David A. Sinclair at ARDD2024: The Information Theory of Aging: where do we stand?

and can we get David Sinclair to the stage woohoo Alex Jesus Christ okay thanks okay we're on uh thank you Victorio for the uh the great presentation and the intro uh I think we're all in the mood for some Rejuvenation uh it's late in the day nice to see uh lots of friends and old colleagues and thank you to the organizers you're always very generous and kind uh and thank you for inviting me thank you all for coming uh so we have a lot to talk about today uh I'm going to uh give you an

update I was here last year and we had just published the ice paper on being able to uh test a large part of the information Theory of Aging which I will get to um that paper uh came out in cell in beginning of 2020 three uh after 13 years of work um 46 researchers uh 30-some labs and three mortalities of our co-workers along the way it was an incredible journey and I was very proud to present that to you all uh a year ago and we' also published uh a little bit earlier than that that

we could also not just in these mice Drive aging forwards by uh altering the epig genome but also as Victoria has told you we were one of the lucky groups uh I guess resting on yaka and Belmonte labs to be involved in uh the field of Rejuvenation and I want to give you an update largely on that work now where are we I said that when I came back I was going to tell you a number of things about the chemical reprogramming about where the information lies that we think for Rejuvenation uh update on clinical

trials I'm going to do a lot of that today so what is the information Theory of Aging uh it's a culmination of a lot of labs work I'm certainly not the only one to suggest that the epigenome is important um I became involved in studying the epigenome uh through Fortune good luck working with Lenny GTI in the early and mid 1990s uh on yeast aging and many of you are young so you may not remember those days but what we discovered as a team in Lenny's lab and I was a posto then uh is that

there's a set of genes called the cin named after the CTU Gene and ceru is a hisone de acelas that controls the epome S stands for silent inform regulator so information is key and of course because information was part of the genes I was studying I've been fascinated throughout my career in what the heck is information doing as a subject in aging and I became particularly fascinated with this as we learned in yeast and then in melon cells that we could see predictable changes not just in one cell type but in many different cell types

and tissues and species changes that were predictable in the epig epome used Loosely meaning hisone changes methylation changes that Victoria has just talked about we came up with a theory that's not called the information Theory of Aging called uh the more of a mouthful the relocalization of Chromatin modifiers theory that was in 2008 with Philip obor as a postto and that really was the beginning of the of this Theory it was a predecessor and what we showed in that theory in that paper that was um published was that we could disrupt and accelerate epigenetic change

in a way that mimicked aging by creating very precise non-mutagenic double stranded braks and we could see that we could drive aging forwards in cells uh and now we know even in animals to do that uh and then with the later work by our lab and of course many others now um and Bel Monte before that and Victoria was there uh with that work it was very clear that to us at least that there's there's a backup copy of youthful epigenetic information in every cell that we've looked at I'll show you evidence for that uh

but I really do believe that that's true it's for me the best explanation for how it's possible to have not just reproducible changes in the forwards of Aging Direction but in the reversal as well somehow cells know which Gene to change in reversal by how much and in what direction where that information Lies We are working actively to figure that out we think it's probably something of a of a nucleic acid material uh but that's about all uh we are sure about so far um this is published so if you want to read more about

this Theory I'm not going to spend time uh educating uh about it today but I will say that uh it's largely based this idea uh on the work of Claude Shannon who discovered and formulated the mathematical theory of communication in the 1940s um and instead of a backup copy of information which he used to build the beginnings of today's internet uh we're talking about biology here and we think that there's a backup copy that we can use through Y artifactors and increasingly chemicals to reset the cell find the original information that's somehow laid down when

we were very young in cells and can even last decades we as a we're reprogramming cancer Souls we think back to their original uh state in the body and that's cell line has been out of the Body for 50 plus 50 plus years so 50 years is a long time to store epigenetic information um and as I mentioned we think that cellular damage such as double standard breaks and now we know nerve Crush um and others like vom gladish have shown that even pregnancy is a stress on the body that can accelerate aging there are

