Is This The Most Advanced Anti-Aging Analysis & Epigenetic Diagnostic Test Of The Future?

On this episode of the Ben Greenfield Fitness Podcast, I'd actually like to start with acceleration of aging. What are you finding are the things that seem to really influence biological age deleteriously?

You know, your past life experiences are coded in your genes and your genes are optimized for a particular pattern that we look at for genetics.

Exercise, particularly aerobic exercise, has more of an impact. Whereas if you're younger, I think that things like sleep, closure to pollution, exposure to environmental toxins are a little bit better.

Health performance, nutrition, longevity, ancestral living, biohacking, and much more.

My name is Ben Greenfield. Welcome to the show. And today's episode is with a really smart pair of guys who have been studying up on so called epigenetic aging, Dr. Dan Stickler and Ryan Smith. We're going to be talking about this true age epigenetic test. So it's kind of fascinating. Hope you enjoy. So that being said, enjoy. Well, I've talked about ways to measure your, your age on this show before. I've talked about all these different telomere tests and briefly mention things like methylation clocks or Horvath clocks, and really have several podcast episodes out on how we can track anything that we might be doing, say nad or peptides or sauna or sunshine or anything else when it comes to living a longer time. But the whole field of being able to track and measure and optimize biological age is, in my opinion, a field that's kind of rife with confusion fusion and a dizzying variety of testing methods like telomere analysis or methylation clocks or anything else that's popping up. And I've recently been looking into this concept of DNA methylation testing and epigenetics and how we can learn from that really what our biological age is. And I decided to interview for the show today two people who are kind of at the cutting edge of this whole biological age testing sector to get their take on where we're at and to really educate us on how we can track in a scalable, sustainable way, our biological age. So my first guest on this show, you may have heard him before because he's been a couple shows in the past in which we've talked on all things peptides. He's a real expert on peptides, but he also is in the biological aging sector and his name is Ryan Smith. Ryan, what's up, man?

Hey, Ben, thanks for having me.

Yeah, no problem. And my other guest you may have heard of before because he's well known in, I guess maybe you'd call it like the functional medicine industry or the genetics testing industry or the genomics analysis industry. And that's Dr. Daniel Stickler, who's the co founder and chief medical officer at apeiron. And he is a physician to a ton of high performing execs and entrepreneurs. He's also an author, a speaker, a blogger, podcaster. Dan, welcome to the show, man.

Thanks for having me, Ben.

All right, folks. Well, there's a lot of different areas where we could, we could kick this off, but I think just to make sure that we get our terminology straight here and Ryan and Dan, as is the case, it's always a little awkward doing a, you know, three people on the show and you guys deciding virtually who's going to respond to which question. But I'm going to ask it anyways and let you guys decide who's going to dive in. And that is differentiate for people. Buy a chronological age versus chronological age.

I can take that one, Ryan. I actually have a little bit of a qualm against using either one of those because what it does is it compares you against the norm of the population, which I don't necessarily agree with. So chronologic age is nothing other than an arbitrary number, number of years since you've been born and you can take a 70 year old and compare them to a 50 year old and there are norms of that in the population, but everybody's going to be very different in that regard. And we also see this in biologic age, which we're actually looking at the aging of the cells. So it's like a clock of the cells, chronologic clock of the cells, and you can accelerate, you can decelerate. And more recently we've seen that there's the potential to actually reverse that clock. Again, it's not a great metric to use. It's a great dynamic to use. So when I say a person's biologic age is 60 this year and next year it's 58, that's a very positive change in that age. But it's still a relative number that you're comparing to a norm. And I just don't like comparing to norms. It's not as accurate as I would like. I would prefer to see people get a health score and say, hey, your health is 96 versus somebody else who's 52. But unfortunately that's what we're using right now and that's the best way we can do it until we develop something that can truly get a more in depth measure of that.

Okay, what do you guys think about Telomere analysis, because up until that point, I mean, that's what a lot of people have been talking about. And you know, I interviewed Bill Andrews from Sierra Sciences, and he talked about tests such as life length or Renew diagnostics or Spectracel as being decent ways to analyze the rate at which your telomeres are shortening. And I wonder if you guys have a take on telomere testing.

Well, I've done telomere testing. I've used life length for the past 10 years in my clinical practice. But I've gotten more away from it because we don't know for sure whether telomere age is a cause of aging or it's a metric of aging, you know, like gray hair. Does that really indicate that it's causing you to age? We don't know 100% on that. We suspect that it's relevant there and right now, and I know Ryan's in the same court that we are in clinical practice, that we just don't have 100% testing right now. So what we do is we get all the different potentials for determining agencies and we start putting these together in a nice computer algorithm and start seeing what is truly the marker that we should be looking at or the group of markers.

Yeah, and if I might jump in there as well, the telomere question is actually a really, really big one. Just because a majority of the people who have been involved in this space are conceptualizing this thing, the HAPIC limit and how it affects health and disease. But there's some really, really good information about the interplay between telomere leng and outcomes of physiological function or biological age. And the telomere testing and telomere length is one of the. Has numerous large scale epidemiological studies, but there's only been a weak negative association with markers of biological age and life expectancy. And so a lot of people postulate that that's because it might not have a physiological effect until the telomere length is critically shortened. But it definitely has some implications as we talk a little bit more about how this test was created in future areas of interest in terms of stem cell telomere le and other things. I'd be happy to go into that a little bit later.

Yeah, well, I mean, you could dive into it right now while we're talking about telomeres if you want to go for it.

