The Life Extension Pathway, Resveratrol etc.

and Cancer
Control: Mitochondrial Biogenesis Duality, the Metabolic
Mechanism and Practical Applications.
Gregory S. Bambeck Ph.D. and Michael Wolfson J.D., M.B.A.
Kent, Ohio U.S.A. 44240

SUMMARY STATEMENT: Cancer, heart disease and diabetes II, the

three largest killers of the first and second world nation’s human beings
(85%), and their disease antithesis, a healthy squirt from the fountain of
youth, are finally defined under a singular unifying global hypothesis.
Experimental molecular mapping proves that the regulatory pathway
mechanisms define it as a true, real and clinically demonstrated system for
the first time. Publicly available, practical and easily workable disease
blocking and life extension implementation are readily available to anyone.
There are three sections: ABSTRACT; PATHWAY MECHANISMS;
PRACTICAL APPLICATIONS. Read on!

ABSTRACT

Full mitochondrial biogenesis is a two phase temporal process consisting of

an early phase primarily associated with anabolism, cell replication and the
making of over a thousand constitutive mitochondrial proteins that create
new, but NADH to OX/PHOS inefficient mitochondria. Between these
replication events, cell homeostatic controls up regulate a late phase set of
mitochondrial respiratory chain proteins that create efficient NADH to
OX/PHOS associated with a shift toward catabolism and autophagy of
dysfunctional mitochondria and other cell debris. The early phase
(neogenesis) supports cell growth, rejuvenation and whole body vitality in
the short term, while the late phase (regenesis) supports cellular
housekeeping and repair functions in the life extending long term. The
immediate upstream effector of mitochondrial biogenesis is the
mitochondrial proliferator co-activator (PGC-1alpha). Its up regulation
institutes neogenesis and its down regulation institutes regenesis. Caloric
restriction (CR) activates regenesis by up regulating adenosine
monophosphate activated kinase (AMPK), while cancer activates
neogenesis in the absence of regenesis by down regulation of the same
AMPK pathway, upstream of PGC-1alpha. Recent ‘rediscoveries’ show
that a cancer cell metabolism proposal of 1980, is correct in its many
metabolic particulars. Most cancer cells are mutationally glycolytic fetal
enzyme driven ‘sugar junkies’ supported by obligate mitochondrial ATP
production inefficiency. Cancer cells are stuck in this cell growth drive
state (metabotype), and become relentlessly replicative under the influence
of mitogens. Recent genuine discoveries show that inhibition of the life
extending CR pathway supports this ‘sugar junkie’ growth state by creating
and maintaining inefficient and neogenic mitochondria in the presence of
forced hyperglycolysis. Blocking fetal glycolysis and re-establishing the
CR pathway pattern creates the regenesis of efficient mitochondria and
halts cancer cell growth, and can sometimes even initiate cancer cell
apoptosis. We describe the pathway mechanism as a unidirectional
feedback loop starting with the CR target, AMPK, and how it regulates
mitochondrial biogenesis, the reactive oxygen species (ROS) output, of
which, feeds back to AMPK. We also make sense of CR mimetic
resveratrol, and its bioavailability in this context, and further employ these
understandings to envision simple nutriceutical and lifestyle synergies to
fight cancer, the major diseases of aging and even aging itself.

PATHWAY MECHANISMS

To make this easy to understand, we wish to start with some rules of the
road. First, when we use the adjective ‘chronic’ or prefixes like ‘hypo’ or
‘hyper’, we are referring to aberrant, unnatural or man-made over impacts
upon normal homeostatic systems or typical physiological shiftings in
standard metabolic systems. These words are used to emphasize a principal
or powerful effect of some kind. Second, since CR is the ‘gold standard’ of
life extension, we will use it as a ‘home base’, or reference point, that most
of our forays will diverge from, and then return to. Third, we will focus
primarily on the unidirectional multi-toggle switch feedback cycle from
AMPK to target of rapamycin (TOR) to PGC-1alpha to ROS to sestrin
(SESN) and back to AMPK, with the up regulation of AMPK effectively
down regulating every other component of the cycle downstream of it.
Since the system is a closed feedback loop, everything is upstream of
everything else as well as downstream of everything else, like the
proverbial snake that eats its tail. However, extrinsic factors affecting any
component can over ride their immediate upstream regulators, as we will
often point out. Memorizing, or keeping a note pad with this simple
AMPK, TOR, PGC-1alpha, ROS, SESN circuit, as a principal reference
point, keeps the discussion grounded.

We will begin with a brief outline of the CR pathway. Most simply put, CR
activates AMPK which down regulates TOR. Down regulated TOR shifts
cell metabolism away from anabolism and toward catabolism, initiates
autophagic clean up of cell debris, such as dysfunctional mitochondria and
down regulates PGC-1alpha. When down regulated, PGC-1alpha results in
the regenesis of existing mitochondria by initiating the transcription of
mitochondrial respiratory chain proteins that efficiently link NADH to
OX/PHOS ATP production. This causes mitochondrial ROS production to
fall, which in turn, down regulates the ROS sensor, SESN. SESN then
down regulates its stimulation of AMPK, causing the circuit to rebalance
back toward its homeostatic center. Note here that active SESN up
regulates AMPK and inactive SESN fails to activate, so its up regulation is
active while its down regulation is passive. Thus, continued CR will bypass
SESN and chronically up regulate AMPK to constantly enhance the CR
pathway to reduce mitochondrial ROS. Elevated ROS is the principal life
shortening component in the system, and thus, its repression is the largest
single life extender known. This example demonstrates that increased
AMPK activity causes a decrease in all the other four (TOR, PGC-1alpha,
ROS, SESN) components in the unidirectional feedback loop. Conversely,
decreased AMPK activity, as in p53 dysfunctional cancer cells, causes the
up regulation of the same four components down stream of it, which
incidentally, causes the neogenesis of inefficient mitochondria.

