Aging and Potential Aging and Potential Interventions to Expand Interventions to Expand

LifespanLifespan

Ryan Shelton, NDRyan Shelton, ND

Longevity ProjectLongevity Project

• Continually involved, productive, Continually involved, productive, engaged in challenging interesting engaged in challenging interesting work (even in stress)work (even in stress)

• Being prudent, conscientious, Being prudent, conscientious, intentionalintentional

• Being socially involved and helping Being socially involved and helping othersothers

• Marriage for menMarriage for men

Biomarkers of Longevity in Biomarkers of Longevity in PrimatesPrimates

• TemperatureTemperature

• InsulinInsulin

• % of initial DHEA% of initial DHEA

• Calorie restrictionCalorie restriction– 6 mos, 48 humans VLCD reduced fasting 6 mos, 48 humans VLCD reduced fasting

insulin, body temperature, DNA insulin, body temperature, DNA fragmentationfragmentation

Definitions of agingDefinitions of aging• Accumulation of changes over timeAccumulation of changes over time

– Manifests as decline in functional and responsive Manifests as decline in functional and responsive capacity capacity

• Universal aging/Probabilistic agingUniversal aging/Probabilistic aging• Proximal aging/Distal agingProximal aging/Distal aging• Cellular senescence/Organismal senescenceCellular senescence/Organismal senescence

– Time of cell life vs # of cellular divisionsTime of cell life vs # of cellular divisions– Not known how cellular senescence plays a role in Not known how cellular senescence plays a role in

organismal senescence organismal senescence • Biological immortalityBiological immortality

– Stable rate of mortality as a function of chronological Stable rate of mortality as a function of chronological ageage

– Death requires injury or disease rather than Death requires injury or disease rather than deteriorationdeterioration

– Not an unavoidable property of life, rather part of the Not an unavoidable property of life, rather part of the genetic program genetic program

• Biogerontology (Biomedical gerontology)Biogerontology (Biomedical gerontology)

Theories of agingTheories of aging• Most evidence is correlative, not causative; all are Most evidence is correlative, not causative; all are

limitedlimited• Telomere shorteningTelomere shortening• Reproductive hormone theoryReproductive hormone theory• Damage/Error accumulation to genetic or Damage/Error accumulation to genetic or

epigenetic factorsepigenetic factors– Free radical theoryFree radical theory– Infections and inflammationInfections and inflammation– Micro-nutrient triageMicro-nutrient triage

• Mis-repair theoryMis-repair theory• MitohormesisMitohormesis• AGE cross linkageAGE cross linkage• EvolutionaryEvolutionary• Loss of circadian rhythmsLoss of circadian rhythms• Law of entropyLaw of entropy

Accelerated aging diseasesAccelerated aging diseases• ProgeriaProgeria

– Genetic mutation causing unstable Lamin A, part of the Genetic mutation causing unstable Lamin A, part of the building block of the nuclear envelope/scaffolding building block of the nuclear envelope/scaffolding

– Similar nuclear blebbing, nuclear receptor down-Similar nuclear blebbing, nuclear receptor down-regulation and DNA damage, poor DNA repair seen in regulation and DNA damage, poor DNA repair seen in the elderlythe elderly

– Patients do not show neurodegeneration or cancer Patients do not show neurodegeneration or cancer predispositionpredisposition

– ““Wear and tear” effects of aging not seenWear and tear” effects of aging not seen• Cataracts, osteoarthritisCataracts, osteoarthritis

• Segmental progeriasSegmental progerias– Dyskeratosis cogenitaDyskeratosis cogenita

• Poor telomere maintenancePoor telomere maintenance• DNA helicase mutationsDNA helicase mutations

– Werner syndromeWerner syndrome• Increased telomere attritionIncreased telomere attrition• DNA helicase mutationsDNA helicase mutations

Hayflick LimitHayflick Limit

• The number of times a cell population will The number of times a cell population will divide before it stops (Replicative senenscence)divide before it stops (Replicative senenscence)