It's a complicated thing because, you know, in order to, in order to get to why it's, it's so exciting, you really have to divide this Calculation of epigenetic age into intrinsic and extrinsic epigenetic age. And, you know, that becomes a very, very difficult conversation because, you know, it really all started in 2013 with two different types of clocks. Dr. Horvath, as you mentioned, has sort of been the pioneer and the leader in this. Dr. Greg Hannam, another biosthetician, came out with a clock at the same time. And over the course of several years, they realized that those had different powers in predicting different things. And ultimately they realized they were measuring sort of two separate things, both of which are incredibly important to biological age, but have different outcome metrics. And it's sort of a complicated discussion, one that we can sort of talk about when we talk about some of the interventions with Dr. Fahey's TRIM trial. But one of the things that they found is that the telomere length of stem cells is maybe one of the most exciting things and might underlie the entire biology of methylation. And so whenever we talk about sort of the process and what methylation is and how it happens, telomere length will come back into focus.

But it's the telomere liked on stem cells. Is there a way to actually test just the telomere liked on stem cells? Because, you know, if I get a telomere test right now I'm doing, you know, typically a finger stick blood drop, which I assume is not just testing the telomere length of stem cells.

Yeah, as far as I know, white.

Blood cells, I mean, that's what they're typically extracting the DNA from. And it's an indirect marker of the stem cells in that regard.

Okay, so they're just kind of basically extrapolating from the telomere length on a white blood cell what the telomere length on all the other cells in your body would be.

Right. And, you know, I like life length because they do the shortest percentage. That's the reason I've always used them. You know, as Bill Andrews said, if you're in a room full of dynamite, do you want to know the average length of the fuses or the shortest?

Right, exactly.

And so that's why we've always stuck with them.

Okay, got it. So how does this concept that seems to be coming up more and more these days of testing epigenetic markers compare in contrast to something like testing telomere?

Like, well, there's a. I mean, there's a whole host of. I mean, this is the holy grail right now. I mean, everybody's looking for the marker that can determine our True biologic age, our, you know, our true time to death and all of this. So, you know, one of our, one of our board members, Brian Kennedy, former CEO of the Buck Institute of Aging at Stanford, he is aggressively looking for it, along with about probably 40 other labs around the world that are trying to find which markers are relevant. And right now we're in our clinical practice, we're in a private clinical practice, and we're doing things like epigenetic age, we're doing glycan age, we're doing brain age by qeeg, we're doing blood markers, we're doing telomeres. So we're gathering as much data and that's one of the things we're working with Ryan and their company on, is we gather the data. I mean, we're Even looking at VO2 max changes over time and putting this all into that database so that we can have that AI algorithm that can start looking at this and seeing which are the relevant markers and how they interact with each other.

Definitely. And to add on to that as well, taking a step back, the reason that biological age is so important is because it provides one single objective measure that is the largest risk factor for a lot of age related diseases. Dementia, osteoarthritis, you know, metabolic disease, cardiovascular disease, all these are age related diseases. And so if you can define a biological marker which is really indicative of functioning, you can test interventions against it. And that's powerful because then you can extend health span, you can extend lifespan. And using that as sort of an objective measurement technique allows you to develop the tools to really make a difference in health span.

Okay, so before this podcast interview, you guys sent me over a test. I think it was called true. What is called true age or true diagnostics?

Yeah, true age is the test. Yeah. But by true diagnostics, okay, I did that test.

I didn't get the results back yet. But how does that differ from telomere testing?

So I think that the biggest thing, the difference between telomere testing and DNA methylation testing, one of the biggest differences, how they're tied to outcome measurements and then also how predictive they are of actual age. Telomere testing, like I said, doesn't really have a huge statistic statistical significance to markers of biological age or even life expectancy. The methylation age or methylation age calculated by these methylation DNA markers is sort of the opposite. It has really, really tight correlations to particularly aging and death and death prediction. And so it's a little bit, I would say, more functional. It Tells a little bit more about risk to certain types of diseases, but also gives you a better sort of overall readout of sort of where you're at in your health. And actually, you know, a lot of the telomere studies have been sort of knocked, you know, in the. In this sort of integrative medicine community because they can change so fluidly, you know, and they're a little bit less reliable. You know, sometimes they will jump up or go down, and really no one has any idea why. But the epigenetic aging calculators are much, much more specific. The amount of data that is used to. To make these tests is absolutely much more expansive. Looking at hundreds of thousands of copies of DNA, and as a result, you can get really, really precise measurements which then predict risk of a lot of different things.

Okay, so I want to jump in here real quick because I'm a little bit confused, because DNA methylation or histone modification are two ways that I know that DNA is influenced. But. But epigenetics is different than that. Like, when you're testing epigenetics, you're not testing changes in the nucleotide sequence, right?

No, in the epigenetics. Epigenetics is the aspect of. And methylation is a big part of that, but there are a lot of other processes that can occur, but it's in changing the way the genes are expressed. So the genes are always going to be there. It's just some of them are turned on, some are turned off, some are turned up, and some are turned down. And there's a difference that I think we need to make here in histone modification versus CpG methylation. So CpG methylation is methylation of nucleotides right. On the DNA sequence itself. And most of these usually occur in promoter regions of genes. So that's where the genes are turned on and off, predominantly. Most lifestyle patterns have. They're hard to shift when it's a cpg methylation pattern versus a histone modification, which is very malleable to lifestyle patterns. So differentiating the two types of methylations there, I think is important.

Okay, and so what is epigenetics testing for? What is epigenetics testing, testing for? I guess I should say.

Well, there are a lot of different types of epigenetic testing, a lot of things that are associated with histone modifications. But the one that is most related to biological age is this methylation testing, where they test whether or not these cpg sites are on your DNA. Sort of in these promoter regions are methylated or not. Usually, methylated is associated with inactivation of a particular gene in comparison to acetylation. And so you're actually testing those methylation spots on your DNA.