As exemplified by our CR model, AMPK to TOR to PGC-1alpha are the

principal mitochondrial biogenesis inputs and their responses, while ROS
to SESN are the principal mitochondrial outputs and their responses. We
know that there are many steps between each of these five major toggle
switches and that there are also myriads of branching pathways and gene up
and down regulations stemming into and from each toggle switch, but they
will be mentioned in passing only as needed, because it is the central
unidirectional cycle that is critical to the cancer, diseases of aging decline
and life extension metabolics we key on, herein. Very elegant and
evolutionarily apt extensions of this central and related circuits and their
nutrient sensing pathways can be found in Science, vol.327, 3/5/2010
p.1210 and vol.328, 3/16/2010 p.324. Lastly, the central focus, here, is on
cellular bioenergetics and metabolics because these functions hail back to
single cell eukaryotes and have finally earned their seat at the cancer cell
control systems round table along with telomeres, growth factors, apoptosis
etc. Later, we will see how the AMPK, TOR, PGC-1alpha, ROS, SESN
circuit is the exact same circuit controlling cancer and life extension, albeit
operating in the opposite direction. We shall also see that cancer ‘cure’, or
at least control, and CR share the exact same circuit when operating in the
same direction, with mutationally driven hyperglycolysis being the lone but
critical cancer stand out. But first, let’s look at each major component in
the system.

AMPK stands at the headwaters of the CR pathway. AMPK monitors

cellular energy charge by being sensitive to the AMP/ATP ratio, which
generally represents the fuel nutrient availability in the cell. High
concentrations of the energy rich ATP molecule represent fuel nutrient
sufficiency and concomitant low AMP. Conversely, high AMP and low
ATP indicate low fuel availability as in CR, which up regulates AMPK via
AMPK kinase. Increases in AMPK can inhibit TOR by countermanding the
growth factor pathways that activate TOR. Things that activate AMPK
inhibit TOR, and things that inhibit AMPK activate TOR. Things that
activate AMPK initiate the CR pathway and support life extension and
inhibit the cancer metabotype. Standing directly upstream of AMPK, SESN
activates AMPK when it self is activated by the cancer cell growth
suppressor (p53) or by ROS, mostly of mitochondrial origin. The P53
protein is pro-apoptotic and its inhibition or dysfunction is found in about
half of all cancers. Components or systems that reduce AMPK activity
shorten life expectancy and promote the cancer metabotype. CR mimetics
extend life expectancy and inhibit the cancer metabotype.

This last effect is exemplified by the anti-type II diabetic drug, metformin

which is a direct activator of AMPK, causing the typical CR reduction in
insulin resistance and the downstream inhibition of TOR. Metformin has
been found to increase life expectancy in cancer victims, diabetics and
healthy animals. Direct activation of AMPK by metformin enhances the
ROS activated SESN switch or can over ride SESN inactivation by faulty
p53. Intracellular concentrations of resveratrol in the 20 to 50 uM range
activate AMPK similarly to metformin. Dietary free resveratrol rarely
attains concentrations above the low single digits of uM. Metformin is our
best known chemical example of a CR mimetic.

Now, let us take a brief tour of TOR. In our unidirectional metabolic loop
activator model, TOR is the first primary target downstream of AMPK.
TOR is a major NADH/NAD redox sensor and a master metabolic
regulator switch involved in a dizzying array of integrated pathways, which
can be found in a web search under ‘mammalian target of rapamycin’.
Fortunately, several major converging and diverging pathways impinge
upon and emerge from TOR, allowing us to simplify matters. For instance,
mitogens, growth factors, hormones and AMPK act upon TOR through a
common intermediate called TSC2. Thus, in a functionally simplified
sense, TOR responds to a delicate balance of AMP/ATP energy charge,
NADH/NAD redox state, fuel nutrient availability, mitogens, growth
factors, growth factor suppressors and genotoxic ROS load. TOR outputs
are as multifunctional as its inputs, but are most commonly associated with
the integrated regulation of mutually exclusive anabolic or catabolic
systems.

The down regulation of TOR by elevated AMPK activity (by CR, for
example) institutes a shift toward catabolic efficiency in support of the
caloric restriction demands for maximum energy output from maximum
fuel conservation under nutrient fuel limiting conditions. In this regimen,
another TOR pathway institutes increased mitochondrial respiratory
efficiency through the regenesis pathway, and still another TOR pathway
up regulates the autophagy of cellular debris and dysfunctional
mitochondria. Cellular efficiency, ROS reduction and elevated
housekeeping functions increase life expectancy and foster disruption of
the cancer metabotype. This process can also be enforced by the TOR
inhibitor and foreign tissue rejection suppressor, rapamycin. Rapamycin
can facilitate life extension by pulling an end around AMPK and directly
inhibiting TOR to switch the cellular drive state from anabolism to
catabolism, and also very importantly, push mitochondria into the state of
regenic efficiency by down regulating PGC-1alpha.

