Fight Aging!

As I've mentioned previously, reprogramming is the area which excites me the most after snolytics. I think this field is going to explode in the next few years, as evinced by the following new podcast with David Sinclair, where he provides new details about reprogramming work undertaken in his lab in the last year (amongst other topics):

https://www.youtube.com/watch?v=J736mfy7KEg&feature=youtu.be

@Chris
Reprogramming itself call be done more or less easily, after so many years of research. The problem is to instigate the right growth and differentiating process. We want to induce pluripotency but avoid tumors and malformations.

So in a way the first generation stem cells treatments failed to differentiate the implanted stem cells but that was a good thing since they were safe. I remember if one case a few years ago where a woman got injection in her eyelids and mini bones started to form. Apparently, that treatment protocol managed to preserve the stem cells, so they didn't die but started differentiating and forming tissues. Unfortunately, wrong toe of tissues in the wrong place...

Hi Cuberat - yes, I agree with you. I should have been more specific when mentioning reprogramming. I meant transient or partial reprogramming, which rejuvenates the cell/organism whilst avoiding the formation of teratomas or other cancerous formations.

Today, the idea of using Conditioned Medium from Human Mesenchymal Stromal Cells is encouraging, but unlike the authors of this article (https://doi.org/10.3390/ijms20071656), the Mesenchymal Stromal Cells must be previously irradiated as in the article (http://europepmc.org/articles/pmc5745544) to activate the alarm signaling for regeneration. Also a source of controlled prolonged release of ROCK inhibitor [http://europepmc.org/articles/PMC5551544] must be added. Vector of Δ133p53α (a natural p53 isoform), which induces hTERT expression and telomerase activity [http://europepmc.org/articles/PMC6030220], can be used instead of irradiated cells. See more details in https://www.academia.edu/38872702/Some_suggestions_on_the_use_of_conditional_cell_reprogramming_technology_for_tissue_rejuvenation_in_situ

As Reason repeatedly points out, it is the signalling of these induced stem cells that is critical, and this also lays behind the cancer risk. So long as the signalling is not too strong, which means the number of cells is not too high, no terratomas will form. Therefore iPSCs can be injected into circulation and form no terratomas. Plus because these cells are small and can cross various tissue barriers without impediment, they can locate to a site of injury and differentiate, effecting repairs (unlike adult stem cells, which are useful for repair in far more limited scenarios). The evidence for what I am saying is here: https://www.ncbi.nlm.nih.gov/pubmed/30662568

Reason implies that the cancer risk comes from the fact some cells are still 'damaged' in an obscure way that has not been fixed by epigenetic reprogramming. However, as I explain above, the cancer risk comes when the reprogrammed cells override the signalling of the tissue in which they are placed, not from 'damage'.

Either injecting iPSCs or as Dzhagarov points out, a conditional reprogramming approach, is going to revolutionise health care and will form one of the main planks of a rejuvenation therapy in the next 20 years.

teratoma risk is also heavily dependent on the exposure to reprogramming factors, you want a partial reprogram enough to erase the aging markers but not enough to loose cell identity. This is ~4 days in many tissue types but it depends on the target cell. It is perfectly plausible and already demonstrated to be able to separate the aging marker reset portion from the cell memory portion of the process as the markers are reset earlier in the process. Partial reprogramming has a huge amount of potential given that epigenetic alterations are a primary cause of aging.

Teratoma formation is a morphodynamics issue

This has been known since the 1970's and the seminal work by Dr. Beatrice Mintz et. al in Philadelphia (https://en.wikipedia.org/wiki/Beatrice_Mintz) in embryos

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC433040/

Rejuvenation with any of these cell types must take into account the morphodynamic processes occurring in the ADULT human body

The organisms on this planet that have the ability to reliably turn back "biologic age" in some (or all) of their bodies, using some form of reprogramming, possess two important inherent attributes. These are:

1) The ability to re-establish the "embryogenic" potential of their cells / genomes,

AND,

2) The simultaneous ability to re-establish the "morpho-dynamic" architecture of their tissues / organs / limbs / body segments, once such embryogenic potential is re-established.

