HUMANAGEING

ANDLONGEVITY

ESOF 2006Munich, 16 July 2006

Claudio Franceschi

CIGInterdepartmental Centre

“L. Galvani”University of Bologna

ITALY

The highest national life expectancyobserved for female, 1580 - present

ASPETTATIVA

DI

VITA

Life expectancy doubled in the last two centuries(data refer to women)

Max Plank Institute for Demography, Rostock, Germany, Annual Report 2001

In the last 50 years the mortalityof people over 80 years decreased dramatically

Rectangularization and extensionof survival curves

What changed around 1950?

Maybe a change of attitude?

Vitamins?Antibiotics?Diet?Exercise?Progress of gerontology?All of the above?

Yashin et al., Exp Gerontol (2001) 37, 157-167

In some part of the world(rich, developed countries)

we have optimizedthe environment

(nutrition, hygene, medicine, housing and working

conditions)

After age 80the rate of mortality

decreasesand is no more

describedaccurately by the

Gompertz’s equation

centenarians

In the last 50 years the number ofcentenarians increased

in a very fast way(they double every 5-7 years)

Centenarians still representan extreme phenotype: 1:8-10,000

The oldest person in the world

Demographythe Sardinia island as a

laboratory

Table 1. Comparative figures on centenarian prevalence and sex ratio (source : Statistical Offices of respective countries) Countries and Regions Date Prevalence

(number of living centenarians per 100.000 inhabitants)

Sex ratio (among living centenarians females/males)

Belgium January 1st 2002 10.5 7.08 Denmark July 1st 2002 10.4 6.27 Sweden January 1st 2003 12.6 5.64 Italy January 1st 2001 14.1 3.80 Sardinia January 1st 2001 16.6 2.70 Nuoro province January 1st 2001 17.9 1.40

In Italy the number of centenarians is7200 (2004) out of 57.5 M inhabitants

In Italy there is a North to South gradient in the female:male ratio in centenarians. Male centenarians are more frequent in the South

BOLOGNA

Passarino et al., Exp. Gerontol. 37, 1283-1289, 2002

Male centenarians are particularly frequent in Sardinia

Passarino et al., Exp. Gerontol. 37, 1283-1289, 2002

Figure 1. Mapping significant positive differences between observed number of centenarians and theoretical numbers based of average Sardinian figure with Gaussian smoothing method and 15 km distance.

Cagliari

Sassari

Iglesias

Alghero Ozieri

Oristano

Quartu

Tempio

Carloforte

BosaBonorva

Guspini

Nuoro

Sorso

Ittiri

Olbia

0km 25km 50km

Average number ofcentenarians for

1000 births

Khi-square test of areaswith significant positive

deviations to meanvalue of Sardinia

5 %1 %0.1 %

Grasland C., 2003, The Hypercarte Project

0.2

1.2

2.2

3.2

4.2

Mean value of Sardinia

Centenarians are not randomly distributed(according the their place of birth)

A geographic area highly enriched in male centenarians has been identifiedin Sardinia

The Blue Zone

Deiana et al., Exp. Gerontol., 39:1423-9, 2004

A demographic-epidemiologicalanalysis

is revealing that the remarkable ratio between women and men in Sardinia is

related to a lower mortality of men, particularly for CVD,

after age 80

Caselli et al., in preparation

Peculiar combination of genetics and lifestyle ?

Longevity = Environment + Genetics + StocasticityL=E+G+S

Antonio Todde enjoying his 112th birthday

77

Todde E

75

Todde G

73

Todde A

70

Todde G

59

Todde G

63

Todde M

75

Todde A Deiana MM

103

93

Deiana GA

79

DeianaMCA

Todde AD

78

81

Deiana A

63

Deiana F Deiana M

69

Deiana G

76

59

Todde MA

3Todde AD

98Todde A.R Todde A.

