AGINGSCIENCES™ – Anti-Aging Firewalls™

An amazing discovery is still in the process of being made.  Exposing the DNA of many kinds of body cells to just four transcription-factor proteins causes a cell to lose all memory of what it is and does and become what is called an Induced Pluripotent Stem Cell (iPS cell), a cell very much like an embryonic stem (ES) cell.  It appears that an iPS cell, like an ES cell, is capable of progressively differentiating into any cell type, be it heart, skin, nerve, bone or muscle.  In other words the four proteins reboot the cell into embryonic stem-cell type pluripotency.  The entire history of epigenomic information that cell has inherited from its progenitors over the entire life of the animal is simply wiped out.  This includes not only information about what the cell has become (e.g. a brain neuron or a bone or liver cell) but also the history of experiences inherited from its progenitors going back to the inception of the animal.  This history, expressed via methylated DNA can strongly condition the behavior of the cell.  See the Feb 28 post in this blog on Epigenetics, Epigenomics and Aging.  

The proteins are Oct4, Sox2, Klf4, and c-Myc.  Oct4 and Sox2 are transcription factors known to have to do with ES self-renewal and proliferation.  Klf4, and c-Myc are transcription factors known for their roles in cell proliferation, differentiation and survival and regulating gene expression.  This reprogramming effect was at first reported in 2006 and limited to starting with mouse fibroblast cells carrying engineered selection markers(ref,ref).  Later, the effect was demonstrated using a retrovirus to transport the proteins into the nuclii of mouse cells(ref) without a need for engineered cells.  However, the retrovirus could also inject its own DNA into the chromosomes affected, so the process appeared to be unreliable because it could affect the genome of the cell.  More recently, it has been possible to induce pluripotent stem (iPS) cells from mouse fibroblasts and liver cells by using relatively benign nonintegrating adenoviruses(ref).  It appears in several experiments that the four proteins wipe clean all the lifelong DNA epigenetic modifications that normally occur as a stem cell progressively differentiates into being a very specific kind of body cell, like a hair follicle cell.  “DNA methylation, gene expression and chromatin state of such induced reprogrammed stem cells are similar to those of ES cells.”

This discovery is still in process because most experimental work so far has been with mouse cells and it is still not known for sure how completely  similar the behavior of iPS and ES are.  Nobody has been able so far to make a mouse out of an iPS cell, for example. The possible implications for regenerative medicine are immense.  The idea is to remove some cells from a person, say a small tube of blood cells, restore these in the lab to being iPS cells and then use these in lieu of ES cells for organ regeneration in that person.  Because the cells are from the same person, there would be no problem of immune system rejection.  Several issues require resolution before this kind of therapy can be practical, however.  First, the pluripotency and safety of using human iPS cells must be confirmed.  There is a possibility that they could contain genetic remnants of the andovirus vector that could create mischief for example.  Second, it is important to find effective ways for introducing the iPS cells into human organs so they achieve the desired results of organ regeneration.  If ES or iPS are randomly injected into body tissues they are likely to form teratomas, ugly encapsulated structures of varied body tissues including lung, heart, liver or brain tissue, or bone, teeth or hair.  Careful attention has to be paid to signaling structure to assure the ES or iPS make the kind of tissues desired and only that kind.  This is a fundamental issue of organ regeneration via stem cell therapy whether ES or iPS cells are used.  Also I am unclear about another important matter.  What happens to the telomeres in a cell from a mature person when it is converted to an iPS cell?  Do the telomeres stay short reflecting the age of the donor or are they somehow enlongated as they would be in an ES cell?  Does an iPS cell have sufficient expression of telomerase when it differentiates to carry it through the large number of generations required for effective organ renewal?  As far as I know, these are issues still to be researched.

In terms of aging, the four proteins seem to turn the clock back in a cell to ground zero.  That is, all the epigenetic information (DNA methylation) that cell and its parents, grandparents, great-grandparents, etc.  gathered in the process of many rounds of cell differentiation and division and life experience is completely wiped away in the process of it becoming reprogrammed as an iPS cell.  For halting or even reversing human aging we don’t want that to happen to most of the cells in our body; it would turn us into blobs of undifferentiated stem cells.  What we might like, however, is selectively to erase epigenetic information related to programmed aging while retaining epigenetic information related to cell differentiation.  Or, more simply put, we would like selectively to reset our cells to a younger state while keeping the degree of cell differentiation appropriate to that younger state.  See the discussion of the Programmed genetic changes theory of aging in my Anti-Aging Firewalls treatise.  This is a challenge of a quite different order of magnitude than is simple rebooting. 