The last 100 years of regenerative biology research has shown that one, without the other, is an incomplete path towards rejuvenation.

@ Steve - everyone is focussing on partial reprogramming because we supposedly want to maintain somatic cell identity. But what I'm saying is that this may not be necessary. An IV drip of iPSCs may be preferable. Somatic signalling will dominate the (relatively small number of) added cells and they will differentiate appropriately, satisfying the "architecture of tissues" as Ira says, rather than inappropriately, as with terratomas.

This is analogous to the in vivo work on age accelerated mice, where they induced reprogramming for a limited time, rather than on a limited number of (ex vivo) cells. Obviously in vivo is higher risk. Given ex vivo will be affordable (everyone in the cancer immunotherapy space is working on it), it seems the more pragmatic option.

We will see. I am optimistic we can have our cake and eat it, it's all about exposure time.

Hi Ira! Just a 2 cents.

Do you think that epireprogrammed cells (via yamanaka) have this possibility of remorpho-dynamically remaking architecture of tissues and organs, but in very aged adult body/like an elder of 90+ years old, specifically ?

When I look at the litterature, it says that by this age, the damage accumulated is just 'highly consequential' (or too consequential?) to be able 'to revert' to young 'architecture' of tissues/organs..
meaning, it has come to stocastich 'irreversibility' of current architecture, because it's so late (90 years later), the person's body is just not 'reversible enough' aymore....just like a epigenome that cannot bounce back by itself (if it using no epireprogramming but 'regular epiaging').

That worries me because it would mean (just guessing) that there is a irreversibility point to epiprogramming - that the epigage of cell can be reversed zero but it does not mean Reversal of Architecture of tissues/organs, neither. For that to happen, the environment - still young enough - must be so to 'allow that possibilty', as you said, like say..in a eternal hydra jellyfish which has telomerase germ cell line in its tissue and can revert its 'adult/mature' state to a immature child state (by its own self-epiReprogramming program after sexual reproduction).

Thus, there could be a 'cut off/tipping' point/'PONR (point of no return)'..if you past this age, the likeliness of reversal of organs/tissues to morphogenic architecture is not feasible anymore because of so many 'changes'/epialterations accumulated at late age and in the architecture. The damages, so late, may then be too consequential to allow rearchitecturing.

It's oftently what happens with very advanced diseases, it seems like irreversible; I want to believe that is reversible, and I hope it is. When I look at hydras jellyfish they do it...but do not go to point PONR...they don't age so Late that it's 'TOO late' for it...

and this whole epigenetic reprogramming is confirming that - you have a 'window' or 'it's too late'...you get cancer/Teratoma formation...I'm starting to think that, in a very aged body, it'S the same thing - too late/irreversible. I hope I'm wrong though, in the litterature, there is basically 0 animal that can reach imm*rtal state by epireprogramming - at a Very Late age/old state...the trick is they keep 'old enough/young enough' to make it still viable/feasible/possible. Too late/Too old..is too late/too old. Let's keep fingers crossed and hope that we can Overcome the 'cumul' curve of aging and rearchitecture them to a young state..it may take long, but does not matter...as long as it takes...to make it happen and Reverse an very old body to a young body. But, not holding breath. Rejuvenation/age reversal was, it seems, something is a 'time limit' to it and can't wait 'forever' to do it (there is such a thing a too late).

Just a 2 cents.

PS: I see two possible main outcomes of this

1- In very late aged (above 90) = therapeutic/health promoting, the person may get a extra few years (10 or so), but they will die around 120 MLSP. This will make healthy aging, not reversal of aging.