Finally, as the last major element in the mitochondrial biogenesis input side
of the unidirectional loop, a short summary of PGC-1alpha function might
be in order. When PGC-1alpha is activated, it turns on over a thousand
genes which are devoted to the very complex but singular mega-function of
building more mitochondria (neogenesis). As we saw, when TOR is up
regulated, it in turn, up regulates PGC-1alpha, and the building of all of the
mitochondria except the respiratory chain, is instituted. The respiratory
chain transcription elements can only be constructed when PGC-1alpha is
down regulated, later. Chronic stimulation of PGC-1alpha via chronic up
regulation of TOR by growth hormone keeps mitochondria in a state of
neogenesis with impoverished regenesis, which is a high ROS generator.

This nowhere more manifest than in the cancer cell metabtype. When
mitochondrial respiratory NADH to OX/PHOS coupling is compromised
by neogenesis without regenesis, the mitochondria, although primarily
catabolic in function, actually enhance anabolism by supporting increased
glycolisis and, thereby, shift the balance of glycolytic products,
mitochondrial feedstocks and NADH reducing power toward cell building,
all supported by an increase in the glycolytic to mitochondrial ATP
production ratio. In his dissertation at Kent State University on
mitochondrial alterations in a lymphoblastic lymphoma transplanted into
DBA/1J mice, in 1980, Bambeck reviewed dozens of cancer type cell
mitochondria, and saw a pattern. He provided a detailed catabolic chart, a
general NADH cell nutrient and building block flow chart and an extensive
narrative describing these connections. The AMPK, TOR, PG C-1alpha
mitochondrial input side of the CR pathway, only now available, finally
supply the control elements that prove his hypothesis, which we have just
begun to re-discover in the last two years. It is surprising how accurate this
thirty year old description is, considering there was no awareness of
AMPK, TOR, PGC-1alpha or the vast array of intermediate connections in
this regulatory pathway, at that time. However, even today, our new
knowledge still boils down to the control of redox, energy charge and
metabolites, which could be monitored even, at that time, in the absence of
this modern regulatory pathway knowledge. This was done by constructing
metabolic flow charts under different drive states and deducing where
control links must exist, even if they were, as yet, unknown.

Just as metformin can bypass SESN by directly up regulating AMPK and

rapamycin can bypass AMPK by directly down regulating TOR, T3
thyroxine in its non-shivering thermogenesis mode, can mostly bypass
TOR by up regulating PGC-1alpha. In addition to ignoring the TOR
anabolic/catabolic toggle, this can help illustrate the temporal duality of
PGC-1alpha function.

A single dose of T3 thyroxine binds to nuclear DNA upstream of the

mitochondrial biogenesis activator master control system and the PGC-
1alpha binding co-activator, and begins transcription. Within five hours,
activated PGC-1alpha causes more than 1,000 nuclear genes that are
specific to the building of new respiration impoverished mitochondria to
begin transcription. These very long mRNAs contain the code for their
respective gene products as well as the leader sequences for their respective
mitochondrial targets. These mRNAs are exported from the nucleus to the
cytoplasm to form translational polysomes where mitochondrial outer
membranes become confluent with endoplasmic reticulum. Depending on
their signal sequences, these mitochondrial proteins are ferried to their
mitochondrial home compartments to take up functional residence. After
about 48 hours the thyroxine and PGC-1alpha signal system decays and a
late set of nuclear and mitochondrial genes specifying the components of
the respiratory chain are activated, causing these poorly coupled neogenic
mitochondria to become efficient ATP producing regenic mitochondria. In
chronic hyperthyroidism, the neogenic phase is powerfully up regulated
relative to the regenic phase and the resultant ROS damage is severe
enough to dramatically shorten life. A common term for early
hyperthyroidism termination is ‘burnout’, because unlike TOR and its
anabolic activated neogenesis function, the thyroxine activation is
catabolic.

In the thyroxine activated neogenic phase, there are also a set of uncoupling
proteins (UCP) that are produced that allow NADH protons to ‘leakback’
into the mitochondrial matrix without their chemiosmotic potential being
captured and stored as high chemical energy ATP. Although this an over
simplified description, the net result is that the uncaptured energy becomes
manifest as heat, which is easy to measure. Thyroxine also supports
catabolism by shifting fuel sources from low energy per carbon sugar to
high energy per carbon lipid, and if need be, protein. However, when PGC-
1alpha is up regulated by TOR, anabolism is supported and glucose is the
preferred fuel. This difference becomes not only important, but highly
magnified when we consider the cancer metabotype, later. Chronic up
regulation of PGC-1alpha, whether anabolically driven by growth hormone
stimulation of TOR or catabolically driven by direct stimulation of
thyroxine as in hyperthyroidism, results in an incomplete mitochondrial
biogenesis stuck in a high ROS producing neogenic state which can induce
cancer and shorten life span via ROS produced genotoxicity and stochastic
randomization of proteins compounded by the absence of phagocytic
housekeeping functions and DNA repair systems. Chronic neogenesis in
the absence of regenesis is not only a cancer cell metabotype inducer but a
cancer cell metabotype maintainer, as well.

Another way to uncouple NADH from ATP production is for mitochondria

to export NADH to the cytoplasm via the NADH/NAD shuttle system. In
growing and dividing cells, the redox and energy charge states always lag,
so this system is employed more heavily. Also, in non dividing quiescent
cells, citrate in excess of krebs cycle needs can inhibit the ‘committing’
enzyme phospho fructo kinase (PFK) at the headwaters of glycolysis. Fetal
PFK is often elevated in cancer cells. Although it is an inefficient ATP and
NADH producer, glycolysis can run at explosively fast rates when
stimulated. For the sake of brevity, we will not distinguish between the two
uncoupling forms in this document.