a lot of things that cause these what are now referred to as Epi mutations uh and this is the RCM hypothesis which I just talked about the idea that that the real ization of Chromatin modifiers such as the cin and of course others like HDX um are involved in this process prc2 Victoria just mentioned we we can look at our data and also find very strong associations with the prc2 complex so it seems to be very key to this process and Rejuvenation um so here's here's the summary of this talk the update is that we're

doing a lot of things we've gone not just deep but now we're going wide uh We've almost got too much to study uh and we're too excited but what we're doing um is you know everybody's doing artificial intelligence but what we're excited about is that we can find small molecules and Screen them virtually and process those very quickly um we also do reprogramming of cancer cells as I'll show you uh we're doing a lot of epigenomics where is this backup copy trying to find out how that all works at this molecular level can we reverse

cellular inessence right now I think most people would say that's impossible possible um I can tell you that it's not um is it safe I'm not sure yet but we're working on it lots of human organoids CU mice are not sufficient uh and we do a lot of work on inflammation as well which I'm not going to touch on today um I'm also not going to touch on the human clinical trial preparation Jerry uh mcglothan was here and as CEO of Life biosciences a company that was spun partly out of my lab he talked about

that uh and he presented great data on uh the beginnings of primate work to to restore visual function to Blinded primates and hopefully next year if all goes well or if you know maybe next year the year after we'll see but human studies are are definitely being lined up to test these Technologies first in the eye and then beyond uh this is the system that we use in mice monkeys and in hopefully in humans uh it's an inducible system for safety reasons we use uh AOC as the major inducer and it works really well and

what we found is that we can turn on this system for a few weeks uh sometimes for over a year in an animal it's very safe the reason that we think it's particularly safe uh is in part because it's not getting to the susceptible cells as much such as the gut lining but also because we're not using cmech which um is an oncogene and we think that's important so we're using OS and K Y Arch factors not OS km um and uh and that's what we've been working with ever since we published this was work

that we did publish so I won't go too much in detail but to highlight something that you may not have thought about as much as me and that is that the Rejuvenation of an optic nerve or a retina and restoration of vision in an old mouse or a mouse with glaucoma is associated with not just changes to the DNA methylation but gene expression changes that are uncannily similar to a young cell and this shows this if if those these are genes that go up with OS treatment viral treatment and down and if there was no

change these dots which represent each Gene's expression would be Flatline it would be no change like this in the horizontal and if there was a it was accelerating aging which obviously it isn't it would be from the lower left to the top right but what you're seeing is the opposite you're seeing that in general genes that when we when the cells were young that were down that went up with aging go back down by this by pretty much the amount that they went that they change and vice versa so that's why one of the bits

of evidence that I feel increasingly confident that there is a Mark or a nucleic acid or something on or in or near the genome that allows these yamanaka factors to reset really accurately sometimes as I mentioned decades earlier um I mentioned sence uh so you know we all stay sence cells for beta galac todas which you can see here uh what you may not know is that using OS and as I'll show you chemical cocktails that mimic these factors uh we can pretty readily reverse a lot of the gene signatures of senescent cells and even

get them to grow again in a healthy way now we don't know in Vivo if this is safe but in vitro it's not that difficult we have checked to see if they oncogenic are they expressing genes that might predict tumor Genesis and we don't see any of that so we think that what we're doing is restoring the ability of cells to regain their identity having lost their identity during aging and this is uh largely the work of junyang who's now a postdoc in Korea uh as well as Chris Petty who's here at the conference um

and Thomas uh McDougall and these are gene expression patterns that you can see heat maps of three separate experiments three different cell lines uh I mentioned Ai and you I'm inspired by all the work that Alex does we're we're primitive compared to him but what we've been able to do is use AI to our benefit and one of the things we're doing Thomas is doing is to use AI to very rapidly detect young middle and old cells from humans or mice and you can look at cells or I can look at cells I cannot tell