We can probably expect important new research results related to iPS cells in the coming year, months or even weeks. 

A reported large-scale population study by Scottish researchers indicates that longevity is highly correlated with childhood intelligence quotient, especially for people who grow up in poorer neighborhoods.  A thousand people were followed during a 70-year span.  During a 25 year interval  “— 51 percent of the men and 38 percent of the women in the study died. In simple terms, there was a 17 percent greater chance of death for every 15 points of lower childhood IQ. After adjusting for deprivation and social class, this difference was reduced to 12 percent.  These adjustments separated socioeconomic effects from IQ and explained some, but not all, of the differences associated with lower IQ.”  The reasons for this effect are not clear.  One possibility according to the study authors is that poorer people with lower childhood IQ lead more deprived lives and are more vulnerable to diseases and other causes of death.  Another possibility is that low childhood IQ combined with poverty was correlated with low childhood health in the first place leading to increased mortality.  I suggest a third possibility: that members of the higher IQ group had more willingness, commitment and capability to pay attention to their health and longevity and that this effect transcended socioeconomic class.

The nuclear transcription factor NF-kappaB plays a prominent role in one of the advanced theories of aging, Programmed genetic changes.  Several new pieces of research highlight the mechanisms by means of which this multi-faceted substance impacts on aging and the importance of the anti-aging firewall substances that inhibit the expression of NF-kappaB.  I mention two such studies.

One study has to do with the role of Tumor Necrosis Factor (TNF) activating the expression of NF-kappaB in Muscle Progenitor Cells (MPC).  This study is also relevant to the fourteenth theory of aging, Decline in adult stem cell differentiation.  TNF acts via a number of pathways in a complicated manner including activation of NF-kappaB to produce a variety of effects including induction of apoptosis (cell suicide).  This apoptosis is useful when cancers are concerned but is also potentially destructive of healthy cells. It appears that in MPC cells at least, TNF-alpha activates NF-κappaB more in older animals than in younger ones.  This leads to apoptotic signaling and death of MPC.  The problem is thought to be a decline in age of effective cellular mechanisms for keeping NF-kappaB inactive.  Practically speaking, this appears to support the importance of the thirty-nine anti-aging firewall substances that inhibit the expression of NF-kappaB.

A second study has to do with the role of the sirtuin SIRT6 in regulating the expression of NF-kappaB.  Another member of the sirtuin family, SIRT1, has been extensively studied and even sometimes referred to as a key “longevity gene.” But SIRT6 also seems to play an important role in keeping NF-kappaB expression in its proper place.  SIRT6 works with NFkappa-B to control the activity of genes connected with metabolism, inflammation, immunity and aging. When SIRT6 is in short supply, NFkappa-B becomes hyperactive and turns up activity of aging-linked genes.  “We propose that SIRT6 attenuates NF-kappaB signaling via H3K9 deacetylation at chromatin, and hyperactive NF-kappaB signaling may contribute to premature and normal aging.”  Again, taking the firewall substances that control expression of NFkappa-B may be an effective anti-aging measure.

The third theory of aging covered in my Anti-Aging Firewalls treatise is Mitochondrial DNA Mutation.  Research reported today relates to the relationship of mitochondrial dysfunction to Parkinson’s Disease (PD).  Other new research indicates that taking two substances in the anti-aging firewalls that act on the mitochondria may prove to be a safe and effective strategy for preventing PD. The new research indicates that an inherited form of PD is caused by mutations in the PINK1 gene which is localized to mitochondria. The researchers found that insufficient expression of PINK1 leads to an aberrant calcium overload inside the mitochondria.  The calcium overload in turn results in the production of Reactive Oxygen Species (ROS) that interfere with the ability of the mitochondria to transport sugar needed for energy production.  The result can be injury to and death of dopamine-producing neurons leading to Parkinson’s Disease. 