Todde L

82 77

Todde G

63

Todde ATodde A

79 1

Todde I

112

Todde Antonio Madeddu MA

86

98

Deiana FA Todde FM

88

sb

still birthTodde G

30 90

Madeddu B Todde G

70

Todde S

45

Todde A

72 101

Todde MA Todde A

72

Todde FA

60

37

Noli MRDeiana AM

46 38

Noli MESTodde AS

87

87

Lai GNoli GBDeiana GL

78 76

Murru C

Murru C Todde MA Lai G Lai M

Todde S

75

Fenude A

Todde G

~40 41

Mattu A

Todde A Lai C Mattu R

~50

57

Todde SNoli G Matzone S

Deiana M

§

§ isonymic marriage

twins

Family tree of Antonio Todde (1889-2002) The number inside the symbols (circle for women and square for men)

represents the age in years at death (symbols grey) and the age reached (symbols white)

Caselli et. Al., Exp Gerontol 2006

The maternal lineage is enriched in nonagenariansand centenarians

Genetics and Longevity

• Parents of centenarians lived longer thanpeople of the same cohort

• Siblings of centenarians have a “risk” toreach 100 several times higher than thatof people of the same cohort

• Offspring of centenarians have a lowermortality and are protected from CVD and cancer

• A strong familiar component of longevity

Genetics and LongevityThe survival advantage of centenarianoffspring is not shared by their spousesdespite the fact that they shared the sameenvironment for most of their life

Thus the strong familiar component of longevity is likely a genetic componentand long living sibs should be highlyenriched in longevity genes

L=E+G+SLongevity = Environment + Genetics + Stocasticity

The equation of longevity

Longevity is a very complex trait

Longevity genes in humans (association studies) STRESS RESPONSE

HFE TH IFNγ IGF1-R HRAS1

FibrinogenFactor VIIFactor VPAI-1

RISK FACTORS FOR CARDIOVASCULAR DISEASES,DEMENTIA AND DIABETES

APOBAPOA1APOC3APOA4

HSP70 TP53PARP

WAF-1Y CHROM

CANCER, DNA REPAIR, CELLULAR PROLIFERATION, APOPTOSIS

TNFα IL6TLR4 Pyrin

IL-1β CD95IL-1α IL-1RADefensins

INFLAMMATION-Immune response

RENINSTPO

SOD1SOD2

IL10 TGFβ1

SIRT3with longevity

In red and bold gene-positive associations

FAS

PON1 APOE

PPARγ

GEHAGEnetics of Healthy Aging

Integrated Project of EU 6thFP7.2 M €, 25 Units

(coordinator C. Franceschi)Recruitment and Genome Scanning(nuclear and mitochondrial genomes)

of 2650 90+ sibpairs collected in 12 countriesStarting date: May 1st 2004

It is possible to becamecentenarians

by- different strategies

- different combinations of E,G,S

The unusual genetics of longevity betweenantagonist pleiotropy and adaptive physiological

remodelling

AdaptationAdaptationRemodellingRemodelling

AntagonistAntagonistPleiotropyPleiotropy

LONGEVITY = Capability of the body

1.to cope with stressors2. to remember the experience of stressors

and thus3. to adapt and to remodel (hormesis?)

at the molecular, cellular and organismallevel (psychological, immunological)

the consequence is a remarkable change of the internal environment and “context”

- This process is quite variable from individual to individual and it is under genetic control which likely increases with age

the model of centenarians

centenarians are likelynot simply the more robust

butthose people who adapted

and remodelled

better and quicker(more hormetic ?)

remodellingselection +Centenarians represent

an extreme phenotype: 1:8-10,000

STRESSORS

MAINTENANCE SYSTEMS(Defence and Repair)

ADAPTATION

REMODELLING DISEASE

LONGEVITY DEATH

STRESSORS

MAINTENANCE SYSTEMS(Defence and Repair)

Change in body microenvironment

Changes in geneexpression

Progressive accumulation of “scars”