the difference a young cell looks like an old cell a microb a little slower but you apply AI the visual system and they and train it just for 10 minutes and it can pick out a young middle-aged and old it can even pick out how sence was was caused uh or uh or how many passages it's been through and this is a very useful method now because we can screen not through dozens of chemical cocktails but potentially millions of chemical cocktails and it's really shown how we do it here I'll just be brief we're looking

for genes and chemical libraries using Robotics and we take cells from different AG people 20s '90s um we can make them senescent and Thomas Paints the cells we used to use just one Dy uh to look at nuclear localization but now we use a variety of cell paints and that gives us beautiful resolution uh and what comes out the other end is very very accurate uh it's so easy now to tell if you've rejuvenated cells in high throughput and we've now got chemical cocktails and even single chemicals that can rejuvenate cells human cells Mouse cells

different cell types that were testing on organoids tissues and animals we've applied it for for instance to human brain organoids little little organiz that we grow in the dish we can give them Alzheimer's mutations and they develop signs of dementia in the dish we can measure the electrical activity we can rejuvenate that we can get the electrical activity back again uh in those organoids using the genes and chemicals that we're applying and even gene expression patterns go back in those little organ oids to a more youthful state so we're excited about testing these chemical cocktails

as well in disease models such as the eye um and we're also going to be testing these on animals this is an example of some of the data that Thomas has recently generated where you can see a big difference in uh the ability to detect nonsenescent versus ccent cells the blue versus orange but you can also see that it depends on how you cause cellular inessence most people just or at least you know if you're if you're not really into it you just do be galaco cyto staining and they're all blue but these programs can

tell the difference between how they got blue and uh this is these are cells here that are treated with an epigenetic inhibitor GSK 126 which inhibits the prc2 complex that Victoria mentioned that's an excellent model for aging we think of genetic aging um and there are other types of sencence such as dox rubison which creates genomic stress or genotoxic stress and we can see the difference so we've come along way um I'll be able to tell you a little bit more about those chemicals in a minute uh we've gone beyond the brain last time I

was here we're talking about cerebral organoids now we're doing hair organoids uh we're looking at being able to accelerate the age of these organoids because you might appreciate that because they come from Human stem cells they're age zero they're very hard to make old but we can use our ice system like we did with those mice and make them epigenetic older genetically older and we're looking to see if we can make this the skin old and the hair old and then rejuvenate you can actually put these organizes in the back of a mouse and grow

human hair it's really quite grotesque but interesting um and we're also doing um we're doing inner ear as well these are inner hair cells on the left uh these are some of the organoids this is worked by Shen ta who's now at the bone Institute in San Diego and uh he was overexpressing in a very uh neuronal specific manner in hippocampus OSN K using viral delivery as you can see here he stained it in the hippocampus and what he and now others have shown is that we can rejuvenate the brain and get the ability to

learn again in Old mice and he also has uh I think unique data showing that he can recover uh learning in memory and even lost memories in Alzheimer's mice models what I thought I'd point out that's interesting is some of the genes that are expressed we're seeing changes in uh the EMT transition so this is in uh the the EMT transition that's involved in embryonic development being playing a role in Rejuvenation which is pretty interesting uh but also what's exciting is some of these genes here are very long non-coding rnas that may be involved perhaps

in required we testing uh in this Rejuvenation process so rnas is as Victoria has identified is I think a really interesting area to explore uh what about cancer now we had a bet in my lab that uh reprogramming would make cancer worse or better uh and you can judge yourself how you would vote I was Voting that based on the information Theory of Aging Rejuvenation would take any cell whether it's old or cancerous orogenic and restore the identity of those cells so my bet was on them returning back to normal I didn't think it would

work on every Cancer cell cell line but I'll show you an example of the results that nalot uh in the lab see what por and chai is doing she's a graduate student very talented one um oh there's the EMT data if you wanted to take a photo of the kind of uh processes that we see in the brain get a snapshot of that uh yeah so here's nart uh nlat is uh has been working on this I think she was skeptical too which is always good and uh what we know is that if we or

