'll go by my book. that's coming out next year in 2019. Do you have a title for it yet, by the way? It's a tentative title. Okay. So the time of this recording, people listening, we don't yet know the title, but David's book will probably be out in the late summer, early fall of 2019. That's right.

Okay. Yeah, so some insights into this NAD world as it's developed. It's extremely hot, but they're selling tons of the material, which is great. There's fit-help people. Now, I get a lot of emails from people who claim and they show me data that they used to do races when they were 40 and then they, now they're 60, they can't win races, they can't cycle cycle but they've gone on this all that in our product or even anime and now they're winning races again. So these are

stories I hear constantly every few days but I can't judge those because these people I don't know there's no placebo but I think that some of this if it if it were true that's what we would expect if that these molecules could help people you would hear these anecdotes, which are here. But I can't declare that I know anything more than you do, Peter, about this, because they're all anecdotes. So that's why we're doing a human clinical trial. Now you mentioned NMN a moment ago. Can you tell us listening to this, the difference between NR, nicotinic riboside, which is

a one-step precursor to NAD and NMN. Well, it's very simple. So, NR is converted by the body into NMN, and NMN is immediately converted to NAD. So, those are the steps. So I misspoke. I actually thought it was NR that was one step away from NAD, and NMN went to NR, but it's the other way around. Correct.

Okay. Both of these can be, we know definitively both of these can be brought into cells to be used as cellular building blocks for NAD. Is there any dispute about the ability to get from high plasma levels into the cell? Yeah. There's a debate. It's not always shattering. It's very academic, but you know, we're debating in the field about which molecule gets

transported and which one doesn't. academic, but you know, we're debating in the field about which molecule gets transported and which one doesn't. And in our definitely gets transported, the question is does NMN get in transported? And I believe in you've written, I think I've read in one of your review articles, perhaps, but I could have been somebody else's that NMN might be slightly more stable than NR. Well, we find that in a lab that if we put them in solution on the bench or leave them on the shelf that NMN is more stable than NR, this is why I keep my molecules and labs molecules in the freezer just to give them some extra shelf life.

And did you imply by what you said a moment ago that there is no dispute that NR at least gets, if you have a high plasma level of NR, it is brought into the cell. Is that diffusion mediated? Is that transported? that NR at least gets, if you have a high plasma level of NR, it is brought into the cell. Is that diffusion mediated? Is that transported? There's a transporter for NR. There's evidence that there's an NR transporter. But either way, they both raise NAD levels quite effectively in the body, in humans as well.

And so... And I'm sorry, I know I asked this already. But when you say they raise NAD levels, you mean mitochondrial NAD levels or cytoplasmic NAD levels? you mean mitochondrial NAD levels or cytoplasmic NAD levels? Oh, gee. So most of these studies have been done on whole cells from blood or tissue. So they're just like PBMCs or something like that? Exactly, exactly. The problem with that is that it's a very dynamic system. This isn't a regular drug that just hangs around and you can measure it. This is actively utilized by the body. So even

if it disappears into some pathway, it's still probably being recycled. So we have to be very careful not to jump to conclusions. If you don't see it in the blood, maybe it was taken up by the muscle or the brain. So these are the studies that are ongoing now. And if you see it in the PBMC, it doesn't mean it made it into the hepatocyte.

Right, this is all just, in an animal, you can do that. We're starting to do those experiments in humans right now using NMR to be able to measure NAD levels living tissue. And when you say NMR, do you mean like MRS or like? Yes, I do. So phosphorus, MRS. And you can measure ATP and AD. Right, right.