Okay, got it. So when you say cpg sites, I assume you're talking about that would be the region of the DNA where you have a cytosine nucleotide followed by a guanine nucleotide, and what you're looking at are changes in that specific region.

Absolutely, yeah. The cpg sites are sort of the cytosine and then a phosphate and the guanine. They're called. They occur in high frequencies in certain genomic regions, and they're really called CPG islands. And those areas are methylated at a very, very predictable rate, which infers that there's some underlying biology which is controlling this throughout aging.

And so with the rate of methylation at those sites, then indicate excessive aging if that rate of methylation was accelerated and decreased rate of aging if that rate of methylation was low or something else.

Yeah, absolutely. And it's not necessarily increased or decreased methylation, because in some areas, increased methylation can mean that you have a higher epigenetic age, and in some other areas, an increased methylation can mean you could have a lower epigenetic age. So it's all about correlation to these differences, where each site is correlated to chronological age. But absolutely, these are the sites that are measured to give us that reading and very, very predictable measure of biological age.

Okay, so someone has actually researched the type of methylation that occurs at the CPG locations and determined that the type of methylation that's occurring is going to correlate directly to your biological age.

Correct, and with extreme accuracy. And I know that Dr. Stickler can compare to a lot of other test, but, you know, the typical margin of error for things like Dr. Horvath's tests are around 2.6 years, which is incredibly, incredibly accurate. And it has a lot of implications in things like forensic testing, where they might find someone's DNA at a crime scene and need to know how old they are. Or even, as we talked about earlier in death prediction, because if someone is 30 but has a biological age of 40, that's probably not a good sign for their overall health. And so other companies, like health insurance companies, for instance, are really interested in. In this metric as well, because it can really help predict death. And then therefore, they can decide how much to charge or whatever that might be. And so it's a very, very predictive measure, especially when you compare it against all the other metrics which are available now, proteomics, metabolomics, or even telomere testing.

Okay, now, like I mentioned, I haven't actually seen my results from this true age test. I think they're coming in next week. But is this like a telomere test where it tells you, whatever your chronological age is 40, but your biological age is 43, or chronological age is 40, but your biological Age is 27, or is it more of a result that says something like this is when you're going to kick the can?

So at the moment, it does not predict death, and there are definitely some ethical implications as it revolves around that. So this is basically a reading of your current cellular biological age. As Dr. Stickler said, it's a measurement of how your cells are, how old they are and how they're behaving. And it also, in addition to that, also talks about some of the factors which are associated with these different changes so that you can make, hopefully, some lifestyle or medicational interventions to try and live a healthier life and decrease that biological age.

Okay. And that's because we know that epigenetics are heritable changes in gene expression. They don't alter your primary DNA sequence, but they are influenced heavily by lifestyle factors like nutrition or smoking or alcohol or physical activity or. Or even things like relationships, meditative practice, et cetera. These are all modifiable methylation patterns that you're testing.

They are modifiable. And like you said, and I like that you said that they're also heritable. And this is important because some people are born with methylation patterns that can indicate that they're older than their baseline. Just from the heritable aspect of it. I mean, most people don't realize that really the most critical timeline in your life are the three months that your parents are in the preconception period, right before conception, during your mother's pregnancy with you, and in the first two years of life are very critical for developing methylation patterns on these CPG islands, especially, they will set the stage for the rest of your life. Now, throughout life, you also gain and lose methylation patterns through different lifestyle factors. But we like to say that some of these methylation patterns are written in pen and some are written in pencil, because the ones that are written in pen are really hard to change and erase. They're kind of fixed patterns that accumulate over time versus the ones that are written in pencil, which can change. And then. And most people used to think that from an epigenetic age standpoint, it was A purely one way linear process. And we've learned just with the Fahey study back in September, it really changed the mind of a lot of people in understanding that we can actually reverse age, the cellular aging process.

Got it. And for people who want to get this straight, I mean, the way to think about it is the word EPI that comes from, from Greek and it means above, translated from Greek. Genetics is just the study of your DNA. And so when we talk about epigenetics, that's a study of things that might go above and beyond just the, just, just your genome. You know, the DNA that you were born with. These are, are things that can be modified that go beyond just, you know, like crispr, you know, going in and trimming would be an example of how to directly modify the DNA. But we're talking about modifying things that as the word epigenetic supplies or refers to, go above and beyond just DNA, Right?

Correct.

I just wanted to add a little bit. You know, this is one of the big parts because, you know, a lot of people are scared to get their DNA sequenced because they might find an APOE 4 variant that predisposes them to Alzheimer's or something like that. And they don't want to work with something they can't change or know something that they can't have any effect over. And the good thing about these methylation changes is that we're sort of finding out that over 60% or around 60% is modifiable. And so you're still controlling the majority of your genome and the expression of these things which are linked to aging processes.

So the tricky part here for me is that from what I understand, there are literally millions of these cpg islands on the human genome. And so when we're talking about testing and calculating your biological age, that seems like a hell of a lot of sites to test. How is this done? Is this done with like AI or some kind of computer algorithm or, or, you know, when you're researching? I think I do so many tests, I forget. I think it was a drop of blood. It was a drop of blood that I sent in.

Yeah.

Okay, so when that drop of blood is received, how are you testing millions of different CPG locations?

Yeah, and I'll talk about this a little bit. So our company, True Diagnostics and the test TrueAge, utilizes a microarray based platform. And so what we're doing is we're isolating DNA, we're doing something called a bisulfate conversion, which allows us to test where these things are methylated and then what we're looking at is over 900,000 different locations on the DNA using these microarray beads. And so what we're doing is we're complexing different parts of the DNA to these beads. And whenever we look at those through sort of spectrops, could be methods, we're able to see if they're methylated or if they're not. And so you're testing many, many copies of the same DNA and you're testing 900,000 spots on them. So the amount of data that you're generating is extensive, but it's looking at a lot of different regions. And usually that's also an important part because the algorithms to read these things that are reporting out these biological ages actually don't use nearly as many data points as you would think. You know, for instance, the first Horvath clock only looked at 353 different locations on the DNA. So what happens is whenever you look at these wide genome studies, you correlate those to variables like chronological age, then you pick the things that are most highly associated with it, which gives us the ability to have these biomarker estimates which are extremely, extremely precise and accurate.