Since excess mitochondrial ROS are involved, it seems plausible that antioxidants that act in the mitochondria might help prevent PD. We have known for a few years from the pioneering work of  Bruce Ames that a combination of alpha-lipoic acid and actyl l-carnitine and alpha-lipoic acid act together powerfully in such a capacity.  Many studies have confirmed the power of these two anti-aging firewall substances for brain health when taken together.  “Dietary supplementation of young and aged animals increased the proliferation of intact mitochondria and reduced the density of mitochondria associated with vacuoles and lipofuscin.  Feeding old rats ALCAR and LA significantly reduced the number of severely damaged mitochondria (P = 0.02) and increased the number of intact mitochondria (P < 0.001) in the hippocampus.  These results suggest that feeding ALCAR with LA may ameliorate age-associated mitochondrial ultrastructural decay and are consistent with previous studies showing improved brain function. (ref)”  A research report originated in a Chinese university looks more directly at how these two substances might prevent PD:  Combined R-α–lipoic acid and acetyl-L-carnitine exerts efficient preventative effects in a cellular model of Parkinson’s disease.

Back in 1995 my friends mostly humored me when I told them I was planning to live 165 more years and the secret to my success would be connected with future research that would allow me to extend my telomeres.  “Tele-what?” they said.  “Does that have to do with communications?” This was 10 years after the discovery of telomerase in 1985 by Elizabeth H. Blackburn and Carol W. Greider.   In 1995, the roles of telomeres and telomerase in cell biology was sometimes thought by scientists to be interesting but mostly thought to be of rather peripheral interest.  That is, except for a few visionaries like Michael Fossel who grasped the importance of these topics early-on. Now we know that telomeres and telomerase are of central relevance with respect to cancers, stem cell differentiation and longevity.  And of course Telomere shortening is one of the most important theories of aging covered in my Anti-Aging Firewalls treatise.  

Today, Rockefeller University announced the winners of the fifth annual Pearl Meister Greengard Prize to Blackburn, Greider and Vicki Lundblad of the Salk Institute for Biological Studies.  The prize is being awarded for the discovery of telomerase and for studies of its regulation.  In 2008 Blackburn and Greider received the Paul Erlich and Ludwig Darmstaedter Prize, the leading science prize in Germany, for the same work.  Greider won the 2007 Dickson Prize in Medicine for her contribution. Blackburn and Greider also received the Wiley Prize in Biomedical Sciences in 2006.  The two shared the Louisa Gross Horwitz Prize 1n 2007, and with Jack W. Szostak received the Lasker Prize in 2006.  The honors and prizes are likely to keep rolling in.  It is interesting that it has required more than 20 years for the seminal work of Blackburn and Greider to receive the acknowledgement it deserves.  Some of the other advanced longevity research going on today may likewise not be fully acknowledged until 20 or more years from now.

The suggestions in my Anti-Aging Firewalls treatise, for following certain lifestyle patterns and taking certain supplements for longevity, are based on scientific research rather than simply on folk remedy lore or conventional wisdom.  But what is the nature of this scientific research on which I base my suggestions?  It can be of several different kinds, including:

1.     Macroscopic studies of large populations.  These studies involve looking for correlations among selected factors.  An example is a study of centenarians on Okinawa, an island where an unusual number of people live a long time.  This study of some 900 centenarians, their families and control groups helped identify specific genes and gene polymorphisms that appear to contribute to longevity as well as contributing lifestyle factors, including eating relatively few calories, exercising and not smoking or consuming alcohol.  And of course in Okinawa people eat lots of fish and drink green tea.  These studies can reveal interesting correlations and clues.  For example, the Okinawa study established that the centenarians studied have genetic polymorphisms that place them at lower risk for inflammatory and autoimmune diseases. 

2.     Large population cohort studies.  These are studies that follow coherts of thousands, tens of thousands or even hundreds of thousands of people over 10-20 or longer year periods, like the Woman’s Health Initiative or the Framingham Heart Study or the Bogalusa Heart Study.  Again, they look for correlations such as the effect of smoking or being overweight on cancer incidence or longevity. These studies have been particularly useful for establishing the validity of conventional wisdom as related to longevity, such as clearly documenting the effect of having a positive mental attitude on longevity.

3.     Controlled clinical trials.  These are carefully controlled double-blind studies typically pursued for drug certification that proceed in phases.  They may typically  involve anywhere from a few dozen to thousands of carefully selected people over a test periods of several months for the final phase.  They are usually very narrowly focused and yield limited information with respect to longevity.  For example, a list of clinical trials for patients diagnosed with gliablastoma, an incurable brain disease, can be found here.  These trials are very specific with respect to substance being tested, patient conditions and their relationship to other therapies.  Clinical trials typically cost tens or hundreds of millions of dollars so there is no incentive for a drug company to study a promising natural substance that is in the public domain. Also, for longevity purposes, a six-month study is not likely to tell much.  If we wanted to test some kind of longevity concoction that we thought would double human lifespans, a clinical trial would have to be run for 40-100 years to yield meaningful results.   