So those are really powerful. And you can actually now do it with seven Tesla magnets with people in the machine while they're exercising. So that's where we're headed with this in our next clinical trial, hopefully. In your intuition, David, if you could cherry pick the hierarchy with where you'd want to see the upregulation of NAD most, which cells would you preferentially

be directing it into? Well, it depends on the, if we want a treated disease, obviously we've got to target that particular tissues. So if we're treating a mitochondrial disorder or the muscle, obviously the muscle, for longevity, I would want it to definitely get into all tissues if possible. I think it would help if it could get into the hypothalamus, where there's some central regulation. But what the field has discovered is that all these tissues, not all, but many of these major tissues, are secreting proteins that can induce longevity. So I wouldn't want to

prioritize unless we're talking about a particular disease, like liver disease or muscle wasting. One of the things David, Sabatini and I talk a lot about, I'm actually going to see David for dinner tonight, and I'm sure this will come up as if you could wave a magic wand and make wrap a mison, for example, tissue specific, let alone complex specific, but you get into this. And the same thing with metformin, it probably does have some tissue specificity, probably working more in the liver than it is working in other cells. It seems unlikely that a molecule would have uniform and ubiquitous take up of all cells,

right? Of course, yeah. So with NAD, we don't know. So we're still at the, we're basically still in the infancy of knowing where the NR or NMN would be preferenceately taken up, but it sounds like the MRS studies would, would help us understand that. So that's going to help. And the reason I qualified that statement was that, you know, we're still getting approval for these studies, but that's the plan.

But also there are labs that I'm aware of. I think it should remain nameless as well, for it to protect their own confidentiality, but they are working on tracer studies to be able to give in our NMN and see where it goes in the body and be able to measure each tissue. That will know probably within the next six months to a year. So, you mentioned that what Elysium or Chromodex

are doing is using, they're just basically in a different regulatory paradigm, which is they're using molecules that are generally regarded as safe. They're outside of these IND pathways. You're interested in the same sort of targets, but you're going to go down, I don't want to say more rigorous because that, but it is more rigorous. Let's call this spade. You're going down on much more rigorous pathway.

It's more expensive. That's true. Yeah. I mean, literally two logs more expensive, if not three logs more expensive. So I want to be sensitive to any confidentiality because that's the nature of the work that you're doing now. But what can you tell us about the molecules you're working on directly and indirectly in that space? Sure. In the NAD space, there's a couple of companies.

One is I can divulge, it's called a couple of companies, one is I can go bolder, it's called Metrobiotec, which is here in Massachusetts, and they work on, they've been working for five years on making NAD precursor molecules that are better than these two that are available publicly. And there are ways to improve them, better via bioavailability, better stability, better efficacy, and those are moving into the clinic as well. And then there's another company called Jump's Top Fertility, which is both here in Massachusetts and down in Australia. And we're finding really great effects of these molecules on female infertility or low fertility.

And that's something we're going to be publishing shortly. And is there a type of infertility that this seems most amenable to because going back to your evolutionary argument under periods of stress, we see fertility as one of the first things to go for obvious reasons. But infertility comes in so many flavors, right? There could be an inability to release an egg, there can be aneuploidy, even when the eggs are secreted, there can be uterine hostility, there's so many things that can result in infertility. What does this particular thing target?

Well, so what we find in that, so the biochemical pathway, we've figured out, and we've published a little bit on this, is that there's a protein called Bubber I, which is a kinase that regulates spindle quality. And one of the problems with old eggs is that they don't have nice spindles, and they rip their chromosomes apart when they... Hence the aneuploidy. Exactly, so Down syndrome, etc. Abortion, Aborted fetuses.

But what we found is that this Barbara I is regulated by the CERTI2 protein, which requires NAD. And what we think is going on after chemotherapy or during aging is that the levels of NAD and the ovary and in the egg are low and we're, it's getting this aneuploidy. And that explains why when we give NAD to, when AD precursors to eggs in vivo, the eggs come out healthier, more numerous and are much better at allowing for fertilization and healthy offspring. I don't know much about fertility as this next question will illustrate, but presumably you can still see aneuploidy on the male side. So is there a male fertility opportunity here as well?

I realize it's not as probably as common a problem, but presumably you still have to have a perfect split of the chromosome in the sperm. Yeah, possibly. It's an area that we're looking into. I don't have anything solid enough to be able to say if it works or not, but it in theory yeah.