Okay, got it. So when I send that blood in, you're testing a pretty large number of spots on the genome that are directly correlated with, I guess, I would say, expression of certain genes. Like, you know, for example, your eyes are going to express different genes in your hair. And as you age, expression gets harder to regulate. And that's where a lot of chronic diseases set in. So you guys are looking at how good a job your DNA is doing at expressing certain factors.

It's mainly just a correlation between methylation sites and outcome metrics. You know, that that's really what these computer learning systems are doing, is that they're using large amounts of DNA, you know, information to compare to this, this health information, or things like chronological age. And so by looking at such a wide variation, you can really have some really good predictive algorithms and, you know, some of these things. For instance, if you wanted to run Dr. Horvath's original algorithm, you could only look at 353 spots on the DNA. But the test itself is different than the algorithm to look at it. The test sort of establishes where you're at and what sites on your DNA are methylated. The algorithms are really the glasses or lenses to which you read that data and turn it into actually something useful.

Okay, now there's a lot of talk right now about senescence, and especially you Know, something like immunosenescence, when your immune system kind of becomes weaker and less functional as you age. And that's often seen, I know, in a blood analysis, by having a lower number of T cells or a lower number of naive T cells and a higher number of senescent T cells. When you guys are getting this data back in, are you able to look at anything like cellular senescence?

Yeah, absolutely. By looking at so much of the DNA, it's all about creating new algorithms to read that. And if you can correlate, for instance, these methylation sites to senescent markers, then you can come up with new tests for senescence. And that's actually what we're doing. In addition, there's already, you know, IP out there that looks at these same epigenetic methylation markers and correlates it to things like telomere length. And so as we, as we sort of progress down the exploration of this as a diagnostic, you're going to have this, the same measurements, these same methylation measurements, be able to report out a lot of different things. And we are working one on senescent cell burden, where you can just look at the constituent of your DNA and then figure out how much of a senescent burn you had and possibly how much senolytic cell therapy might be able to help.

And that's one of the things we're working with Ryan on, too, is we measure. We use UCLA's lab to do senescent and naive T cell ratios, and we're plugging that into the algorithm with Ryan for helping to get more accuracy on.

That T. This is also a really good point to talk about this difference between intrinsic versus extrinsic epigenetic age. As I talked about those initial two algorithms, Dr. Hannams and Dr. Horvath, whenever they started to validate these algorithms, what they saw is that Dr. Horvath was really, really good at predicting age across all time spans, all different cellular tissues, no matter what. Dr. Horvath was very, very accurate. But Dr. Hannam's was a little bit at different, I should say extreme bit less predictive. And one of the reasons that happened is because, you know, as we age, the amount of senescent T cells in our body goes up and the amount of naive T cells goes down. And the same with the plasma B cells, they also decrease. And so whenever we're measuring the blood, we're not just measuring DNA from one cell type. We're using a bunch of different cell types. We're using leukocytes, endothelial cells, free circulating DNA. And so the cells which are represented in that blood sample change as we age. And Dr. Horvath's pattern and his algorithm sort of controlled for this and didn't really. Immunosence was not part of his algorithm. Well, it was part of Dr. Hannam's algorithm. And as a result, Dr. Horvath is a little bit more accurate across all age groups. But Dr. Hannam's being tied to the immune system was a little bit more predictive of different measures like time toward death.

So knowing that, you know, when we look at, because I checked into some of these studies that are looking at these popular aging clocks that you're referring to, the one by Horvath and the one by Hannom, and it looks like about 40% of the determination of your aging rate is due to factors that you can't necessarily control. Again, unless you were to get something like CRISPR gene editing done, like your DNA, or like, you know, how your parents lived or what your mom did when she was pregnant with you. But that, of course, means, you know, based on my rough math from sixth grade, that that's about 60% that we could change, that we could affect. Now, as you guys are getting all these tests in and you're looking at all of the different things that can affect aging, what I want to know what's most important to me are the things that seem to most significantly affect either a decrease in the rate at which I'm aging from an epigenetic standpoint, or an increase in that rate. So now I want to get into some of the more practical applications here. So as you guys are getting all thesethese tests in and running all these results, what are you finding as being the key factors that I'd actually like to start with, with acceleration of aging, what are you finding are the things that seem to really influence biological age deleteriously?

Well, most of that is common sense. And you know this very well. You know, it's lifestyle factors that will definitely accelerate aging, such as smoking, not exercising, eating, a poor, poor diet. All of those things have been well documented in accelerated aging and the epigenetic clocks. You know, an interesting thing that's relevant to you especially is that in Horvath's original clock, people who were, like, really aggressive exercisers showed extreme aging. I had Ironman triathletes that would come in and they would be 10 years older by their clock age than their actual chronologic age. And it was very consistent with the people who were really heavy exercisers and I think that they've made some corrections on the, on the more recent clocks. Do you know about that, Ryan?

Definitely. In terms of its effect on exercise, I don't know that that's been validated with some of the newer clocks, but they have definitely made some changes to some of those new clocks to be better predictive value across a number of things.