The above described AMPK to TOR to PGC-1alpha portion of the closed

unidirectional loop are the inputs to the mitochondrial neogenesis/regenesis
system. The ROS to SESN portion represent the ROS output feedback loop
to AMPK. Although the exact mechanism of the mitochondrial output of
ROS to SESN is still to be articulated, its requisite path and net results are
unambiguous. Increases in ROS eventually leads to activation of SESN,
which in turn, activate AMPK to inhibit TOR to down regulate PGC-
1alpha to institute mitochondrial regenesis to reduce ROS. SESN gene
knockout drosophila die prematurely of age related disorders, and the anti-
oxidant vitamin E protects life length function in SESN gene knockouts.

The system, as so far described is more elucidative of life extension than it

is of the generation and maintenance of the cancer metabotype, soon to be
discussed. Basically, life extension via the CR upregulation of AMPK
occurs, mostly due to the reduction of ROS caused by mitochondrial
regenesis of efficient ATP producing mitochondria. This essentially puts
the adult organism in an extended holding pattern until nutrient energy
supplies can support fecundity and other energy intensive functions such as
immune surveillance, wound healing and muscle building.
Here is how resveratrol probably fits into the picture. In dietary quantities,
free resveratrol enters interstitial cells in too low a quantity to mimic
caloric restriction by more than a very modest degree. In these quantities, it
up regulates mitochondrial neogenesis more so than regenesis by a
moderate but significant up regulation of thyroxine. The purported
mechanism is via its mimicry of beta-estradiol activation of hypothalamic
stimulation of thyroxine stimulating hormone releasing hormone. But,
resveratrol is also a powerful anti-oxidant. In this mixed effect, it acts as a
caloric restriction meta-mimetic rather than an actual AMPK stimulating
mimetic. It does not significantly up regulate AMPK, but it activates PGC-
1alpha without up regulating TOR, so it is not acting like growth hormone,
either. This is a huge difference because, like CR elevation of AMPK, it is
definitely not anabolic and it sports its own catabolic influence. On the
other hand, it is unlike thyroxine stimulation in that it scavenges ROS,
evoking a regenesis rather than a neogenesis result, another net outcome of
CR. Regardless, resveratrol is more neogenic than regenic in these
quantities. How much its mixed CR mimetic function plays into this
tortuous scenario, is anybody’s guess, and its numerical impact on the ROS
load is unknown. The largest evidence supporting this scenario is that
dietary resveratrol’s cancer killing and life extension functions, as seen at
in vitro concentrations, some ten times higher, are not significantly
manifest. At the dietary levels it would be prudent to force a neogenic
default to regenesis just to hedge the bet toward a definitive CR like
outcome. Vigorous endurance exercise after an over night fast would
probably turn the trick via creating a sugar depletion oxygen debt. In
humans, resveratrol followed by exercise, even without fasting, converts
glycolytic fast twitch white muscle fibers into aerobic red slow twitch
fibers, and increases mitochondrial cell volume from about5% to 30%, with
regenic characteristics. Similar results occur in fasting runners in the
absence of resveratrol. The experiments we need are rather obvious.

The in vitro life extension and cancer apoptotic effects of resveratrol in the
20-50 uM range are so enticing that this dietary bioavailability deficiency
should soon be overcome. Already, free resveratrol serum levels well above
the 20-50 uM target range are being reported, industrially. Also, synergistic
molecules like quercetin and co enzyme Q with antioxidants are in the mix.
Still, TOR down regulation and mitochondrial regenesis are requirements
for full CR impact. We explore these issues in the practical applications
section, later. But, for now, our metabolic feedback loop attentions divert
their focus onto the cancer metabotype.

From a genetic perspective, cancer cells appear insane. Between the

chromosomal inversions, insertions and uneven cell divisions that render
cells into a heterogenous mix of hundreds to thousands of genes in
polyploidy or haploidy, add to this, the changed protein complement and its
post translational miss modifications, and the result becomes a bewildering
array of changes from any given cancer’s not quite fully differentiated stem
cell progenitors. Although each cancer cell type retains many of the
characteristics of its parent cell type, shot gun analysis, such as 2D
electrophoresis and tryptic digest multi-gradient HPLC (high performance
liquid chromatography) and MALDI-TOF (matrix assisted laser desorption
ionization-time of flight) mass spectrometry, demonstrate that no two
cancers are alike and that a given cancer is not even like itself. A cancer
tumor seems as if it is composed of a heterogenous thematic of a precursor
cell type engaged in a hyper-Darwinian selection for survival in an
organism that is desperately trying, but failing, to kill it.

Fortunately, even cancer cells must obey the laws of thermodynamics, and
they must do so within the constraints of the metabolic tool kit of sugar,
amino acid, lipid and nucleotide apportionments and redox (NADH/NAD)
and energy carriers (AMP/ADP/ATP) that permit the cell to grow and
divide, so as to be able to grow and divide, and so on. Note here, that the
sugar and carbohydrates are the cheapest energy source in the earth’s biotic
food web. Lipid is a bit pricier, but proteins and nucleotides are very
expensive because phosphate and redoxable nitrogen are nutrient limiting,
pretty much, planet wide. Thus, sugar is the preferred fuel during cell
growth and division (especially during hypoxia), while lipids, proteins and
nucleotides are preserved as building materials. These nutrient limitations
and metabolic requirements were fixed in stone a billion years before the
first multicellular organism existed. In addition, cancer cells share a lot of
features with fetal cells. Very importantly, they are always outrunning their
fuel and oxygen supply needs, so they take on a hypoxia metabotype that
induces blood vessels to grow toward them. This blood vessel growth is
called angiogenesis. They also dramatically increase their glycolytic rate, in
some cases well over 1000%. Most importantly of all, this changes the
glycolytic to mitochondrial ATP production ratio by even a greater
percentage, due to mitochondrial paucity and/or inefficiency. Like fetal
cells, they become glucose junkies as they parasitize the sugar making
hepatic gluconeogenic processes of their host, but unlike fetal cells, they
cannot turn glycolysis off.