Changes in proteinabundance, composition

interaction

THE AGE-ASSOCIATEDREMODELLINGIS ASSOCIATED WITH A LOSS OF

COMPLEXITY

Reduction in physiologic inputs, loss of complexity and functional level

Lipsitz, Sci Aging Knowl Environ 2004, Issue 16

Loss of complexity of trabecular bone with aging

Loss of complexity of trabecular bone with aging

Iliac crest bxIliac crest bx femoral headfemoral head lumbar spinelumbar spinelumbar spine

37 year old37 year old

84 year old84 year old

Pictures from Ralph Muller

Ropp

Fopp

C

Remodelling = accumulation of Robustness (R)+

accumulation of Frailty (F)+

loss of Complexity (C)

individualvariability

(geneticallydetermined)

0 10070AGE

accumulation of Robustness (R)+

accumulation of Frailty (F)+

loss of Complexity (C)

increased role ofstocasticity

with age

Some characteristics of the centenarians

CLINICAL CHARACTERISTICS OF THE STUDY GROUPS (n=466)CLINICAL CHARACTERISTICS OF THE STUDY GROUPS (n=466)

CONTROLS n = 298

CENTENARIANSn = 168

Sex (male/female)

Plasma triglycerides (mmol/l) 1.5 ± 0.8 1.2 ± 0.4*

Plasma total cholesterol (mmol/l) 5.5 ± 1.1 4.5 ± 1.1*

Plasma HDL cholesterol (mmol/l) 1.4 ± 0.4 1.6 ± 0.4*

Plasma LDL cholesterol (mmol/l) 3.2 ± 1.1 1.6 ± 0.8*

IR (HOMA) 3.9 ± 1.7 1.5 ± 0.7*

26.1BMI (Kg/m2)

6.0

14.8

2.4± 22.3 ± 3.1*Plasma Glucose (mmolL) ± 4.1 ± 1.1*

Plasma Insulin (mU/l) ±

1.2

6.9 ± 2.8*6.4

148/150 51/117

*p<0.01 vs control group

Paolisso et al., Exp. Gerontol., 37, 149-156, 2001

Age (years)

IR (H

OM

A)

1

2

3

4

5

6

20-40 40-50 50-60 60-70 70-80 80-90 90-100 >100

p for trend < 0.01

INSULIN RESISTANCE IN THE WHOLE POPULATION (N = 466)INSULIN RESISTANCE IN THE WHOLE POPULATION (N = 466)

Paolisso et al., Exp. Gerontol., 37:149-156,2001

Peculiar energy metabolism andIGF-1/Ins signaling in

centenarians

• Is the lack of insulin resistance a pre-requisite for longevity?

• Do centenarians perform better as far asglucose metabolism and energymetabolism is concerned?

Centenarians remindcalorically-restrictedhormetically-adapted

organisms

IGF-1 plasma levels are very low in centenarians

Bonafè et al., 2003

IGF1 plasma levels are correlated withpolymorphisms of IGF1R and PIK3 kinase

Trade off between muscle strengthand cancer in aged people

- High plasma levels of IGF-1 (genetically controlled)) are beneficialin aged people, especially regarding physical performance and the maintenance of muscle strength

- The price to pay is likely an increased risk of cancer

- Low plasma levels of IGF-1 (genetically controlled) are beneficial in the last decades of life (nonagenarians and centenarians) and are associated with lower propensity to cancer growth

- The price to pay is frailty and massive reduction of musclestrength

- High levels of IL-6 are always detrimental for muscle mass and strength as well as for late survival

Proteasomeand

Immunoproteasome

Proteasome activity analysis in cultures of embryonic cells (MRC5; passage 21), controldonors (28 and 80 years old; passage 9) and centenarians C1, C2 and C3 (passage 9; greybars), as well as in C6 culture undergoing replicative senescence (black bars).