Yeah, but one thing I'm curious about is we know that there's this acute response and an increase in crp, a well known marker of inflammation. And I know that CRP is a significant increase, explanatory biomarker of epigenetic age acceleration. But is there a difference between something like the acute CRP that might have been present in a blood sample that was set in because someone had lifted weights the day prior, and chronic inflammation and elevated CRP that might be due to other factors such as poor sleep, high alcohol intake, stress, et cetera. Has that been differentiated at all? And I'm just curious because, because you don't see a lot of say, ultra endurance athletes or extreme exercisers actually dying early, except for a few kind of fringe cases like bodybuilders dying of cardiomegaly or left ventricular hypertrophy, or some people dying of electrical abnormalities due to extremely excessive endurance exercise combined with mineral depletion and dehydration. I mean, I'm just curious how much you're looking at, at whether or not that CRP is just an acute elevation because someone had exercised a couple of days prior.

That's the nature of epigenetics too. I mean, you know, exercise is probably one of the most damaging things if you looked at it from an isolated standpoint, saying, you know, after a bout of exercise you have some of the highest levels of free radical generation, you have muscle tears, you have micro traumas in the joints. But that we know over time, that degree of intermittent stress that occurs in six months, you'll change the expression of a third of your genome. I mean, 7,000 genes change expression over a six month period of exercising on a regular basis. So a lot of it is adaptation. And I can see what you're saying, if somebody is, if they're deconditioned and they go out and exercise, you're going to definitely see some adaptation abnormalities in that. And if you test someone right after a bout of exercise, if you're looking at blood work, you're going to see changes in that. But I don't think you're going to quite see that from an epigenetic standpoint, in that in those early phases, it takes time for the epigenetics to really set in based on the body adapting to a new emergent trait.

Yeah, I hear a lot of talk and, you know, kind of like the paleo and ancestral health and kind of like primal movement community, that extreme exercising is going to shorten lifespan. And have seen some blood markers and epigenetic markers or biological age markers that seem to suggest that might be the case. But then if you look at actual epidemiological data, it seems it's very difficult to find extremely fit people who are dropping dead earlier, or large populations of marathoners or ultramarathoners or crossfitters or anything like that in long term studies who are actually dying early. So that's a tricky one. But there are some other things I think, that fly under the radar. I mean, Dan, you said a lot of this stuff's obvious, and I think it is like alcohol intake or smoking or sedentary lifestyle, for example. But in terms of prediction of lifespan, what about something like socioeconomic status or college education? Have you guys at looked at things like that?

Yeah, and one important thing to note is that most of the data coming out of here tend to be epidemiological data. Right. Not interventional data where you do it before and after, after someone, for instance, has some type of intervention like exercise. And so with that being said, there's a ton of correlation between education. Higher education means you traditionally will have a lower aging rate. Beyond that, high stress or stress, particularly as a child, with psychological stress or any type of abuse can severely increase your epigenetic aging rate as well. And so there are a lot of different things that affect that, and quite a few studies now that are coming out looking at these epidemiological markers. For instance, alcohol, high alcohol intake might be negatively associated, but it's actually been found that if you have at least one drink a month, you actually can positively affect your epigenetic aging rate.

One drink a month, that's one precious cocktail.

Exactly. Well, it looks like the trend continues to once a week and once daily as well. As long as you're considering just beer and wine.

Yeah. I don't want to live forever if I can only have one drink a month.

I'll put it that way.

Okay, now, in terms of sex differences, what about male versus female?

Yeah, just as you would probably expect, due to the fact that females have longer lifespans, females tend to age at a lower rate. And. And obviously again, that makes sense, but looking at these cellular markers, it is sort of confirmed across all different types of populations.

How about ancestry when it comes to ethnicity, have you guys been looking at any of that data? Like, do we know that African Americans might live shorter or longer versus Hispanics versus Caucasians? I mean, is there any data about that?

Actually, the African Americans have longer intrinsic age in general. Interesting. That they found.

And that would be due to a lower or I guess that would be a higher methylation age.

No, their intrinsic aging is slower. And so it's basically their immunosenescence that we're looking at with that. So they're actually aging much slower.

Okay, so African Americans have indications of a significantly younger immune system age than Caucasians, for example. That's interesting.

And there's actually been some really cool information with even indigenous populations as well, where we see that they traditionally actually age a little bit quicker. And most likely that's probably due to the amount of exposures they have to infectious disease agents and these chronic inflammatory loads. But even for instance, Hispanics traditionally have a significant older extrinsic epigenetic age than Caucasians, and as a result also some fewer naive CD4 cells. And so there is definitely a correlation between all different types of ethnicities.

Yeah, you know, obviously right now, because we're kind of in the middle of hoping towards the end of, but staying in the middle of this COVID pandemic. And air pollution has come up multiple times during this, this period as being something that could predispose one to higher susceptibility to an infection when it comes to air pollutants. I've been pretty shocked at what I've seen as, as far as the significant rate of aging and risk of death in response to pollution and particulates that are pretty much all around us. Have you guys seen this in the epigenetics testing you've been doing?

Absolutely. And in the truage report, we even addressed that in particular, because these measurements of air pollutant rates, usually defined as these particular matter counts, are directly associated with increased epigenetic aging.

Yeah, I'd like to see some of these lesser known variables that people are surrounded by, you know, including non native emf, including things like the nature of the water that folks might be drinking, even things like biological versus non biological lighting. Like, there's a lot of things in our personal environment throughout the day that can be changed. Like it's easy to replace the light bulbs in your home or office with biological led, get a HEPA air filter, get a water filtration system. But I would suspect that what I've seen with air pollutants as far as vastly accelerating this, this clock that we're talking about, I imagine we'd see some similar things. I don't, I don't know if there have been many studies done on things like water quality or exposure to EMF or even, you know, biological versus non biological light. But those would be other areas that I think would be super interesting to look at.