Because of this hypoxia and its anabolic requirements, cancer cell growth is
always neogenically out pacing its regenic function. Many books have been
and still will be written about fetal and enviromental bioenergetics and its
evolutionary and organismal growth condition implications, and especially
now, even more so, that the primary elements of the CR pathway have been
clarified, and implicate cancer and other diseases of aging. But, for now,
the main points, described above, will suffice.

How all this CR based AMPK feedback loop stuff fits together to arrive at
cancer cell intermediary metabolism is an astonishing wonderment and a
stunning new achievement. We grant many laudits and heaps of praise for
the scientists who slogged through years and hundreds of thousands or even
millions of man hours to elucidate the particulars of the regulatory loop and
its myriad of attendant pathways. Even though cancer research and CR
authors (as well as diabetes and cardiovascular researchers) may not yet
know it, their results show that the cancer metabotype and the CR pathway
are the exact same pathway, albeit operating in basically opposite
directions and under different conditions, with the AMPK loop and
mitochondrial neogenesis vs. regenesis as core control elements in both
cases, and with cancer cell glycolysis providing its own caveat. The fact
that type II diabetes and cardiovascular hyperplasia and cardiac
hypertrophy are also outcomes of this pattern, is even more amazing.
Readers of this document would easily understand and be pleasantly
surprised by the basic cancer connection if they read (and this is a must
read) New Scientist, 5/15/2010 p.6, a brief outline, of which, is below, after
the Warburg correction discussion.

In the last two years, cancer researchers have proven that the particulars of
glycolytic and aerobic metabolism in cancer cells are precisely as
previously described in 1980. At that time the hypothesis corrected a flaw
in the Warburg aerobic glycolysis hypothesis by postulating that cancer cell
mitochondria suffered from an ATP production shortfall resulting in an
anaerobic to aerobic ATP differential as opposed to Warburg’s
mitochondrial oxygen consumption deficiency concept. This is an
important difference because it changes the NADH/NAD, ATP/ADP and
substrate flows in the cell. It was also described, in detail, how this shortfall
was integrated with glycolytic fetal enzyme activation to force the system
to amplify the, as then poorly understood, anabolic command system, to
operate in an irreversible state of cell growth and division, we previously
have coined as the cancer metabotype. Interesting methods of attack
designed to kill or renormalize cancer cells were envisioned at that time.
With our new found knowledge, we might wish to revisit some of these
strategies, in a search for synergies. It is remarkably serendipitous how the
‘rediscovery’ of this forgotten system during the last two years is so
similarly identical in such a wide array of particulars. In all cases, whether
as old discoveries, rediscoveries or new discoveries, the results
unambiguously show that the cancer metabotype is obligately and
inextricably intertwined with the CR pathway, and with mitochondrial over
neogenesis with a paucity of regenesis. It is this synthesis that is the really
new stuff.

As an overview, from a metabolic standpoint, most cancer cells are stuck in

mutationally up regulated fetal enzyme induced hyper glycolysis supported
by down regulated AMPK to TOR etc. driven mitochondrial neogenic ATP
production deficiency, in ways that the cell cannot recover from, as it can
in fetal cells, at least in normal in vivo circumstances, short of artificial
intervention. Under the influence of increased mitogen drivers and
decreased or dysfunctional cell growth suppressors, the cell is forced into
irrepressible and irreversible rounds of growth and division. We ignore
metastasis and other issues, here, as we are focused on the ancient
metabolic drive states of cancer cell induction and maintenance as opposed
to progression.

What the researchers in the New Scientist review article found was that
blocking fetal pyruvate kinase enzyme driven glycolysis with
dichloroacetate ‘reawakened’ (their words) mitochondrial regenesis (our
words), re-establishing normal glycolytic to mitochondrial ATP production
ratios and metabolite flow, and brought cancer cell growth to a virtual halt
in brain glioblastomas. Furthermore, previous hospital records showed that
up regulation of AMPK with metformin in diabetic lung cancer victims
increased survival times. This is the first time, in humans, for it to be
shown that CR and anti-cancer therapies share the exact same AMPK
control circuit, and with an anti-diabetes drug, to boot. We must also note
that the system has been shown to work in numerous mouse cancers, and
this is one time in cancer research that mice and men really share an ancient
and attackable metabolic control pathway. This is just the early
‘sledgehammer’ phase of the human work. More sophisticated assaults are
envisioned with excited expectancy. Let us hope that we don’t have to find
too many fetal enzyme blockers, because even thirty years ago, we knew of
other such fetal enzyme supporting changes in glycolysis and its attendant
anabolic NADPH providing pentose phosphate shunt, in different cancers.