And you just going back to something earlier, is NMN available over the counter? It is. More recently, I can see it on the internet. Okay. But I, one of the things that I want to bring up is you can find my name all over these products, not basis, they're reputable, but there are others that use my name all over place. Meaning without your permission.

Correct. And Harvard's permission. And so I've sent more cease and justice letters than you can imagine. And they keep popping up. So that's an important point. So if a listener after this talk is saying, hey, David Sinclair sounds like a smart, reputable guy. If his name's on something, I should take it.

What can your name be associated with right now that is, you know, with your permission? Nothing. Okay. So basically, anything that's out there that's over the counter that has your name be associated with right now that is with your permission? Nothing. So basically anything that's out there that's over the counter that has your name on it is not endorsed by you. Correct. Thank you for clarifying that. That would be frustrating.

Yeah, it is, but I try to stay above the fray. It's nice when you can stand on Harvard's shoulders and let their lawyers send those cease and desist letters. It would be nice if that were true when I spend a portion of my salary every year on this. Really? That's a shame. I'm sorry to hear that.

So, fertility is a super interesting angle. I would have never thought about that. That strikes me as highly testable as well, which, you know, one of the challenges of longevity research, as you know better than I do, is if you want to make claims in humans, you better buck a lot because it's going to be, it's almost untestable. So when you look at what other companies are doing like Restor Bio going down the M-Tor Pathway, they're not going after longevity, right? They're going after very, very specific indications that are testable in shorter periods of time in a respiratory failure, things like that.

What are the other disease states that you think would be most exciting if you don't already have plans to go down that from your pharmacologic standpoint? Well, we've been targeting diseases that are rare and have a high on metadneed. So I can't divulge all of those because that's company stuff, but the fertility part,

that's very clear. I'm able to tell you that those trials will begin our next year in IVF clinics, and that will be the first time a doctor looks at those eggs from a woman, you'll probably know that it's working or not. So that's very clear. And when you say company, so we're sitting right now in a headquarters that is the name of, it's what's it called called? Is it life bioscience? Yes. And that's a parent company or sort of, tell me how that fits into a number of

the scientific entities that you're involved in. So life bioscience is a family of companies that uses shared resources and knowledge. There are eight of us right now that work on different aspects of aging, not just an AD biology, but the usual suspects all the major hallmarks of aging that we could list off. Each one of these companies has world leaders and drug development programs in that, so that we think we can quote unquote conquer aging from different aspects, but together we're stronger as one unified company,

life-pile sciences. Earlier you spoke about sort of eight or nine central tenants of aging. We've covered some of them, but I'm guessing that your book is going to go into this in greater detail. Can you rehash what you, or at least as many of those, you're going to recall in the spot, not to put you on the spot. That's a long list. Yeah, sure. There's epigenetic change.

It sells us cell communication and inflammation There's a lot of account this analytics so senescent cells build up There's protein misfolding there's telomere loss and genomic instability. There's metabolic changes So impy kinase and then metformin would address that and then there's responses to what are called amino acids and other nutrient inputs. And those collectively go awry during aging. But what causes all of those to happen? That's something that we've been working on for quite a while.

And you think those are more coupled than they are uncoupled, those pathways, or do you think that, I mean, there are clearly situations in which external stressors can perturb more than one of those. But like, senescence seems somewhat uncoupled from nutrient sensing, doesn't it? It may, but I... And I'm not asking that rhetorically, like, I just don't know. Well, the answer is we think that we've found an explanation for all of these things to happen. A unifying theory? Right. So I've kept it close to my vest for a number of years, but it actually goes all the way

back to the sort of two-in-store in yeast. And hopefully the listeners who've stuck with this podcast are still with us because they will punch you. Yeah, they promise you they are with us. So the punchline is that, so this is all off top of my head here. We haven't published this yet, but I'm going to tell you my thoughts and your listeners. So the genome is digital information. It's very easy to preserve. The reason we went from analog to digital in 2000s, DNA is four letters, it's digital, it's easy