I'm not overly familiar with it in regards to the, to the CpG methylations, but you know, we've done a lot of research in environmental design. My wife's doing a lot of the work in the military with that as well. And there are huge impacts on gene expressions with a lot of the environmental pollutants. Not, not only the air, but the light, all of that is playing a role. So, you know, I think you're seeing a big, big jump in people getting into environmental design and neuro aesthetics of home and workspaces.

Interesting. Now, it's no secret that calorie restriction and some semblance of fasting in multiple studies in yeast, fruit flies, worms and beyond has been shown to increase biological age. But when it comes to the nature of the diet itself, the composition of the diet itself, have certain diets, like whether it's a paleo or a carnivore or a vegan or a Mediterranean or any variant thereof been looked at as far as how it might influence in a large population the rate of epigenetic aging? Is there a certain diet that seems to painting with a pretty broad brush seem to be a good idea when it comes to epigenetics particularly, I mean.

You have to look at context with each individual. I mean, what's their ancestry, you know, what are their gene expressions really? Have they throughout their ancestral time been optimized for certain dietary patterns? Like, you know, you have an Inuit, Eskimo who's grown up in that, in that environment and you know, they eat a high saturated fat diet, they're going to thrive on that. But you put them on a high starch diet, they're going to get heart disease. And the opposite can occur with people from an ancestry where they were predominantly starch based and not much fat. So you can have impacts from what your past life experiences are coded in your genes, and your genes are optimized for a particular pattern that we look at for genetics. But the Mediterranean diet, not only is it kind of more of a really a universal quality diet, but aspects of the Mediterranean diet, especially with the extra virgin olive oil, the omega 3s that you get from fish, the high phytonutrient content, those all have huge epigenetic impacts. I mean, you will shift gene expressions just from the bioactive nutrients in that particular diet, for sure.

Yeah, I would tend to agree with you on that for a couple of reasons. I know that they. They are running that new age project right now that's looking at dietary strategies for aging. And this is a European study, and I believe they found really promising effects of the Mediterranean diet, specifically in regards to slowing the aging process in a pretty diverse array of individuals. And I think that people who are of Polish origin, for example, responded particularly favorably to a diet that used to, you might say, at first glance, would be the diet more appropriate to someone from a more Mediterranean versus a northern European region like that. But then the reason, I think that the Mediterranean diet, you know, people often ask me this, they're like, well, if you were just gonna choose one diet, what would you choose? And I say, well, it's tough. You know, there's, as you've just pointed out, Dan, a great deal of genetic individuality. But if I had to choose and you sent me to a desert island with one diet, it'd be the Mediterranean diet. And it's probably just because of this whole idea that the Fertile Crescent was likely where the first settled agricultural communities in the Middle east and Mediterranean kind of started, and many humans kind of blossomed out from there. And so perhaps just from this original place where the majority of humankind must have dwelt at one time, we could say that the diet folks were accustomed to at that time is still kind of programmed into our genesis. Is that kind of a logical thought pattern?

It really is. I mean, you know, you can modify the Mediterranean diet very well, too. I mean, the ketogenic Mediterranean diet is one of the healthiest diets from the data that I have seen. So, you know, people who want to be more ketogenic, they can do that with kind of a twist on that, on that Mediterranean. So there's a lot of variety that you can. You can shift around within the room realm of Mediterranean diet per se.

Yeah, yeah, I'm currently, you know, I follow some semblance of that. Not really a ketogenic Mediterranean diet, more of a low carb Mediterranean diet. I think for athletes and very active people. Something like a Mediterranean diet with some carbohydrate refeeds, where you're in and out of ketosis, seems to be pretty supportive to activity, to endocrine function, to thyroid function, etc, But I tend to agree that carbohydrate mitigation in conjunction with a Mediterranean approach just seems to, seems to work so well for so many people, particularly when it comes to just, you know, decreased overall risk of mortality. Now, you know, we, we've talked about some things that could of course, hold you back from an aging standpoint. Things like, you know, the activities of your parents, air pollutants, environmental factors, you know, alcohol, smoking, etc. But what about the things that you guys have found, you know, aside from dietary factors, to kind of increase biological age in a significant manner? Like, you know, we hear all sorts of anti aging compounds being bandied about like resveratrol and NAD and a whole host of other things. But have you guys found in the research that you've done in putting together this test certain compounds are just kind of like hell yeses when it comes to slowing down the aging clock?

You can't, you can't really say anything definitively right now. I mean, Metformin for sure, I, I don't have any doubts about the ability of metformin to really slow or reverse that clock. But the TRMM trial with Horvath and Fahey, I mean, that was, and the interventions they used in that, I have to say they're like minor league interventions.

That one was metformin, growth hormone and dhea, right?

Yeah, and vitamin D. I mean, they're basics. But we have, I mean, it is exploding right now in what we have available. I mean, we've got a whole host of senolytics that are either prescription or supplement based and they're not very expensive. We even have a peptide. This is senolytic. So we're trialing a lot of these right now with like dasatinib, quercetin, fisetin, foxo4dri, all of these that have the ability to substantially reduce senescent level load. And you're not going to substantially have an impact on age reversal until we address, you know, really all the hallmarks of aging. I mean, from mitochondrial health to cellular sensing to senescent cells. All of these things are going to have to be addressed at some point. And, and we're gradually accumulating that. I mean, I was talking on age reversal at Paleo FX six years ago, and I was looking back at my talk because I was supposed to talk again this year on the topic. And everything I talked about in that was theoretical at that point. And now we're actually seeing these reaching clinical practice now. So it's really exciting to see some of these new interventions I mean, even like Apidilon, from a peptide standpoint, cerebralysin, all of these really cool new interventions and they're interacting of one people. I mean, you're not going to have the longitudinal study. We'll all be dead by the time the longitudinal studies come out, tell us whether it worked or not. So hopefully with epigenetic age, we can do these interventions and see in a one year period that it's actually having that biological impact. I love epigenetic age as a new marker for this.