• Human cells around 50-60 divisionsHuman cells around 50-60 divisions– 70 divisions at physiological O70 divisions at physiological O22 conditions conditions– Tortoise 100 divisions, Mice 15 Tortoise 100 divisions, Mice 15

• May have been selected to prevent cancerMay have been selected to prevent cancer

• Only Only sometimes sometimes correlates with telomere correlates with telomere lengthlength

• Does not address GDoes not address G0 0 phase of cell cyclephase of cell cycle

• In vivoIn vivo Hayflick Limit does not hold Hayflick Limit does not hold

ApoptosisApoptosis

• Programmed cell Programmed cell deathdeath

• 60 billion cells in 60 billion cells in human adult, 30 human adult, 30 billion cells in childbillion cells in child

• Apoptosis, Apoptosis, senescence, senescence, quiescencequiescence

• Atrophic factorsAtrophic factors– Diminished Diminished

innervation, innervation, blood supply, blood supply, nutrition, nutrition, endocrine endocrine stimulation; stimulation; toxins, infections. toxins, infections. senilitysenility

AutophagyAutophagy

• Degradation of intracellular proteins, Degradation of intracellular proteins, organelles, membranes as mechanism of organelles, membranes as mechanism of repair maintenancerepair maintenance

• Taking out the trash or recyclingTaking out the trash or recycling

• Stimulated by nutrient deficiency or Stimulated by nutrient deficiency or hypoxiahypoxia

• Slows with ageSlows with age

• Increase in autophagy associated with Increase in autophagy associated with longer life span in worms, flies, and micelonger life span in worms, flies, and mice

Important Genes/EnzymesImportant Genes/Enzymes• Sirtuin familySirtuin family

– NAD+ dependent protein deacetylase enzymeNAD+ dependent protein deacetylase enzyme– Regulates metabolism, transcription, stress resistanceRegulates metabolism, transcription, stress resistance– Tighter packing of chromatin, silencing most prominent at telomeric Tighter packing of chromatin, silencing most prominent at telomeric

and rDNA sequences; prevents early cell division, preserves replicationand rDNA sequences; prevents early cell division, preserves replication– Overexpression 30% increase in lifespan, deletion 50% decrease Overexpression 30% increase in lifespan, deletion 50% decrease

lifespan and eliminates life extension of calorie restrictionlifespan and eliminates life extension of calorie restriction– Stimulates autophagy, decreases mating factors, telomere Stimulates autophagy, decreases mating factors, telomere

maintenancemaintenance– Attenuates NF-kb signalingAttenuates NF-kb signaling

• mTOR enzymemTOR enzyme– Regulates cell growth, proliferation, autophagyRegulates cell growth, proliferation, autophagy– Decreased TOR activity slows agingDecreased TOR activity slows aging– Decrease mitogenic signaling, immunosuppressant, cancer TxDecrease mitogenic signaling, immunosuppressant, cancer Tx

• Daf-2, Daf-16 (Forkhead/FOXO)Daf-2, Daf-16 (Forkhead/FOXO)– Insulin/IGF-1 like receptor genesInsulin/IGF-1 like receptor genes– Decrease signaling increases life-span analogous to calorie restrictionDecrease signaling increases life-span analogous to calorie restriction– Repression in early life important for later lifeRepression in early life important for later life

• TelomeraseTelomerase– Only expressed in stem cells, germ cells, certain WBCsOnly expressed in stem cells, germ cells, certain WBCs– Increased life span, concern with cancerIncreased life span, concern with cancer

• AMP KinaseAMP Kinase• Matrix MetalloproteinasesMatrix Metalloproteinases• RAGERAGE

– Activate NF-kb, PRO-inflammatoryActivate NF-kb, PRO-inflammatory– Receptor for AGEReceptor for AGE

Important Genes/EnzymesImportant Genes/Enzymes

• HIGHLY CONSERVED ENZYMESHIGHLY CONSERVED ENZYMES• Sirt, DAF, TOR AMP kinases each shift away from Sirt, DAF, TOR AMP kinases each shift away from

growth and reproduction towards protection and growth and reproduction towards protection and maintenance VIA DIFFERENT PATHWAYSmaintenance VIA DIFFERENT PATHWAYS