to replicate, it's easy to store, you can boil it, it's very robust. And so what we've actually come to discover is that the genome is fairly intact in old people and old animals. We've sequenced the genomes of lots of old mice, and all the genes are still largely intact. So what's going wrong? Well, the other part of information that you inherit from your parents is the epigenetic information. Okay, and I use term loose sleep, but basically it means what's the pattern of gene expression, which genes are turned on and off at which time. And that is analog information. That has to be analog because, instead of just being a single code, it has to operate in three dimensions, actually four if you count time. And so that's an analog system, and it's constantly adapting to what we eat, what we drink, if

we run, when we sleep. And you have to turn jeans on and off all the time. But that pattern of gene expression that's set down when we're young, because it's analog, analog information doesn't last very long. Anyone who's had a record player or magnetic tape knows that these things don't last. And that's the problem, think with aging is that we don't lose the digital information. So the compact disk of our lives is still intact when we're old. But it's as if we've got a scratched CD and the cells don't read the right genes at the right time anymore and they lose their identity. In fact, if there's an analogy, which is called Warrington's landscape, where in the

1950s Warrington drew a picture, it's a beautiful picture of some hills, it's a mountain scape, and cells actually rolled down the mountain scape and landed different valleys down below. And that's two, before we had, he had access to the genome. That was his way of saying, this is how cells know what they are. They land in these valleys and they stay there. But what I think is happening during aging is due to the vibration of noise over time, we lose that pattern of gene expression, we lose that inflammation, epigenetic inflammation, and those cells or those marbles in Wattington's

landscape, they jump over into different valleys and lose their identity. So your neurons are not functional, like neurons anymore, you'll live your cells in more like neurons. And we see that in our lab. We're just writing up a couple of papers right now for this. And we're able to actually manipulate the epigenome in cells and in mice and have a look what happens to those animals. And the prediction is that you get all the homoxid aging. You know, the challenge with this entire space is

you think back to the time in the 1950s when he made, when he created that analogy. And it's in some ways, it's amazing that it could still be relevant 75, 80 years later, whatever it is. On the other hand, it, it humbles you to realize how much more has been learned about that process in that time. And sometimes I think about it because you and I are interested in the same problem that I'm worried I just don't know anything.

You know, I'm worried that in 10 years, I'll look back at my hypotheses and my, or not even my hypothesis, just my understanding of the current state of the art today and think, you know what, that was directionally right, but it was so oversimplified. And oh my goodness, like, you know, so it's sort of

like we're back in this problem of time, like we're going to run out of time. And I mean, how confident are you that, because you and I are almost the same age, like how confident are you that in our lifetime, we will see step function changes in human longevity. And to put this in context, there really hasn't been a step function change in human longevity. And to put this in context, there really hasn't been a step function change in human longevity probably since the introduction of sanitation. I mean, everything has been quite incremental. Maybe antibiotics, vaccinations, antibiotics have probably been the last step function change. Will we see one in our lifetime? How confident are you?

I'm getting more and more confident. Honestly, when I started in this field, I thought we'd probably not see the type of technologies that I'm seeing now. It confident. Honestly, when I started in this field, I thought we'd probably not see the type of technologies that I'm seeing now. It's making my head spin, not just in the technologies, but also the investment and the number of people working on this now. This was the back order of biology when we started. And there's been some new results,

which I'll just hint upon because we haven't published, and it's very early, but I've seen, it sounds like a scene out of Blade Runner, but I've seen things you wouldn't believe, and it's maybe not that dramatic, but let me go back to the compact disc analogy, you've got the scratched CD, how do you find the polish, what is that?

Let's go back to the yeast analogy, what causes those scratches? Why do you get loss of gene regulation? Anyone who is paying attention early running this conversation will remember that these DNA breaks in the chromosome, broken chromosomes distract the cirro complex and they move away and you get the expression of genes that have no right being on. Because the circoins have lost, they're distracted from the deactivation function and they're dealing with the repair function. Exactly.