Exactly. And we're working together to do a lot of different research trials on interventions, doing things like NAD, doing things like Thymos and Alpha 1. One of the reasons they chose growth hormone in that Fahey trial was because of thymic rejuvenation and increasing the ability of the thymus and decreasing aminosenescence with age and things like thymosin. Alpha 1 would make a lot of sense for that investigation as well. And one of the things I should note is that the Fahey trial, the TRIM trial, is really the only interventional trial that's been published at this point. And it was sort of a proof of concept, only nine Pat. But they're actually expanding that now, I think to over 100 patients in the TRIM X trial and are actively recruiting patients. And so hopefully we'll know very, very soon a little bit more about a lot of those interventions.

Yeah, I'm a little bit kind of on the fence about completely shutting down cellular senescence with something like Denisitib or high dose Quercetin, for example, at younger ages. Just because, I mean, we know that senescent cells play a crucial role in mediating tissue development, development in the formation of the embryo, and also tissue regeneration and wound repair later in life. You know, it's kind of when the, when the so called zombie cells of senescence accumulate with age, that's when you get a lot of inflammation and tissue dysfunction. But I think there's gotta be a sweet spot at which you would want to really begin to focus on limiting cellular senescence. And so, you know, someone sent me a bunch of Denastib and Kercel and they said, oh, this is going to be the greatest anti aging compound you could take. Ben. It's honestly, it's in my refrigerator. I haven't touched it because I have yet to be convinced that at my age, at 38, I want to significantly limit cellular senescence. Do you guys have any thoughts on kind of the sweet spot about senescence?

I think it's really a question of your individual context. And your aging process. I mean, we're really good with clearing senescent cells when we're younger. And as we get older, we become less and less capable of doing that. I think it's a big part of the programming of aging in the DNA. And you also don't want to be overly aggressive in somebody who comes in who's 75 years old and give them a really aggressive senolytic approach because, I mean, that's going to create a lot of inflammation in the body. So you've got to kind of temper that with each individual. And you also also don't want to run senolytics in a continuous run. There's timing to the way we do this with age rejuvenation. I mean, you've got to basically go in and tear the system down and then build it back up. So, you know, you come in and you hit them with the senolytics and start with the thymic rejuvenation at that point. Then you start into the NAD and then potentially exosomes, growth hormone secretagogues at that point. I mean, it's a, it's kind of a. Right now we're, we're doing a lot of n of 1s with this, but you know, it's there. You got to use some common sense in the way you're approaching it. But, you know, one course of dasatinib in a, you know, broken out over a three week period with two days each week, and you can get a substantial 30% reduction in senescent load in many cell types with that. And that is going to be huge. I mean, our stem cells are basically poisoned by the senescent cells. And once you start depleting that load of them, those stem cells start to kick back in and the other cells start functioning well. So senescence is a huge, huge area. But, you know, we can't attack that without looking at how do we boost nad, how do we boost telomerase activity. Got to look at the entire picture of the individual and take that approach. But I agree with you. I mean, at 38 and your health, maybe some minor senolytics like the quercetin and the fisetin, those might be beneficial for you at this point, but hitting the dasatinib or the Foxo 4 Dri is probably not advisable at this age.

You can also consider it though, as a way to promote quiescence or to prevent the GERO conversion to senescence. And so that's where even things like rapamycin and MTOR can prevent that irreversible senescence step. So you can think of it more as a preventative strategy with other compounds as well as a way to even things like these growth hormone secretagogues as ways to keep these cells from going senescent in the first place.

Okay, got it. Now let's say that and just a fun thought exercise here that. But either of you guys tested, I'd love to hear each of your responses. And your results came back and your biological age was higher than what you would like to see. And you're already eating healthy, you're avoiding excess alcohol, you're having your one cocktail a month, you're not smoking, you're taking care of relationships, lifestyle, stress. Maybe you have yoga or meditation. Practice this. What are a few of either the say, biohacks or lesser known supplements or anything else that you might do that you would consider to be hell yeses for you? Armed with the knowledge that each of you guys have about how you would improve that biological age?

I can tell you from personal experience, I mean when I was 39 years old, I did a telomere test and from life length and I was 10 years old, older my biologic age than my chronologic age. And you know, I was a general and vascular surgeon at that time. I slept very little. I ate pizza and drank Mountain Dew all day long. Was not overweight because my, my exercise kind of masked my, my unhealthiness. But that was an eye opener for me. And I completely shifted. I started really getting on a regular sleep schedule. I quit surgery for many reasons, but health was a big one. I started meditating, I started exercising regularly, eating a healthy diet. But I think sleep is probably one of the biggest things that we see in our clinical practice that people are, they're undervaluing the benefits that sleep can give them from so many angles of aging. It is a high priority for sure.

Okay. So that, that's not necessarily uncommon knowledge, but let's, let's dive in just a little bit more to that. Is there anything that you do for sleep that you think more people should know about who don't know about it?

Well, the biggest thing is I get on a regular sleep schedule. So I'm in bed around 9:30, 9:15 and 9:30 every night. I wake up without an alarm clock every morning around between 5 and 5:15. And I keep to that schedule as aggressively as I can. And when I'm off that schedule, you know, I monitor, I wear wearable devices all the time. I've got a whoop on one arm and a garment on the other. And I track all of this and I watch what happens. And I think this is an important thing for people to get is start learning, learning interoception, understanding the body and the wearables can kind of help teach you that process and learning. You know, you have a glass of alcohol. You know, I have, I have one glass of wine and my stress level goes through the roof for about eight hours. And you know, for my HRV tanking and that's okay to have periodically. I mean stress is not a bad thing periodically. But if I'm having that glass every night, it's, it's certainly going to have a health impact on me. You know, I take things like metformin for aging, you know, for the calorie restriction. I intermittently fast, but I don't tend to calorie restrict. But the metformin will tell my body that I'm basically calorie restricting, which is nice. So there's hacks around that as well.