• PEPCK-CPEPCK-C– Overexpression 50% life extension in miceOverexpression 50% life extension in mice

• PPAR receptorPPAR receptor– Deletion 30% life extension in miceDeletion 30% life extension in mice

• PGC-1 alphaPGC-1 alpha– Master integrator of external stimuliMaster integrator of external stimuli

• P53P53– Guardian of genome repair mechanismsGuardian of genome repair mechanisms– ‘‘Tumor suppressor gene’Tumor suppressor gene’– Longer life vs cancerLonger life vs cancer

TelomereTelomere

Telomere TheoryTelomere Theory• Protective ends of DNA, disposable buffersProtective ends of DNA, disposable buffers

• Biological clock or fuseBiological clock or fuse

• Repetitive sequence, does not code for proteinRepetitive sequence, does not code for protein

• Necessarily shortened with each cell mitotic divisionNecessarily shortened with each cell mitotic division

• Centenarians have longer telomeresCentenarians have longer telomeres

• Psychological stress negatively correlated with telomere length and Psychological stress negatively correlated with telomere length and telomerase activity; pessimism decreases WBC telomere lengthtelomerase activity; pessimism decreases WBC telomere length

• Shortened telomeres associated with a wide range of degenerative Shortened telomeres associated with a wide range of degenerative diseasesdiseases– Age, smokers, insulin, coronary artery calcium & atherosclerotic plaquesAge, smokers, insulin, coronary artery calcium & atherosclerotic plaques

• Telomeres highly susceptible to oxidative stress and are impacted Telomeres highly susceptible to oxidative stress and are impacted more by this than the end-replication problemmore by this than the end-replication problem

• Does not address GDoes not address G00 cell phase cell phase

• Mice lacking telomerase do not show decreased lifespanMice lacking telomerase do not show decreased lifespan

• Increase life span in mice seen only if genetically resistant to cancer Increase life span in mice seen only if genetically resistant to cancer

• Aged stem cells have decreased functional capacity and less Aged stem cells have decreased functional capacity and less functional lineagesfunctional lineages

Reproductive hormone Reproductive hormone theorytheory• Reproductive hormones may act in an antagonistic Reproductive hormones may act in an antagonistic

pleiotropic manner via cell cycle signalingpleiotropic manner via cell cycle signaling• Promotes growth and development early in life to Promotes growth and development early in life to

achieve reproductionachieve reproduction– Longer time to reproduce, longer life spanLonger time to reproduce, longer life span– Removal germline stem cells doubles lifespanRemoval germline stem cells doubles lifespan

• Decline and desynchronization drive senescenceDecline and desynchronization drive senescence• Young ovaries in old rats increases lifespanYoung ovaries in old rats increases lifespan• Longevity interventions decrease fertility by Longevity interventions decrease fertility by

decreasing HPG axisdecreasing HPG axis• HPG hormones affect longevity regulating pathwaysHPG hormones affect longevity regulating pathways• More telomerase activity with estrogen, testosterone, More telomerase activity with estrogen, testosterone,

DHEADHEA• Mice w/o pituitary gland on HRT lived longer than Mice w/o pituitary gland on HRT lived longer than

controlscontrols• Hypothalamus and pituitary maintain capacity, end Hypothalamus and pituitary maintain capacity, end

glands do notglands do not• Incomplete theoryIncomplete theory

Damage/Error accumulationDamage/Error accumulation

• VirusesViruses

Damage/Error AccumulationDamage/Error Accumulation

• 10,000 to one million lesions per cell per day10,000 to one million lesions per cell per day• Can alter/eliminate transcriptionCan alter/eliminate transcription• Damage vs mutationDamage vs mutation

– DNA damage special problem in slow or non-dividing cells: DNA damage special problem in slow or non-dividing cells: AgingAging

– DNA damage in fast dividing cells can cause mutation: DNA damage in fast dividing cells can cause mutation: death of cell or cancerdeath of cell or cancer