So using that, what we've got a lot of evidence for now is that something very similar, if not essentially identical, in principle, happens in mammals as we age. What that means is that insults to the genome, and one of the major insults is a double-strand break, but they're probably others, cause these proteins, certunes and other factors, I'm not saying only certunes, but factors that control gene expression, silencing and other things. Have a dual role, we know, in DNA repair and other things, such as responding to stresses, heat, whatever. But this is the cell's way of coordinating gene expression changes, hunkering down during

times of adversity and going off to repair the system, which in this case, we study DNA breaks. And that's a beautiful system when you're young. It works great. You get exposed to cosmic rays or you go out in the sun, you got lots of DNA breaks, eventually, these proteins will go, repair those breaks, and then

go back to where they came from to settle down the response, to turn off the inflammation, to turn off the inflammation, to turn off the DNA repair when it's not needed. But the problem we think is it's antagonistic pleotropy. Okay, so Peter Medeware and the other brilliant scientists in the 50s speculated I think correctly is that things are really good for you when you're young, come back to bite you and the ass when you're older. And I think that's what's happening here is that this response to these stresses like a, end up not just distracting these proteins, but end up disrupting the actual structure of our chromatin. And these proteins don't

always go back to where they came from 100%. Do that for 70 or 80 years. And it's not surprising that the genes that were once perfectly programmed and turned on at the right time lose their ability to do that. And we've got remnants of that program when we're a 70 and 80. But what's exciting is that information is still there to be accessed. The question is, how do you get the cells to remember to access at the right time?

What's that polish? And I think we're pretty close to finding that. If you had unlimited resources, and not just financial resources, but sort of metaphysical resources like any experiment would be ethical, you could do something that today no IRB would approve. Is there an experiment that you would wish to see done that could accelerate our knowledge in this space log fold? You mean staying within ethical? Yeah, but for example, to do a human experiment in longevity

would be ethical but impractical because of the duration of time. If I gave you a time machine and an infinite amount of resources, tell me with the most elegant experiment you can think of that would just leapfrog our knowledge. The experiment that needs to be done, whether it's with metformin or other drugs in development, including these rapologs and the NMN and NAD precursors, it should take a group of 5,000 people. That would be sufficient and just give them the medicine and wait three,

four years. And you'd know from that number of people that you're changing the hazard ratio, the mortality rate You'd have to start with people probably in their 70s. I think that was the calculation that I did But you don't need to wait a whole lifetime to know that these things were so this is basically near-es argument Yeah, exactly but near is not doing mortality as much as he's doing health span But it's exactly right you do enough people. I mean, it's gonna cost tens of millions of dollars But think of the trillions of dollars that would be saved if we can prove this. But if I'm going to play devil's advocate for a moment, what if by doing that experiment,

we're missing the opportunity, the window of opportunity for these drugs to act. In other words, you know, we know that caloric restriction is less and less effective the longer you wait in the organism, at least within mice. And that's what made RAPA mice, and so interesting, is you could, it was, it blew everybody away, that you could start this drug on mice that were 600 days old, and they still had, you know, 9%, 14% increase in lifespan. If you had more time, do you think we could get a more clear answer starting earlier, acknowledging that you'd have to wait longer if you wanted to use a hard outcome.

Do you worry that we would risk doing this in people in their eighth decade that it might not work, but that just tells us that it doesn't work late, not that it doesn't work period? I don't worry about that. We could, with enough capital, we can money invested, we could do a multiple different experiments.

We could do people in their 50s, 60s, 70s and 80s. So, you know, dream with me. The other thing that makes me optimistic is it's not just revenues that works late in life. We've got results in my lab now that we'll be publishing that if we start even later than 600 days in a mouse, which is what closer to us. 75 days. We can still extend lifespan. Using what I know you want. This is not, I can tell you we're using the NMN.

Okay. So there are multiple ways to add like... When you use NMN in the lab, are you also using like a PT analog or a respiratory analog, or are you able to just use NMN and see? Just the the natural molecule will work when they're gearing up to do the. And this is not in high fat over fed animals. This is in wild type animals. We just put in the drinking water. Oh, because NMN is more soluble.