Chondrogianni et al. Exp Gerontol., 2000

Proteasome activity is maintained in fibroblasts fromcentenarians and decreases with in vitro passages

RNA expression level of proteasomesubunits is maintained

Chondrogianni et al. Exp Gerontol., 2000

Evolution, Ageingand

Inflammation(Inflamm-aging)

Evolutionary Medicine

1. our body has been selected for (millions of years) in order to survive until the age of reproduction

2. the average life expectancy at birth was quite short (about 40 years) until a couple of centuries ago

3. our defence and repair systems have been moulded by evolution to

cope with all sort of external (pathogens) and internal (ROS)

stressors and

to maintain fully efficient bodies for and until reproduction

4. our immune system has also been moulded by evolution and it is quite efficient in coping with acuteinfections in young people;

5. the same system is not particularly efficient in responding to chronic stimuli in old subjects;

6. both innate immunity and clonotypical(adaptive) immunity appear to be

hyper-stimulated in old peopleowing to

decades of evolutionary unpredicted antigenic stimulation, mostly due to

sub-chronic, bacterial and viral infections

we are “immunologically unfit”

The new paradigm of evolutionary medicine

the most important pathologies involvegenes and gene variants (polymorphisms) which have been selected to adapt to anenvironment characterized by :

- Famine ( today plenty of food)- Infections (today hygenized

environment, food…)- Great expenditure of calories (today

heating, low physical excercise)

Pathological conditionswith an inflammatory

pathogenesis(or an inflammatory strong component)

• 1. atherosclerosis, MI, stroke• 2. metabolic syndrome, obesity, type 2

diabetes• 3. osteoporosis and osteoarthritis• 4. neurodegeneration ( Alzheimer’s disease)• 5. cancer• 6. major depression• 7. FRAILTY

Ann. N.Y. Acad. Sci., 908, 244-254, 2000

repeated episodes of acute or chronic stressphysical or psychological

Cytokines Chronic InflammationHormones (Inflamm-ageing)

repeated episodes of acute or chronic stressantigenic

Franceschi et al., Inflamm-aging: an evolutionary perspective on Immunosenescence

Ann. N. Y. Acad. Sci. 908, 879-96, 2000

Antigenic stimulation and pro-inflammatory cytokines increase with age

De Martinis et al., 2005

innate immunity, stress response and inflammationare evolutionary interconnected

and macrophage-centered

Franceschi et al., 2000

Obese adipose tissue is charactherized by inflammation and progressive infiltration by macrophages as obesity develops

Immuno-activation, Inflammation and Obesity

The filling up of the immunological spacein old subjects

EffectorsMemory

OldVery young Young CD8+

T Cells

Naive

Chronic cytomegalovirus (CMV) infection plays a major role

in immunosenescence

The immune system of old peopleis full of megaclones specific

for CMV epitopes

Virus Viral protein Epitope Sequence HLA-restriction

Tetramers

EBV BMLF1 lytic protein

280-288 GLCTLVAML A*0201 EBV1

EBV EBNA3A Latent protein

379-387 RPPIFIRRL B*0702 EBV2

CMV pp65 495-503 NLVPMVATV A*0201 CMV1

CMV pp65 417-426 TPRVTGGGAM B*0702 CMV2

MHC- class I-restricted viral epitopes and related tetramers used to assess the number of circulating CMV-

specific CD8+ T cells

Sansoni et al., Exp.Gerontol., 39, 1233-43, 2004

Donor Age HLA %EBV1+/CD8+(n°/µL)

%CMV1+/CD8+(n°/µL)

%EBV2+/CD8+(n°/µL)

%CMV2+/CD8+(n°/µL)

001 97 A2+ <0.01 (0) 1.7 (12)

002 93 A2+ 0.58 (1.3) 13.4 (30.8)

003 91 A2+ <0.01 (0) 0.6 (1.4)

004 102 A2+ <0.01 (0) 4.9 (25)

005 99 A2+ 0.9 (2.3) 0.54 (1.4)

006 95 A2+ <0.01 (0) 1.5 (2)

012 101 A2+/B7+ <0.01 (0) <0.01 (0) 0.22 (0.9) 1.03 (4)

013 99 A2+/B7+ <0.01 (0) 0.19 (1.1) 0.06 (0.3) 2.8 (17)