Reducing hyperglycolysis and increasing mitochondrial efficiency are the

two key elements in slowing the impact of age related like cancer, type II
diabetes and cardiovascular dysfunction. For instance, in the aging adult,
cardiovascular ROS damage institutes mitogenic and hyperplasic
thickening in the intima of the vascular tree, and ventricular hypertrophy
concomitant with mitochondrial decline and a steady state shift from
efficient high energy lipid catabolism to low efficiency glucose catabolism.
Contrast this to hypoxic fetal conditions, when birth provides abundant
pulmonary oxygen followed by full cardiac mitochondrial biogenesis, a
rapid shift from carbohydrate to lipid catabolism, a conversion from
parasitic high affinity fetal hemoglobin F to free living lower affinity adult
hemoglobin A and a decrease in ROS production. The first is a stop-gap
response to ROS induced genetic damage, while the second is a finely
tuned genetic system response to an energy opportunity. Both are mediated
by the AMPK, TOR etc. feedback loop, and judicious stimulation of this
loop via CR or its mimetics are shown to yield significant inhibitory
impacts on these exact forms of cardiovascular decline. Type II diabetes is
a similar whole body response to the same type if insults found in
cardiovascular decay, and can be staved off by direct metformin activation
of AMPK that lowers insulin resistance and creates cellular housecleaning
and mitochondrial regenesis. Metformin has been in use from long before
we were even aware of such things as the sestrin feedback loop,
mitochondrial neogenesis, regenesis and its relation to fetal
hyperglycolysis!

Both neogenesis and regenesis are critical to retaining a juvenile youth state
in adult cells. By analogy, neogenesis is like a construction crew that builds
a new house but leaves a mess of construction related debris. Regenesis is
like the housecleaning and maintenance follow up that makes the house
into a livable home. In the aging adult non-dividing cell, both functions are
deficient with regenesis being the more deficient, as aging cells tend to
progress toward debris ridden hyperplasia. Declining growth hormone and
androgenous steroids inhibit neogenic and rejuvenative functions while
ROS exacerbates ‘rusting out’ functions and abundant nutrition inhibits
regenic functions. This may explain why CR seems to have virtually no
impact if started before early middle age. This also may explain how
chronic neogenesis, as with growth hormone, yields short term gains with
long term ‘burnout’. Conversely, it may explain how chronic regenesis, as
with CR, yields long term gains, but with serious quality issues. Perhaps
periodic neogenesis with a rapid default to regenesis, as is much more
typical of juvenile cells, may provide an optimal result.

A review of these mechanisms has left the two of these authors quite
breathless when we consider the scope of its medical implications, both in
terms of the medical monetary cost savings, and in terms of the staggering
number of quality human life years which might be added to the aggregate.
It is quite nifty how key elements of the three greatest killers of the human
race, along with a healthy squirt from the fountain of youth, all fit within
the framework of a singular, unifying and global hypothesis. Thus, from all
that has been articulated in the above document, we define the system.

PRACTICAL APPLICATIONS: THE FUTURE BURNS BRIGHT

One might consider this to be the wild hypothesis part of the document, but
based upon the aforementioned findings, the proposals might actually be
more sound than a lot of the stuff that peppers the grocery store checkout
stands. However we remind everyone that we are not medical doctors, and
therefore, not licensed to practice medicine. Nor is it our intention to do so.
Any use of the information contained in same is at the reader’s discretion.
We specifically disclaim any and all liability arising directly or indirectly
from the use or application of any information contained in any of these
articles. What we write here, is more so, of a free speculation of ideas
engaging over the counter phytonutrients and life style choices.

Recent longitudinal studies show us that dietary sugar is killing us more

resolutely than either saturated fat or high protein content, as found in
animal products. The diseases of aging, such as diabetes, heart disease and
cancer, kill the vast majority of us and excess dietary (extracellular) sugar
is a main and growing culprit. However, the previously outlined AMPK,
TOR, PGC-1alpha, ROS, SESN cycle shows that the system’s relentless
decay yields to the diseases of aging, in spite of dietary sugar, due to
intracellular metabolic shifts over time. In other words, even though sugar
powered the creation of our lives from inception to birth, it will eventually
kill us even if we don’t eat it. This is ‘natural’ aging, and the data clearly
shows that unnatural or supra-natural efforts must be made to obtain the
unnatural or supra-natural state called life extension beyond the natural or
normally expected limit. Face it, caloric restriction (CR) is draconian and is
only natural in the sense that, in nature, food sometimes runs out. No
organism exists that will ‘naturally’ CR itself in the presence of adequate
food. Mega doses of anti-oxidants or a hundred bottles of wine worth of
resveratrol a day is, decidedly, unnatural.

That being said, such things have been found to stand the test of time. For
instance, the Chinese have been drinking a high resveratrol Japanese
knotweed root extract, called itadoli tea, for millennia with claimed
beneficial results and no known ill effects save for some occasional
intestinal discomfort, found to be mostly due to emodin, a co extract, which
incidentally, is not found in modern concoctions. The remainder of this
brief discussion is mostly devoted to some unnaturally ‘natural’ stuff that
folks might do without having to live a supplement menagerie supported
life in near anorexia with its attendant impediments to muscularity, wound
healing, immune function, fecundity and others. The focus, here will be on
forcing a default state to mitochondrial regenesis, which is the heart and
soul of life extension and the inhibition of cancer cell induction and
maintenance. Thus, the discussion will completely avoid the well worn
school marm admonitions such as ‘eat your vegetables,’ ‘take your
vitamins,’ ‘exercise regularly,’ ‘drink plenty of water,’ ‘keep your weight
down,’ ‘don’t eat between meals,’ ‘brush your teeth after every meal,’
‘don’t eat anything that you can’t fit into your mouth,’ ‘holy cow, that sure
is a big fish’ and other common sense standard fare that we won’t even
mention, here.