Exactly. Interesting. Yeah, but we've got better molecules that we're now testing. We think that we can beat or again try to beat rather mice and maybe a combination together. Do you think there's overlap in those pathways? Yeah, yeah, you were thinking the same thing. Yeah, I'm going to do the combination. Yeah, yeah, yeah, and I want to take the combination together. Do you think there's overlap in those pathways? Yeah, yeah, we're by thinking the same thing. Like, yeah, I'm about to do the combination. Yeah, yeah, yeah, and I want to take the combination one day.

Well, we know actually have some early data. I don't want to scoop myself, but we're able to now genetically modify adult mice with AV, the virus, and then associated virus. And we can now genetically change a mouse. So we've just put in all seven sort of genes into an entomise, in old mice. You've put the seven human certain genes into mice. Oh, seven mouse genes into mouse.

Okay. But that sort of experiment would have taken a decade to do just a few years ago, but now that we can deliver genes, we can do very good experiments. Not only that, we can also do that multiplexed, we can do combinations of genes and combination of molecules. So we even experiment. Yeah, you're stacking the matrix now. Well, yeah, I mean, we should be doing this. It's just a matter of resources, but I think we're now at a point in the aging community where these combinations need to be tested. It's the question that's on everybody's mind, what happens if you put them in combination?

Are they better or worse together? And so what we've done is we've put all seven sort of genes into a mouse and fed them some NMN to give them the fuel and the genetic requirement. And interestingly, there are additive effects when you do both of those things. And this is how we're able to see these dramatic effects in these old mice. So is your optimism towards the NAD precursor space and these are two in activating space equivalent? Or do you and you see them as necessarily parallel pads? Um, they have different uses.

The NAD is where I'm mostly focused on now. And is that because of the observed age related decline in NAD? In part, but it's also because in theory, all of the seven sort of twins should be good, and their lack of NAD could be the main problem that's going on in older people. And so the idea is that instead of just activating one sort of two in, which is what Resvertral did, we think, you can potentially activate all seven of them and replenish what's been lost over time. And I think seven is probably better than one. You mentioned at the outset of this discussion

that you were four years old when you became aware of mortality. Was there something that occurred that made you aware of mortality at such a young age? I had an unusual grandmother who was really honest with me. She would never lie and any question I asked, and I was quite... And you were probably very precocious and curious.

I suppose I was, but yeah, she would never lie. So my question that most kids have is, are you always gonna be around? She didn't even, couldn't call a grandmarsh, wanted to use her first name, which is Vera. So Vera, you're always getting around. Of course not, I'm gonna die. Yeah, but what about mom and dad?

Yeah, they're gonna die. What about my cat? is going to die. And then of course you're thinking, if they're all going to die, what's going to happen to me? And then, but it turns out all kids go through this, has been studied extensively that between the ages of four and seven kids understand that there's death. And at first they're in denial, they're saying, well, maybe those adults will die, but my teachers and my parents, they're not going to die. But then by age seven, it's undeniable. All kids know that everything around them that's living at one day will die, including themselves. What's very interesting when that happens at age seven is it's buried deep in the subconscious

and you very, really think about it until you have to. Because I don't think as a species we could get by for all running around, oh my god, I'm going to die one day. So seven-year-olds onward till about 50, you try not to think about it. That's pretty common. You start looking at yourself in the mirror at 50, you notice your teeth are wearing out, you just kind of feel a little bit of age, you see some wrinkles, you think, oh yeah, crap. This is really happening.

And you know, you think more and more about it. I'm unusual because I work on this every day so I'm always thinking about it But I think on average when I talk to people especially in their 20s and 30s It's not something that is on their radar because it's gonna be something happens in the distant future that they're not even looking at Do you have kids? I have three. How old are they? They are 15 13 and 11. How do you explain what you do to them? I was just as brutal to them as a microsite. I was too.