• Genes influential on lifespan involved in DNA repairGenes influential on lifespan involved in DNA repair• nDNA & mDNAnDNA & mDNA• EpigeneticsEpigenetics

– Non-genetic mechanisms of temporal and spatial genetic Non-genetic mechanisms of temporal and spatial genetic controlcontrol

– Methyl groups, histone tails, chromatin remodelingMethyl groups, histone tails, chromatin remodeling– Morphogens give cells memories by governing tissue Morphogens give cells memories by governing tissue

developmentdevelopment

Mis-repair TheoryMis-repair Theory

• Mis-repair was selected for as a Mis-repair was selected for as a survival advantagesurvival advantage

• Quick repair increases survival Quick repair increases survival chances to reproductionchances to reproduction

• Mis-repair accumulates over timeMis-repair accumulates over time• Distorted gene products less than Distorted gene products less than

optimal functionoptimal function• Aging is the price of survivalAging is the price of survival

MitohormesisMitohormesis

• Energy centers produce free radicalsEnergy centers produce free radicals• Induction of stress resistance through Induction of stress resistance through

mitochondrial anti-oxidantsmitochondrial anti-oxidants• With calorie restriction, there are more anti-With calorie restriction, there are more anti-

oxidants than free radicalsoxidants than free radicals• Anti-oxidant supplementation can Anti-oxidant supplementation can

downregulate this phenom to increase downregulate this phenom to increase morbitity/mortalitymorbitity/mortality

• Loss of redox control progresses cell cycle Loss of redox control progresses cell cycle from Gfrom G00 to proliferation or apoptosis to proliferation or apoptosis

• Aging associated with less anti-oxidant Aging associated with less anti-oxidant capacitycapacity

DNA Damage/ErrorDNA Damage/Error

AGE cross-linkageAGE cross-linkage

• Cooking ourselves to deathCooking ourselves to death– Reduces elasticity (1Reduces elasticity (1stst known effects of aging) known effects of aging)

• Externally, a non-enzymatic reaction between an amino Externally, a non-enzymatic reaction between an amino acid and a reducing sugar (browning reaction)acid and a reducing sugar (browning reaction)

• Internally, Advanced Glycation Endproducts undergo Internally, Advanced Glycation Endproducts undergo Amadori rearrangements to form ketosamines on nucleic Amadori rearrangements to form ketosamines on nucleic acids, proteins, and lipidsacids, proteins, and lipids– Form azomethine bondsForm azomethine bonds– Increased by oxidative stress, inflammation, hyperglycemia, Increased by oxidative stress, inflammation, hyperglycemia,

fructose intakefructose intake– Implicated in DM, CVD, Alzheimer’s, asthma, arthritis, Implicated in DM, CVD, Alzheimer’s, asthma, arthritis,

nephropathy, retinopathy, neuropathynephropathy, retinopathy, neuropathy– Cross-linking can effect membrane fluidity/transport, cell/cell Cross-linking can effect membrane fluidity/transport, cell/cell

communication, DNA damage, elasticity communication, DNA damage, elasticity – Interaction between AGE and RAGE highly pro-inflammatory Interaction between AGE and RAGE highly pro-inflammatory

and NF-kb upregulates more RAGE expression and NF-kb upregulates more RAGE expression – AGE induces Matrix MetalloproteasesAGE induces Matrix Metalloproteases

EvolutionaryEvolutionary• Aging is relatively new phenomenonAging is relatively new phenomenon• Little selection pressure in old ageLittle selection pressure in old age• Detrimental mutations more likely to show up later in Detrimental mutations more likely to show up later in

lifelife• Regulation of genetic expression program controlled by Regulation of genetic expression program controlled by

activating different genes at different times or growth, activating different genes at different times or growth, development, and day-today lifedevelopment, and day-today life

• Reproduction is king; lean years when reproduction was Reproduction is king; lean years when reproduction was less likely the growth and reproductive genes turned off less likely the growth and reproductive genes turned off in favor of protection and maintenance for a future in favor of protection and maintenance for a future opportunityopportunity