4.     Animal experiments.  Mice and rats are genetically very similar to humans but live only 2-3 years, so are excellent subjects for longevity-related experiments.  The studies can be quite technical.  Here is an example relevant to the cell nuclear factor NF-kappaB known to be relevant to human longevity: Maintenance of NF-κB Activation in T-Lymphocytes and a Naive T-Cell Population in Autoimmune-Prone (NZB/NZW)F1 Mice by Feeding a Food-Restricted Diet Enriched with n-3 Fatty Acids We know of several approaches that can extend the normal lifespan of mice by 30% to 50%  We are not sure how many of these approaches will scale-up to work for humans but these experiments are providing valuable clues and are a source of optimism for longevity aficionados like me.

5.     In-vitro and in-vivo studies of cell populations.  There is a great numer of experimental studies going on that look at specific cell populations under particular conditions that that have a bearing on longevity.  For example, many such studies look at neurogenesis and adult stem cell differentiation as impacted by specific gene activation cascades and particular proteins, or as stimulated by certain dietary substances.  These studies can yield specific nuggets of insight such as the roles of key proteins and activation factors like INK4a, P-53, and NF-kappaB. Again, there are very many of these studies and they can be quite technical each yielding a single piece of the immense longevity puzzle.  An example related to eating pigmented fruits is A dietary anthocyanidin delphinidin induces apoptosis of human prostate cancer PC3 cells in vitro and in vivo: involvement of nuclear factor-kappaB signaling.

6.    Synthesis and review studies.  These are studies that consider results together from possibly many experiments and look at them in terms of the powerful forefront areas of genetics, cell signaling cascades, gene activation, genomics and epigenomics.  They also draw on knowledge from related areas, such as computational genomics, epigenomic and protein-folding databases.   Some of these studies are starting to link gene expression factors to longevity, such as in SIRT6 Links Histone H3 Lysine 9 Deacetylation to NF- B-Dependent Gene Expression and Organismal Life Span.  We have identified various signaling cascades directly related to longevity such as the Insulin Growth Factor 1 axis.  This axis seems to be the one that is involved in achieving longevity via calorie restriction.  And it also seems to be activated by taking the supplement resveratrol. 

All the actions and supplements in the anti-aging firewalls are based on one or more of these kinds of research. Most are supported by several of these kinds of research and, for a few firewall elements, supporting research exists on all of the above levels.  For example, the value of green tea as a cancer preventative is established on all of the above levels as is the value of regular hard exercise for longevity in general. 

It is clear that radical life extension—to beyond age 110—must depend on knowledge associated with the newer and more sophisticated ongoing studies in epigenetics, molecular biology, and medical research.  Research suggests that certain substances already in the anti-aging regimen may act powerfully toward this end, but what they can actually do for human life extension will not be clearly known for many years.  These substances include:

–         Use of r-alpha lipoic acid and acytl-l-carnitine to address cell mitochondrial longevity and inhibit unwanted cell apoptosis (self-destruction).

–         Use of resveratrol or resveratrol homologs to activate the SIRT1 and FOXO3 “longevity” genetic pathway, the pathway known to confer life extension due to calorie restriction

.–         Use of astragaloside IV or cycloastragenol  to activate telomerase expression in body cells, possibly immortalizing these cells and conferring longevity to the associated organs.

.–         Use of combinations of green tea, curcumin, and other phyto-substances for their powerful cancer-preventative effects and cardiovascular benefits that operate through genetic mechanisms.

Longevity research is a ride into uncharted territory.  The ride is moving faster and faster and I believe we will see breaking the 120-year ultimate human age limit in my lifetime.  As a matter of fact, I am planning to be one of the breakers.  

There were a number of press reports this morning on a finding based on the Woman’s Health Initiative data, a study of over 100,000 woman that started in 1994.  The study shows that a piece of conventional wisdom often found in touchie-feely books is in fact correct: positive mental attitude enhances longevity; negative attitude reduces it.  Pessimistic woman, those with a dim outlook on life, were 30 percent more likely to die from heart disease and 14 percent more likely to die from any cause than optimistic ones.  Cynically hostile women, ones who tend to mistrust people, were 23 percent more likely to die from cancer and 16 percent more likely to die from any cause.   Optimistic women were also less likely to smoke cigarettes or have high blood pressure or, diabetes.  Yet again, the message is that your mental state can create epigenomic modifications, DNA methylation on your chromosomes and histone acetylation/deacetylation modifications, and therefore alter your gene expression pattern and therefore affect your longevity.  There is no more mystery to it.