It is difficult to tell how much of dietary resveratrol is neogenic and how

much is regenic. It cannot be heavily neogenic due to its TOR bypass,
catabolic and anti-oxidant functions plus its average life expectancy
increasing and rejuvenative outcomes in the absence of increased cancer
induction or any true life length reduction. Standard dietary resveratrol also
cannot be very strongly regenic because there is no appreciable up
regulation in AMPK, no real reduction in cancer incidence, no increase in
cancer cell apoptosis and no true life extension. However, much
mitochondrial biogenesis is observed, but how much of this is neogenesis
converted to regenesis is unknown because most investigators were
unaware of the difference.
Progressive nutriceutical supply companies recognize this and are actively
pursuing remedies. The essential problem is simple. When resveratrol is
eaten, well over 90% of it is sulfonated and glucuronidated in the intestines
and in the liver via the hepatic portal system, rendering it water soluble for
targeting it for kidney removal to the urinary tract. Free, unmodified dietary
resveratrol is typically less than 2 uM during its one and a half hour
elevated blood plasma phase following ingestion. Glucuronate and sulfate
derivatized resveratrol do not cross interstitial cell membranes and the low
free resveratrol in cell microsomal fractions indicate that the interstitial cell
extracellular sulfatases and glucuronidases have no general impact. This
may not be true at sites of inflammation, but the desired whole body effect
appears not to be there. Although we have no retention or turnover numbers
on free intracellular resveratrol, it does not appear to accumulate over time.
Experiments have shown that serum soluble free resveratrol concentration
in the 20 uM range definitely impact the CR/AMPK pathway. For instance,
DMSO solubilized resveratrol, when injected into mice, causes all the
desired AMPK and downstream effects in brain tissue.

As mentioned earlier, nutraceutical suppliers are being very ingenious in

their attempts to get serum resveratrol concentrations up to the CR mimetic
range. One supplier shows, graphically, how micronization of resveratrol
dramatically increases free resveratrol up to and well beyond the minimal
required AMPK activating range. Other suppliers are creating encapsulated,
solubilized and time released formats. One supplier provides lozenges for
sublingual delivery. Resveratrol mixed with synergizers such as quercetin
and co enzyme Q, are also offered. We eagerly await their time based
serum data, and even more so, their intracellular AMPK up regulation data.

Here’s a quick and dirty home remedy. You can dissolve up to about 500
mg of resveratrol in a shot of 86 proof booze, or flavored schnaaps for the
more faint of heart. Solubility is mostly dependant upon alcohol content, so
the same should hold for a glass of wine, but it may take a while to
dissolve. Besides, the smaller the volume, the better it is for non-intestinal
absorption. Take a slug, swirl it around in your mouth for a full minute
before swallowing, kick back, and enjoy. Dissolving is what you might call
the nanonization technique. It has been shown to enhance buccal and
aerodigestive absorption by as much as 800% above dietary methods, and it
does not increase the kidney metabolite load one whit. Anyway, many of
the medical gurus out there tell us that 10-20 grams of ethanol a day, is
good for us.

Then, there’s the stretch (or even strain) of your imagination beyond the
orbit of Pluto, plan. Human beings have daily circadian and monthly lunar
cycles for a reason. Anyone who has read about the circadian melatonin
cycle knows what we’re talking about. The fact that human females have a
lunar cycle length receptivity cycle, and that in small, tight knit groups,
cycle together, is no accident. The seasonal based cycle was replaced by the
lunar cycle because conditions made it happen. What made it happen, you
query? We never thought you would ask.

The mutational force is the primary evolutionary driver, but it is accidental,

generational and is usually a losing proposition, with rare selective
advantage. But, the winners support population survival in the form of
multi-generational adaptation. The selection advantage of a switch from a
seasonal to lunar cycle must have been powerful, simply because it was
forced into existence. Consider the following. Over 99% of the last two
million years, or so, of human evolution, has been devoted to the slow
steady conversion from gathering, to scavenger gathering, to hunter
gathering, while the remaining less than 1% is called civilization. Having
excellent 3-D color vision, but poor night vision, the night light of the full
moon became a great advantage as it advanced geographical food acquiring
range and easier pickings. Being bipedal and having prehensile dexterity in
a shrinking dryas ecosystem were great evolutionary pre-adaptations and
opportunity vs. death drivers for proto-humans.

This dynamic food energy switch created full moon super-nutrition

followed a remaining month of being trapped in standard fare. This tuned
up a monthly cycle of high, then haphazard nutrition, which caused
elevated body fat that could conveniently support fecundity, two weeks
after the full moon, during the pitch black nights of the new moon. Since
nobody looks ugly in the dark, and besides, there being nothing else to do
but stumble around like blind idiots, a nutritional match was made in
heaven. In addition, sexual glue is social glue. This was already operating
in the context of a proto-hominid large brained noisy critter, with its
ecological niche pushing a movement toward even more brain growth,
symbolic representation as language and a pre-civilization social tool kit
simply awaiting large enough population numbers to invent neat stuff like
cities, war and jacuzzies. Thus, the African stage was set, and fortunately
for us, all four acts played out before we, as the genetic evidence shows,
were almost extincted.

This pattern would also be reflected as a natural nutrient driven cycle of

low and high AMPK activity and a regular neogenesis and regenesis cycle,
which may have assisted us in becoming the longest living primate. The
long parturition period, the interminable time stretch from birth to sexual
maturity and the creation of history, in the need for elders to pass all that
big brained accumulation to the next generational batch of dull witted
dimbulbs that seem to arise with each generation, could have helped to
assist this life lengthening admixture. In the context of this paper, that is, if
there still is any context, this kind of long winded speculative wannabe has
just got to be followed with a circadian/lunar cycle recipe format.