2- In younger age (below 60) = longevity creating, it could very possibly make humans go above MLSP and then, the possibility of LEV would be concrete and not so remote anymore, the trick is making that epireprogramming continuously back to epiage of 0, but staying in that 'no later than 60 or so' in current architecture of organs/tissues - to continuously allow repeated reprogramming (that is something not spoken oftenyl Repeatability of Reprogramming - Re - Re RE......forever in loop di loop, how many times can we do it until death happens, if we reverse, it could be eternally, but nothing says that that is a possbility if we cannot keep organ architecture intact to that of someone young enough, to still that possibility or 'repetaed' reverser re-re-re-..reepireprogramming and Re-re-re-...rearchitecturing). I await that studies do it ****Repeateadly**, not once and 'called it a day', that's mostly not very 'telling'/just surface reversal...do it Repeated/continuously/on and on, and then will see how viable this is over multiple repeated times (like will we get teratoma formation/cancer?/it is still architecturally 'sound'), over a long time; this would confirm us the True repeatability of it and, if so, LEV would thus Then be truly possible.

@CANanonymity - at least on a cellular level there doesn't appear to be a point of no return. I remember a few years back, they took cells from people of different ages, including centenarians, and reverted them back to the pluripotent state (iPSCs). They found that there was no discernable difference between someone in their twenties and someone over a hundred. What this means for an entire organ or organism remains to be determined.

By the way, there was yet another podcast with David Sinclair released yesterday, which is longer & better than the one I posted above. After the hour and a half mark, he talks in detail about this and the future possibilities which might emerge - well worth a listen:https://www.richroll.com/podcast/david-sinclair-436/

Hi Chris! Thanks for that..I did some more digging and foudn this...I think this could answer my question, partially and thus gives up much hope.

''Two key insights came when fibroblasts from LAKI progeria mice were analyzed after OSKM expression was terminated. First, it was shown that the age-associated phenotypes return but, strikingly, ****they could be reversed again if OSKM expression was reintroduced****: cyclic expression of OSKM maintained the reversal of age-associated phenotypes''

This would support the repeatability of things and epireprogramming, as something that continuously be repeated/looped forever for it erases (the epimethylation code on DNA) all traces of 'tabs' on aging/by reverting epiclock/epiage.

''...The cyclical regime for OSKM expression was then used in vivo, with expression for 2 days and no expression for 5 days. This cycle could be repeated as often as required. Cyclic expression of OSKM in LAKI mice had a dramatic effect. Not only were age-associated features reversed but there was also a significant increase in both median and ***maximal life span***''

''There was no increase in formation of teratomas or mortality in vivo. "Partial reprogramming" did not lead to the loss of differentiation markers and expression of pluripotency markers such as Nanog, indicating that age reprogramming in vivo can be achieved in the absence of developmental reprogramming''

''On the other hand, doxycycline treatment in LAKI mice not carrying the 4F cassette did not lead to any improvement in the lifespan compared to untreated controls (Figure 4B). Significant improvements in external appearance, including a reduction in spine curvature, were observed in LAKI 4F mice subjected to cyclic OSKM induction (Figures 4C and S4A). Detailed necropsy analyses performed at 14 weeks of age after 6 weeks of cyclic induction of OSKM revealed gross improvement in the appearance of the gastrointestinal tract in doxycycline-treated LAKI 4F mice compared to untreated mice (Figure 4D). Age-associated histological changes are normally observed during physiological aging in multiple organs, including the skin, spleen, kidneys, and stomach (Cesta, 2006; Khanna et al., 1988; Kurban and Bhawan, 1990; Zhou et al., 2008). Importantly, many of these histological changes were improved in 14-week-old LAKI 4F mice subjected to cyclic induction of OSKM during 6 weeks compared to untreated mice (Figure 4E). Doxycycline-treated LAKI 4F mice showed an increased epidermal and dermal thickness and decreased keratinization of the skin compared to untreated controls (Figure 4E). Macroscopic involution of the spleen and lymphoid depletion of the white pulp, visualized by the reduced size of germinal centers, was significantly rescued in LAKI 4F mice after cyclic induction of OSKM (Figures 4E and S4B). Moreover, we observed a decrease in tubular atrophy and interstitial volume in kidneys of doxycycline-treated LAKI 4F mice compared to untreated mice (Figure 4E). Lastly, age-associated loss of parietal cells and thinning of gastric epithelium in the stomach were also rescued by cyclic induction of OSKM''