• Exceptions to the ruleExceptions to the rule– Naked mole ratNaked mole rat– TurtlesTurtles– Hydras, TardigradesHydras, Tardigrades– Bristlecone pinesBristlecone pines– Sturgeon, rockfish, lobsters, clamSturgeon, rockfish, lobsters, clam– Bowhead whaleBowhead whale

Possible interventionsPossible interventions

• From Gilgamesh to $50bill/yrFrom Gilgamesh to $50bill/yr• Most research has been Most research has been in vitroin vitro and and

non-mammal non-mammal in vivo in vivo studies studies • Best chances for drastic expansions Best chances for drastic expansions

in life span likely with nanomedicine, in life span likely with nanomedicine, gene therapies, stem cell therapiesgene therapies, stem cell therapies– Catalase gene therapy into mitochondria Catalase gene therapy into mitochondria

increase lifespan by 25% in miceincrease lifespan by 25% in mice

Possible interventionsPossible interventions

• Sleep (optimal) and maybe nappingSleep (optimal) and maybe napping• DietDiet

– Whey proteinWhey protein– NeolithicNeolithic

• No grain, dairy, processed foodsNo grain, dairy, processed foods– Fructose and sucroseFructose and sucrose

– Calorie restrictionCalorie restriction• Intermittent longer increase (50%) than constant restriction Intermittent longer increase (50%) than constant restriction

(30%)(30%)• 30-50% less than 30-50% less than ad libitumad libitum• Methionine restriction also worksMethionine restriction also works

• ExerciseExercise– Moderate, not excessive, and must be enjoyedModerate, not excessive, and must be enjoyed

TelomeraseTelomerase• Cancer and telomerase; p53 & Sirtuin?Cancer and telomerase; p53 & Sirtuin?• Studies in mice show rejuvenation of aged tissueStudies in mice show rejuvenation of aged tissue• Telomerase decreaseTelomerase decrease

– Psychological stressPsychological stress– Oxidative stressOxidative stress– Caution with plant polyphenols?Caution with plant polyphenols?

• Telomerase increaseTelomerase increase– EstradiolEstradiol– TamoxifenTamoxifen– Pioglitazone (Actos)Pioglitazone (Actos)– GHRH antagonistGHRH antagonist– Erythropoietin Erythropoietin – TA-65 (Astragalus extract)TA-65 (Astragalus extract)– Terminalia chebulaTerminalia chebula– Portulaca oleraceaPortulaca oleracea– CistancheCistanche– Colostrum (Epidermal growth factor)Colostrum (Epidermal growth factor)– MelatoninMelatonin– Reduction of IL-6Reduction of IL-6

SirtuinSirtuin

• Are hypothalamic Sirt enzymes most important?Are hypothalamic Sirt enzymes most important?• Pharmacological mimicking of calorie restriction elicits Pharmacological mimicking of calorie restriction elicits

epigenetic reprogramming of differentiated cells to more epigenetic reprogramming of differentiated cells to more pluripotent self-renewal statespluripotent self-renewal states

• Resveratrol, PiceatannolResveratrol, Piceatannol– SRT 1460, SRT 2183, SRT 1720SRT 1460, SRT 2183, SRT 1720

• PterostilbenePterostilbene• Orsirtine GLOrsirtine GL• Fisetin, ButeinFisetin, Butein• Stabilized oxaloacetateStabilized oxaloacetate

– Also may work on FOXO & AMP kinase pathways through Also may work on FOXO & AMP kinase pathways through increasing NAD+/NADH ratioincreasing NAD+/NADH ratio

• MelatoninMelatonin– Also reduces body tempAlso reduces body temp

TOR enzymeTOR enzyme

• Inhibited byInhibited by– Reduced calorie intakeReduced calorie intake– CaffeineCaffeine– RapamycinRapamycin– CurcuminCurcumin– EGCGEGCG– ResveratrolResveratrol