Yeah, and actually I saw my oldest daughter go through this, actually all my kids, but it was more dramatic to see it for the first time myself. And I gave her Ted talk about this. When I told her that I was gonna die, she burst out crying and for a week couldn't sleep. It was traumatic. And you know, maybe I'm a cruel parent, but I also try not to be as my kids either. But what I

saw with her was what even I think most kids do, including myself, was you just can't think about it. It drives you nuts. You won't sleep. So she forgot about it and we've never talked about it ever again. And if I talk about death, she says, shut up. Don't want to hear about it. And she probably won't think about it in a big way until I'm old or her grandparents are dying. David, this has been a really interesting discussion. I wanna be sensitive to your time.

I think the listeners of this podcast will be upset to know that we're only going for about, you know, an hour and 40 minutes instead of the usual three or four hours. But where can people find you on social media? And how do you like to interact with people that have questions besides adding to the list of the thousands of emails you get a day? Yeah, emails, I can't reply to everyone, but on social media, I'm now on social media, I find that's a good way to communicate with people. And so I have a Twitter handle, which is David A. Sinclair, an animal so on LinkedIn, but we're putting together a social media page so we can have a discussion.

It's now there's so many people interested in this. Any information that's locked in my head seems to be on demand. So that's how I want to reach people. This book is coming out which has me regurgitating and vomiting on the page, everything from what we've learnt in my life with my kids all the way through to understanding why we age in this universal hypothesis that we're putting forward. And then the consequences of what happens when we do this.

Now it's not a question of if anymore. It's a when it's going to happen. What happens to planet earth, what happens to humanity, what happens to your family as this starts to come out. And some good, some bad, what do you have to get ready for economically, socially, and it's all going to be in there. Well, I really look forward to reading it. You know, I'm in the midst of writing a book as well.

And I'm guessing you struggled with the same thing I'm struggling with, which is not just the writing of it, but more importantly, you almost don't want to hand the thing in, because you know that the day you hand it in, there's something new that you're going to want, you're going to know something more. And so as we were talking about it before we started the podcast, like you submit these books a full year before they hit the press. And then of course, it basically is a static document until, you know, sure, you could have an online updated to information.

But did you struggle with that? Especially I think, you know, people like us, I think have a little bit of a humility around the half life of facts. And that sounds like a very, you know, sort of pompous thing for us to say, but I just think we have the luxury of knowing that basically whatever we know today

is, you know, quite likely to not be entirely true tomorrow. How did you cope with that? This is now just a very personal question for me as I'm struggling with this phenomenon. Well, yeah, you're a perceptive guy, Peter. So I'm also one of those guys that I'm at the podium about to give a talk to a thousand people and I'm still changing my PowerPoint slides. I'm just obsessed with perfection. And that's one of my downfalls. The same thing with the book. I've been writing it for 10 years and it's needed updating, of course, even to the point that last night I was editing it. I've already turned it into my editor three weeks ago. You're a nightmare to the editor.

And she's received probably seven updates already. And I'm going to stop, I promised, but every time I wake up there's something more really interesting to add that I want to put in there and You know a year from now. I think it's gonna be an even better book than it is now But I'm really happy with everything I've gotten down on the page because when I talk to people people write to me every day I'm able to answer questions that I think are burning in a lot of people's minds. Well, David This has been great. I find it it's sometimes hard to talk to people You don't know until the day of the podcast, which is the case here But I found there's a common language that we speak or maybe

a common passion, not a common language. This is a topic that's new to me, but anyway, this has been really exciting and I can't thank you enough for your time. Well, I appreciate the opportunity and I really enjoyed it too. Thanks. You can find all of this information and more at pterotiamd.com forward slash podcast. There you'll find the show notes, readings, and links related to this episode. You can also find my blog and the NerdSafari at pterotiamd.com.

What's a NerdSafari you ask? Just click on the link at the top of the site to learn more. Maybe the simplest thing to do is to sign up for my subjectively non-lame once a week email where I'll update you on what I've been up to, the most interesting papers I've read, and all things related to longevity, science, performance, sleep, et cetera. On social, you can find me on Twitter, Instagram, and Facebook, all with the ID, Peter, Etienne, MD,

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