The first P is Perspective.  To start, you need to have a perspective that a very long and healthy life is possible, that you want it and that you are willing to take whatever actions as are necessary to have it. 

The second P is Participation.  Participate actively in managing your health and creating your longevity.  This involves being knowledgeable about your own conditions and the options available to you and seeking out and taking advantage of resources needed for your wellbeing and longevity.  These could include securing good foods, companionship, intellectual challenges, exercise, dietary supplements and knowledgeable medical assistance, to start the list. 

The third P is Proactivity.  Simply put, go out there and create your own health and longevity.  And keep creating it. It won’t happen automatically. 

The fourth P is Prevention.  There are multiple ways to avoid diseases and slow or stop aging processes outlined in my Anti-Aging Firewalls treatise.  Take advantage of them while you are healthy.  Once you have a cancer, a serious cardiovascular problem, diabetes or senile dementia, your options will be much narrower and not necessarily very good ones. 

The fifth P is Perplexity.  Yes, perplexity!  The processes required for health and longevity are very complex and many are still poorly understood.  You have to do a lot of things to keep healthy and make it to a very old age.   The roadmaps are relatively poor and frequently change.  There are many options and blind alleys, phony nostrums and cures that can take you off course.  Be prepared to be perplexed and confused along the way.  If you take on your longevity as a serious problem to be solved and grapple with it, this will just by itself enhance your neurogenesis and help you keep going longer.  (See the previous blog entry Tough learning and neuron survivability). 

The sixth P is Personalization.  Your health and possible longevity is a highly personalized matter, different than anyone else’s.  This means you need to get to know and monitor your own body’s health parameters.  To start, get regular lipid and and C-reactive protein profiles.  Monitor your heart rate and blood pressure if appropriate.  If you have a blood glucose level issue, you can monitor that daily too.   And take appropriate actions.   If your CRP is off-scale and indicative of inflammation you can increase your use of anti-inflammatory supplements, for example.  And pay special attention for incipient disease conditions like mild arthritis, trigger finger or carpal tunnel syndrome.  These too indicate inflammatory conditions.  If you are getting frequent colds or suffering from minor infections, these could indicate a weakened immune condition.  You might need more rest or to take immune system strengthening supplements.  If you are gaining weight or feeling lethargic you might want to exercise more, etc. And as the availability of gene-chip profiling for susceptibility to disease conditions increases, be prepared to take advantage of this technology for knowing yourself too. 

The seventh P is perserverence.  You need to persevere with your good eating habits, avoiding junk or bad food, non-smoking, daily exercise and taking your supplements, paying attention to all the factors impacting on your health and lonngevity.  Otherwise you might not live long or healthily enough to take advantage of new longevity breakthroughs from molecular biology or nanomedicine as they come along.

In a previous post Protein origami and aging I mentioned how proteins fold themselves up in complex shapes as soon as they are formed and how stress often leads to the misfolding of proteins, a process that can accelerate with age creating vulnerability to a number of dysfunctional and disease conditions.  Misfolded proteins are generally bad news.  In fact, I asserted that if I were to add a 15th theory of aging to my Anti-Aging Firewalls treatise it would possibly be Misfoldings of proteins.  I have posted a long and rather technical note on the protein folding subject in that treatise.  Some proteins have very complex topologies involving knots and slipknots. It turns out that the processes of folding can involve intermediate configurations with slipknot transition states and the folding trajectory can even involve backtracking.  The presence of these can affect the probability of successful folding.  Modeling of the folding process for complex proteins is starting to yield insight of how it can work. 

The March 2009 issue of Scientific American reports research on what happens to neurons after neurogenesis in rats. Under normal circumstances thousands of new neurons are generated every day in the dendrate gyrus of the hippocampus. Within a few weeks most of those neurons die if the animal’s life is unremarkable. However, if the animal is confronted with a sufficiently daunting and important learning task and successfully learns something complex and new, then many of the neurons will stay alive. This appears to be particularly applicable to learning that affects future behavior. The “use it or lose it” concept for maintaining cognitive capability seems to relate directly to the survivability of neurons.  See the neurogenesis discussion in my Anti-Aging Firewalls paper.