Clearly, this means morpho-architecture restoring, or least, partial restoring because of the age-related changes seem reversed by OSKM; the ECM is Definitely affected by OSKM (the likes of MMP/matrix metalloproteinase are affected by OSKM), that is a major effect. This would mean that there the morpho dynamic of multiple tissues/organs - Can Be Reversed to that of a younger age (again, at least, partially..it is, Partial Reprogramming, after all, in order to avoid teratoma formation after 11 days of OSKM exposure).

What worries me much more is the fact that OSKM was not enough to make complete abolition of progeria effect in LAKI mouse, so this still succombed of progeria even so - just a bit later instead (like 10 weeks later/a bit less than 3 months later, which in mouse lifespan translates as 10-15% increase of lifespan; in progeric mice at least).

The riddle is what happens in healthy mice, and if OSKM regimen started at birth.

''As was seen with mouse WT 4F TTFs, short-term OSKM induction in human 4F cells significantly reduced the number of foci of histone γ-H2AX per cell compared to untreated cells (Figure 6E). In addition, short-term induction of OSKM in human 4F cells significantly restored the levels of H3K9me3 (Figure 6F).

Collectively, these results suggest that short-term induction of OSKM can ameliorate multiple hallmarks of aging in late-passage WT mouse and human cells, including the accumulation of DNA damage, cellular senescence, and epigenetic dysregulation, thus demonstrating the potential of partial reprogramming for the rejuvenation of phenotypes observed during physiological aging.''

'' Interestingly, the number, intensity, and volume of foci for histone γ-H2AX, a marker of nuclear DNA double-strand breaks associated with aging (López-Otín et al., 2013), were significantly reduced by short-term induction of OSKM in LAKI 4F cells compared to untreated cells (Figures 1B and S1D). Similarly, levels of p53 binding protein 1 (53BP1), which participates in the DNA damage response, were reduced as a consequence of short-term induction of OSKM (Figure S1E). Furthermore, short-term induction of OSKM in LAKI 4F cells downregulated the expression of age-related stress response genes in the p53 tumor suppressor pathway, including p16INK4a, p21CIP1, Atf3, and Gadd45B, as well as the senescence-associated metalloprotease MMP13 and interleukin-6 (Figure 1C). Additionally, senescence-associated β-galactosidase activity was reduced in LAKI 4F cells by short-term induction of OSKM''

At least thagt is good news, and it seems that organ/tissue reversal is possible, even from very aged people. The fact that senescence is Greatly reduced is a demonstration that it affects all type of senescence, including the most important one, Replicative Senescence/Hayflick, possibly in post-mitotic somatic long-lived non-dividing cells. This means, OSKM, is not just partial rejuvenation, it is True rejuvenation, if not 100% (From telomeres elongation, ECM changes reversal, reduction mtROS, reversal of DNA epiclock age, reduction of DNA frags, I would wager reduction of progerin (seen in progeric mice) and also proteasome/lysosome lipofuscin; maybe it is sufficient (even if partial) and repeatable to actually reverse aging in True rejuvenation, no matter if 90 or 60 years old.

Still, until they make the WT mice obtain OSKM and see how much it extends their natural healthy lifespan, the question lingers. Progeria mice do not escape their progeric condition despite OSKM.
My take:
It will depend if OSKM is started, at BIRTH or OLD age of WT mice, and then we will see how much OSKM is really extending longevity in WT mice depending on When it was started in the mouse's lifespan.

1. Age reprogramming and epigenetic rejuvenation
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6300877/

2. Partial reprogramming induces a steady decline in epigenetic age before loss of somatic identity
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6351826/

3. In Vivo Amelioration of Age-Associated Hallmarks by Partial Reprogramming
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5679279/

Imagine the market size if partial reprogramming of people's skin cells made their skin look younger?