others overexpress OS and M transcription factors in my it'll either kill them rapidly or create teratomas so that's not going to be a therapy anytime soon but we wondered whether if we if we leave ALC the enene what happens to a tumor that's already pre-existing and we're studying lung kidney colon and melanoma this is the data from colon cancer cells this is the first cell line that nlat worked on it's not I'm not showing this because it worked only it's the really the one that she studied first and this is an inducible system you can

see it coming on on the left um and and these the stains so what happens after induction of os in this cell line these by the way these these come from the lining of the gut uh from over 50 years ago from a colon cancer patient these are endothelial cells originally uh this is a scratch essay looking at cell growth if the cancer cells grow quickly they close the gap in the scratch on the plate uh and you'll see this graph go down quickly and the only cell line that closes poorly is the one that's

expressed pressing uh OS which is shown in the green balls here similarly uh hopefully you can see this up the back the only cells that grow slowly in this video are the ones that are overexpressing os all the other controls grow like weeds so that's pretty cool but what's happening are we just making them sick or what well yes uh they go through induced programmed cell death and in studying them what we're seeing are really interesting changes in the lome in uh autophagy uh and also the morphology they are starting to resemble they at least

are reminiscent of their original cell type and one of the examples is that I'm showing you here is the nuclear are long and extended and you see these in the columnar uh end endothelial cells that they originally were again 50 years ago so that's a really interesting idea that and we have to prove it of course I'm just showing you some of the early work that reprogramming can restore the identity of cancer cells and I think this is again speculation that when cancer cells wake up to their identity they freak out and they kill themselves

that's what seems to be happening uh we're doing a lot of EP genomics trying to figure out how does reprogramming work and this is just an example of some of the data that we're getting this is by son mayber Lewis and Chris Petty in the lab and this is an example of where socks 2 ker for c one one of the serans and tatto tatto being a DNA demethylase enzyme that they're going in large part to the same places during reprogramming this is in the first four days of the of the process obviously later is

different but the first four days they seem to cooperate and it's interesting that cin are involved as well and the kind of processes that they move to are genes are involved in stress DNA repair and cell cycle again really interesting so just briefly about the small molecules remember I was going to talk about that last year was just preliminary we've come a long way these are the the chemicals that we published uh last year uh and you can see them here I'm I think some people are already trying to take these I wouldn't advise it

some of these are not exactly known to be safe but we're trying to make them safe so instead of it costing $200,000 to rejuvenate a tissue let's make it $200 that's our goal and make it safe the essay that we used to use and we still think it's useful is uh to look at whether Pro proteins and leaking out of the nucleus which happens during aging and inessence um or if they go back inside the nucleus we call that Rejuvenation and it's a good s surrogate for for that uh and this is a good example

of the data that we get these are leaky nuclei in the senescent cells and after treatment with one of the cocktails called c6f which is one of the more potent cocktails from last year uh we get almost a complete restoration of nuclear Integrity even better than the gene therapy which blows my mind this is largely the work of Chris Petty and Thomas again so where are we at we we've done enough work on these chemicals that we've now identified sets of them and even a single chemical that we're treating mice with we know already that

they're safe in Vivo and we're going for it we're going to see if we can reverse inessence in Vivo s inessence and hopefully lifespan will be improved uh and perhaps Health along with it uh and that'll probably be for next year but that's where we're at so what I want to emphasize is that what seems to be apparent from work from other labs like voros and uh Juan colus Bel Mon's lab any others Tom Rano we are seeing that reprogramming can restore identity of cells from many years before allowing them to behave like they were

young again quite literally because they are are young again and this is an example of that shown graphically where over time during aging cells lose their identity becoming this red ball and wton would be proud We Now seem to be able to push the balls back up back to where they came from uh even if they're cancer cells in some cases uh so I will end here happy to take questions and there's a lot of people I mentioned along the way uh yuchen GAA I didn't mention I should have he did a lot of the