The advent of commercially available high concentration bioavilable

resveratrol would open the door to possibilities that will really bring the
troops home. At full dose, it would bias the system toward true CR
mimicry of AMPK driven mitochondrial regenesis, while at one tenth full
dose, it would bias the system toward the neogenic meta-mimicry we
described in detail, before. Since neogenesis takes several days to
complete, while regenesis is much quicker, a circadian/lunar cycle plan that
forces the system into lengthy defaults to life extending regenesis, might
look something like this: Low dose neogenic resveratrol could be
coordinated with anti-oxidants, dietary nutrient loading and/or power based
exercise for five to seven days, or so, then followed by high dose
resveratrol coordinated with daily food avoidance between dinner and
breakfast and/or high oxygen utilizing endurance exercise prior to breakfast
in a more extended time frame, say about three weeks, to entrench
mitochondrial efficiency, cellular house cleaning and a shift away from
glucose toward fat burning. Here we have something for everybody.
Lounge lizards could reap the benefits of the phytonutrient pattern only
effect, while the more restless spirits among us could enhance those effects
by dietary and exercise regimens. Either way, organs and tissues other than
cardiovascular and skeletal muscle could become larger recipients of
resveratrol’s benefits. Numerous daily, weekly and monthly variations of
the theme could be envisioned. One plan might be to include one meal a
day to cause chronic fasting default to regenesis and/or with exercise to
assist regenesis with glucose nutrient debt and/or a CR mimetic to activate
the AMPK life extension loop.

A very interesting rat study of intermittent CR has put the world of CR

afficionados on its ear. Using alternate days of ad libitum food supply and
total fasting, in rats, results in no long term CR, as the rats make up for
fasting by feasting between fast days, while ending up with life extension
comparable to CR. This more or less jives with the down regulation of
PGC-1alpha to regenesis turn on time frame. In fact, this study is the actual
proof of principle, since life extension by CR can’t happen without it. This
also jives with our notion that, at least partial neogenesis followed by
regenesis, won’t hurt life extension, and you can avoid the misery and
downside risks of genuine CR.

An interesting question emerges here. Since regenesis is a CR long term

life extension holding pattern, how long is it good for? By this, we mean:
Once mitochondria become efficient, how long do they stay efficient before
they need restimulated to become efficient again? In other words, how
many days of feast can one ‘get away with’ before each day of fasting? If
regenesis is good for a week, then a one day a week fast is a small price to
pay. Not only that, if 18 hours of fasting works as good as 24, then all one
would needs do is miss breakfast once a week. Can this be supplemented
with a low dose resveratrol regimen that would accentuate the effect?
We need experiments and we need them earlier rather than later. But rats
live five or more years, and rats are not people, (as opposed, off-times, to
the other way around). Over the counter metformin is over a decade away,
if ever, and rapamycin is too dangerous to become street legal in any time,
dimension or reality. Metformin, when used with growth hormone to
mechanistically force the system to toggle back and forth between
neogenesis and regenesis could ultimately turn out to be the greatest anti-
aging plus rejuvenation achievement of all time. This could truly cause
cells to behave more like they do in the juvenile stage. The timing, dosage
and testing required in such a scenario would be critical. This is playing
with some real big mojo, here, and it would be illegal without a
prescription under a doctor’s care. Only people, at least in their early
middle years, say past thirty, could participate in such a program, safely.

Fortunately, at present, we have resveratrol, which is freely obtainable and

we know enough about CR and mitochondrial neogenesis and regenesis, to
get answers, fast, and in humans instead of rats. Human volunteers and
tissue biopsies that measure CR via AMPK activity and mitochondrial
biogenic state from krebs cycle enzymes vs. cytochrome content, can allow
us to follow the system status through time. The system is well enough
defined, by now that the meanings of these assays point to causation rather
than correlation. A wide array of experiments could be rapidly conducted,
and could pinpoint which timings, conditions and regimens are optimal,
whether there are any down sides and what other items might be included.
Ideally, we may find a CR mimetic dose schedule of resveratrol, a better
mimetic or a mixture of synergistic components that require no life style
changes outside of normal prudent health practices … except for that magic
pill, of course.

AFTERWORD

Not very surprisingly, there is very little cross talk between the cancer
metabolism, cardiovascular disease, diabetes, life extension and
diet/exercise research communities. Today’s specialization doesn’t really
allow for it, and the areas of work do seem far afield of one another. But
there are other reasons. For one, the cancer metabolism field, although
rather ancient, is perceived to be actually, new. This has much to do with
the hugely bitter and public battle that led to Warburg’s demise in 1956.
The whole field of research became verboten, and one liners like “that was
proven false a long time ago” and “we don’t even look into that area any
more” were almost sheep mentality mantras. Another reason is that, who
would have ever guessed that any such connection between cancer and life
extension could even possibly exist, and even if it did, that it would
simultaneously lead to the fountain of youth and the slaying of the monster
hiding under the bed. Throwing in cardiovascular disease and diabetes, to
boot, is more icing than any cake can stand. But in reality, it isn’t too big to
believe, because this is all soon to become common knowledge. Anyone
who reads the three review papers referred to in this article can readily
observe that it has become too obvious by now for the connections not to
be seen by droves of scientists. And we don’t mean the small connections.
We mean the big ones, the really big ones.

Gregory S. Bambeck Ph.D. e-mail: gregorybambeck@yahoo.com

Michael Wolfson J.D., M.B.A. e-mail: mwolfson@stanfordalumni.org

Copyright © by Gregory Bambeck and Michael Wolfson June 11, 2010.