Hi #CANanonymity

You always go through an excellent thinking process in your posts

The history of somatic cell reprogramming, going back to the Briggs and King experiments from 1952, highlights the potency of all of these processes in turning back cellular age / identity - https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2946127/

The big mystery is how you deal with with all that was learned in the ensuing "cross-age transplantation" era (1950s-1980s) - which showed that putting "young things" into "old organisms" was not a very good long term strategy towards rejuvenating an organism -

Some good chapters in this book on the history of that work - https://www.sciencedirect.com/book/9780123694393/principles-of-regenerative-biology#book-info

That is the true "black box" of longevity biotech that must be solved

As mentioned above, regenerative organisms wield the simultaneous ability to re-establish the morphodynamic developmental processes, along side embryogenic cellular potential

Re - Thanks for that Ira (and you too do an excellent thinking process), and the links, will check that.
Just a 2 cents. I think we are now at that point, matured enough, that we feel much more confident on the realization/potential and scalability of the rejuvenation from reprogramming; it took a long time (1952 to now (nearly 70 years...but, finally, we arrived). In fact, it's those kind of old papers that put marvel/awe in me - and - also, disbelief at the same time (the more 'less awe/amazed', thus the bit 'depressed' kind of disbelief)...like wow it takes, time, (lotsa) for things to happen (we are patient, but impatience always comes at one point of late, then its depression as how hope fades)...but 70 years is done, and now we are now. So we are eternally grateful to these pioneers of old times for making these small stepping milestones, setting it in place/for it to happen today, some seven decades later. The more I read of science the more it's 'all over the place'...like a miracle Could happen tomorrow...like LEV...let's say...or we could be 'searching for needles in haystacks'...for Another Next 7 decades...that's what it is (who has seven decades?). It also means that we must be 'take it one day at a time' and be optimistic yet realist on things changing in the near-future (near-future, like say, less than 30 years; thus who wants to wait another seven decades). Because of the uncertainty/unassurance/unguarantee of things (biogerontology is a 'odds thing/gamble'/nebulous ambiguous contradictory results 'at odds with self', every day is a gene roulette spin), we must keep realist and hope for best (and prepare for worse too, Should it happen, something we hope not). But, for people that are very sick (right now), it'S even more imperative, they follow this like lazer-beam precision, because time is missing for them.

I think OSKM is our first step in the true biorejuvenation domain; it is spectacular, and I wager, it will only get better as we hone it. LEV then becomes far more a concrete possibility; maybe not LEV, exactly, but if humans obtain 150-300 years extra in life above their max - that will, already, be the most incredible achievement Ever; we are inching closer and closer to curing diseases, aging and death.
I hope to be alive in next few decades to see that happen. Just a 2 cents.

@jimofoz

Hi jimofoz! Just a 2 cents. Indeed, it would be Incredible/Giganormous for sure. When I look at things like antioxidants, supplements, collagen peptides, collagen injection, botox injection, retinoic acid for skin, it is a big market for sure...but I think it will take more than that. We have to convey it's more than just 'skin reversal' of aging..it is Whole Systemic reversal of aging - pushing death as far as possible..that's a lot more meaningful than looking young in the face only. People must understand taht All Organs/cells benefit, thus this affects Entire Aging. This will make them understand the power of this and think ''I can get beautiful young skin..like when I was 20...long ago...but more than that, I can live Long(er), much longer and (maintain) in great health - I feel renewed sense of purpose/desire to live a life, my life, and it will last much longer. I have lots of time ahead now and can do whatever I please for my life will last long''. Kind of like the 'energizer bunny' (battery ad)...it will keep on going..and going..and going....Who wants a 1.5 Volt battery..we all want a 9 Volt - 9 lives Cat Rayovac battery (imagine having '9 lives like a cat').

Just a 2 cats.

My goodness @CANanonymity, I've never seen you this positive before! It's nice to see :-)