organized work and he's charging ahead trying to restore hair growth and gray hair that'll be fun um and minan talks about inflammation but I didn't have time to talk about her work today and the many many collaborators over the years and I want to just point out um in particular vams helped a lot with clocks ver has helped out a lot with the RCM work and John with transposon work and John's here as well so thank you very much want to thank the sponsors and for inviting me back again thank you thank you so much

thank you David um we have questions excellent uh presentation and I have a question about a small molecules do you think that your therapies can be eventually done with a small molecules oh well that's the goal I from what I've seen I would say probably I would I haven't seen let's put it this way I haven't seen anything that says that they're dangerous and that they don't work in human organoids or in cell culture I don't know yet uh really I don't know yet if they work in Vivo but I I don't know why they

wouldn't okay we're going to be able to bathe all the cells in the body with these chemicals uh which is different than gene therapy which is very hard to deliver and we'll see what happens I think it it should work question is how safe is it and that's obviously on our minds um I will ask a quick question there's a couple question in the back but while the microphone is going there then I will ask a question so they regarding the um the substance the secret Source whatever it is that makes a cell uh identify

as that particular cell I was wondering have you thought about like secondary DNA structures or something that's already in the genome that could affect DNA methylation in the area yeah we have thought about that yeah we we think there's some structures that are laid down like that that there's a whole different set of proteins that I that recognize those structures it it I think it'll be similar to crisper how that works there's a barcode at the gene that tells it what it used to be by how much to change it and in what direction yeah

fascinating um hi David very very nice talk sorry two quick questions the first one is on the cellular Essence reprogramming are we talking like enene IND in essence because this involves complex chromatin rewirings or are we talking like replicative in essence which I can see why that would be easier to reprogram uh We've reversed the three main types that I talked about um replicative uh chemical induced uh and uh and genotoxic stress um unless Chris is going to jump out and correct me I don't believe we've tried oncogenic uh sence gen cause inessence but I

agree with you that that may be harder to uh reverse okay thank you and sorry one more question one question sorry because we we are running out of time so we we have to get more people chances here vorio nice nice talk David I'm here okay nice talk lovely thank you um it's I'm very excited about what you seen on the on the cancer cells uh and the for two reasons well obviously maybe one day we could think about partial reprogramming as an anti-cancer therapy number one but number two I think also it answers the

question that keep being asked that probably you also keep being asked like what happens if by chance you target a precancer cells as you do the Rejuvenation you know in the body and I I think it's kind of well new data needs to come but we can even start thinking about the safety of the approach also as a way that you know even if cancer cells get targeted maybe there is a chance that actually those are going to go into through apoptosis die off and so it's going to it's it's a higher degree of safety

I I would say would you agree yes I I agree and that's one of the reasons we did the experiment besides testing the theory is that we want to know if it makes cancer cells more active and we're getting the opposite results so far very excited last question um yeah thanks so much for the for the great talk um I was curious about the efficiency of the reprogramming with okm because with okm reprogramming for ipscs um some Reports say that's quite low 1% or so um and I guess that varies but I'm guess curious about

with the partial reprogramming what the efficiency is and if there's ways to increase that uh it's really really efficient it's not like uh generating ipscs would you agree uh where's Victoria anyway so we we when we did the the retina with Bruce canders and this was the nature paper in 2020 uh we could stain for OS expression in the retina in the ganglial cells in the neurons and stain for the restoration of cell identity using a marker for rgc's and every time the neuron got the OS and it was expressed that marker came on so

the delivery is the hard part it's not the the restoration of Youth in the cells which contrast with ipscs all right thank you so much David we will move on we have a panel after this so really appreciate it thank [Music] you we will have uh let's have a 10-minute break come back at 8:30 for the farming panel it's going to be really exciting and after that we go to the bar