Yeah, just goes to show you too the level of individual variation on this because I tend to have a drink most nights of biodynamic organic wine or like a really nice cocktail, like a really high quality bourbon or rye or gin with a splash of bitters and some lemon juice. I actually just now while we're talking, I pulled up my aura sleep scores and Tuesday was the only night this week on which I did not drink. And I've got sleep scores of 90, 89, 67, 88, 92 and 85. And the one night on which the sleep score was low was the night I did not have a drink. And part of that could also be the fact drinks have carbohydrates. Carbohydrates can assist with serotonin production, enhanced relaxation and perhaps help to fill some glycogen reserves to a certain extent. And so part of it could be that as well. But for me personally, in my own internal testing, a glass of wine or a cocktail a night actually seems to help out quite a bit. But I think one really important thing that you highlighted, Dan, was this idea that you don't go to bed at let's say 11pm every weekday and get up at 4am and then catch up on the weekends. Regular sleep cycles in my opinion are underemphasized and super, super important. Now how about you Ryan, anything you would do if you got back with a low biological age that you just think would be a non negotiable for you?

Yeah, I think that there are two Considerations for there. And unfortunately, I don't have some secret intervention that's not well known. I wish I did. But I think that one of the things to consider is where you're at and particularly what your age is, because what the data shows is that your chronological age is really correlated to how much of a difference you can make in these therapies. The one thing about this Stahy trial that was so exciting is that they reduced the epigenetic aging rate by around 2.5 years, which is a pretty crazy statistic. You know, they even looked like toward the end they were accelerating to even more age effects and even going up at a higher interval. And so the reason that's important is because, you know, for older individuals, I think that, you know, exercise, particularly aerobic exercise, has more of an impact, whereas if you're younger, I think that things like sleep, exposure to pollution, exposure to environmental toxins are a little bit better. So I think it greatly depends. The intervention depends on your age. But ultimately, I think that if you're. If you're not doing the intuitive things in one area, that should be your focus.

Yeah. Yeah. Well, you know, when it comes to this actual test, again, it was. It's pretty, pretty simple, really. You guys sent the kid to my house. I put the blood drop in the collection tube, which I think has some EDTA in it to prevent coagulation, and then shipped it back. I'm still waiting on results. I think it'll be about a week out when I get them. I will probably get them by the time I publish this podcast. So for those of you listening in, what I can do is put my actual sample results into the show notes. If you guys want to see what a sample report looks like, I'm going to put the shownotes@bengreenfieldfitness.com truediagnostic it's spelled just a U T R U diagnostic. And then if you want to get this test yourself, it's pretty simple. You go to the website, you order the test, I'll put a link in the show notes. Ryan and Dan are giving us a $50 off code on your True Age Kit. It's just Ben 50. If you go to BenGreekvilleFitness.com TrueDiagnostics you can use code BEN50 to get your test. Or actually, it's true diagnostic, not true diagnostics. And then what is it? Guys, guys. It's about three weeks to get the results from what I understand. Right?

Yeah. By the time the test is published, it should be around three weeks to get the results and they tend to be relatively comprehensive. It's a, it's a big report, but we'll address all the things we talked about.

Okay. Alright, cool. Well fellas, anything else you want to throw in? When it comes to the status of these methylation clocks, the status of age testing or anything else that while you're here, you want to share with folks, that's, that's something you want to, you want to get off your chest or you can just go have a cocktail. It's up to you.

I would just want to emphasize that, you know, don't get hung up on the, on the absolute value that you get back on that it's more about the dynamic over time. And you know, we're going to get more accurate over time as more and more samples are brought in and we compare this to other markers. But you know, if you come back in two years over your current age, it's not something for somebody to get all worried about and start panicking about something. What you want to see is what's happening with that age dynamically over time. A year from now you repeat that. And this is why I like epigenetic age over telomeres. I mean you got to wait five years really to get really actionable outcomes from telomere testing. With epigenetic age, I mean we've done some six months apart with looking at interventions and seeing how it's creating a dynamic change in that. And even at six months, we're seeing effects of lifestyle patterns that are shifting the rate of aging in the epigenetic aging.

Wow, it's impressive. Well, and you're right, I mean these are all modifiable factors. And we know that there are gin chugging, cigarette smoking grandmas in Sardinia who are living impressively long times. And so we don't necessarily want to think ourselves into via placebo effect, accelerated aging or just identify ourselves with a higher biological age. But I consider any of these tests to simply be a method of equipping and armoring yourself with the knowledge that you need to make really, really good decisions about the diet you're going to follow, supplements you're going to take, modifications you can make, etc. So I think that's where the value really lies. Alright folks, well that's what we got for you. And again, the shownotes are@BenGreenfieldFitness.com TrueDiagnostic if you want to get a test, you can get 50 bucks off with code. Ben 50 and Ryan and Dan, thanks so much for coming on the show and sharing this stuff with us, guys. And I'm pretty excited to get my results back and thumb through them and learn about how much I actually can drink and whether I need to take any NAD or resveratrol or inject peptides or any of these other. Any of these other things that we can do to live a longer time so we can make maximum impact on this planet with the life that we've been given. So thanks for coming on, fellas.

Thank you.

Yeah, thanks for having us.

All right, folks, I'm Ben Greenfield along with Dan Stickler, Ryan Smith signing out from BenGreenfieldFitness.com have an amazing week.

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