AGE BlockadeAGE Blockade

• L-CarnosineL-Carnosine• BenfotiamineBenfotiamine• Aminoguanidine, MetforminAminoguanidine, Metformin• Butein, Cinnamon, clove, cherry blossom flowers, Butein, Cinnamon, clove, cherry blossom flowers,

Terminalia, red yeast rice, grape seed extract, Terminalia, red yeast rice, grape seed extract, Sophora, Withania, ZeaSophora, Withania, Zea

• AdiponectinAdiponectin• Genistein, ECGCGenistein, ECGC• Phloridzin, phloretinPhloridzin, phloretin• AlagebriumAlagebrium• Acetylsalicylic acidAcetylsalicylic acid

Random mechanismsRandom mechanisms• Berberine (reduce MMP)Berberine (reduce MMP)• 2-deoxyglucose mimics calorie restriction2-deoxyglucose mimics calorie restriction• Methylators are generally gene silencersMethylators are generally gene silencers

– 5-THF, TMG, MethylB12, B65-THF, TMG, MethylB12, B6• SelegilineSelegiline

– Increase SOD?Increase SOD?• Non-TOR induced autophagyNon-TOR induced autophagy

– Clonidine, verapamil, spermidine, minoxidilClonidine, verapamil, spermidine, minoxidil• Chinese herbsChinese herbs

– Fo-Ti, Lycium, Panax, RehmanniaFo-Ti, Lycium, Panax, Rehmannia• Mitochondrial antioxidantsMitochondrial antioxidants

– Alpha lipoic acidAlpha lipoic acid– Acetyl-L-carnitineAcetyl-L-carnitine– N-acetylcysteineN-acetylcysteine

• GGT-knockout mice develop cataracts in 3 weeks, die in 12 weeksGGT-knockout mice develop cataracts in 3 weeks, die in 12 weeks– TaurineTaurine– SkQSkQ– CoQ10CoQ10– TempolTempol– CatalpolCatalpol– MitoQ, MitoEMitoQ, MitoE

• Alkylated triphenylphosphonium to CoQ10 & Vit EAlkylated triphenylphosphonium to CoQ10 & Vit E

Unknown mechanisms for Unknown mechanisms for micemice• Vit DVit D• B5B5• Aged garlicAged garlic• DHEADHEA• Ganodermasides, CordycepsGanodermasides, Cordyceps• Nucleic acidsNucleic acids• CurcuminCurcumin• MetforminMetformin

– Especially females, early treatmentEspecially females, early treatment

BHRTBHRT

• Optimize bio-identical hormonesOptimize bio-identical hormones

• Consider DHEAConsider DHEA

• HRT patients still ageHRT patients still age

• Circadian pulsatility may be importantCircadian pulsatility may be important– SIRT1 actually involved in circadian SIRT1 actually involved in circadian

rhythmsrhythms

• GH, IGF-1 may decrease life spanGH, IGF-1 may decrease life span

Important concernsImportant concerns• Cell immortality induces oncogenes or lose tumor Cell immortality induces oncogenes or lose tumor

suppressor genessuppressor genes• Life span extension verses quality improvement and Life span extension verses quality improvement and

disease reduction; Optimize natural longevitydisease reduction; Optimize natural longevity• Aging is variable, aging accumulates with ageAging is variable, aging accumulates with age• No proven method in humans of life span expansion; No proven method in humans of life span expansion;

medical focus is on prevention and early detection of medical focus is on prevention and early detection of diseasedisease

• As a system advances through time, it will statistically As a system advances through time, it will statistically become more disordered and living systems decrease become more disordered and living systems decrease their entropy by expenditure of energy at the expense at their entropy by expenditure of energy at the expense at the expense of environmental entropy increasethe expense of environmental entropy increase

• For every measurable value, the variation in the For every measurable value, the variation in the distribution in values increases with agedistribution in values increases with age

• Can GGT be a marker for disease, oxidative stress, and Can GGT be a marker for disease, oxidative stress, and aging?aging?

• Cool Database: http://human.ageing-map.org/Cool Database: http://human.ageing-map.org/