014 102 A2+/B7+ <0.01 (0) <0.01 (0) 0.2 (1.8) 22.2 (208)

015 102 A2+/B7+ <0.01 (0) 0.3 (1) 0.77 (2.6) 5.9 (20.2)

016 99 A2+/B7+ <0.01 (0) <0.01 (0) 0.3 (1) 2.27 (8)

017 94 B7+ 0.12 (0.4) 5.5 (18)

018 93 B7+ 0.5 (1.7) 1.6 (5.6)

019 93 B7+ 0.2 (0.6) 1.2 (3.6)

020 104 B7+ <0.01 (0) 0.6 (5)

JAGS, 53, 906-8, 2005

CMV infection and Frailty in the Elderly

The combination of CMV infection + high plasma levels of IL-6

synergizeregarding frailty in the elderly

Schmaltz et al., 2005

THE GUT: an important source of antigenic stimulation in elderly and frail people

Memory cellsare

stimulated and expandedby cross-reactive antigens

The filling of the immunological space

Franceschi and Bonafè, 2003

The absolute number of virgin (non antigen-experienced)T cells (0) CD95- CD28+) decreases linearly with age

and there is an exaustion of such cells in centenarianswhich is correlated to an increased risk of mortality

(Fagnoni et al., Blood, 2000)

Thymus from a 5 years old boy Courtesy of M. Capri

Thymus from a 54 years old manCourtesy of M. Capri

Thymus from a 74 years old manCourtesy of M. Capri

Thymic output with age

n.16 n.13 n.25

Nasi et al., Aging Cell 2006 in press

Plasma levels of IL-7 do not change with ageand are higher in female than in male centenarians

Immune status in very elderly IRP and non-IRP donors

Alterations with agenon-IRP IRP

Marker and cells

CD4:CD8 > 1 CD4:CD8 < 1

T cell proliferation→→

T cell proliferation ↓

CD28 ↑ CD28 ↓

CD57 ↓ CD57 ↑

CD45RA ↑ CD45RA ↓

CD45RO ↓ CD45RO ↑

KLR-G1 ↓ KLR-G1

Cytokines and growth factors

IL-2 ↑ IL-2 ↓

IL10 → IL-10 →

IFN-g ↑ IFN-g ↓

CMV/EBV status

CMV+/ lower frequencies

Mostly KLR-G1+

CMV+/ higher frequencies

Mostly KLR-G1+

EBV+/ lower frequencies

EBV+/ higher frequencies

The individual threshold for DDD

Franceschi and Bonafè, Biochem. Soc. Transact. (2003) vol. 31, part 2

Context and Noise

the problem of context in aging research

aged cells in aged bodies:

intrinsic (?) extrinsic (?) (cell) (environment)

which is the more important ?

repeated episodes of acute or chronicpsychological and physical stress

Cytokines Chronic InflammationHormones Inflamm-aging

repeated episodes of acute or chronicantigenic stress

drastic change ofmicroenvironment

the role of the CONTEXTin the aging process

immunological / inflammatory“noise / field” can be

pathogenic(abundance, pleiotropy and redundance of

cytokines and growth factorsboth sistemically and locally)

Stem cells(hemopoietic progenitors)

Hemopoiesis in centenarians

- the number of CD34+ hemopoietic progenitor cells(HPC) in the peripheral blood of centenarians isabout 50% of that found in young subjects

- when put in optimal culture conditions CD34+ cellsfrom centenarians are perfectly able to respond togrowth factors and form erythroid, granulocyte/macrophage and mixed megakaryocyticcolonies in a way (number, size and morphology) indistinguishable from that of young subjects

- a remodelling of the production of hemopoieticgrowth factors occurs in old people and centenarians

IL-3

GM-CSF

SCF

What changes dramatically with age isthe number of HPC and their

environmentbut not their functional capability to

differentiate

“old” satellite cells are perfectly ableto generate new muscle fibers whenexposed to a “young” environment

and viceversa that“young” satellite cells are inhibited by

the old milieu

Nature 433, 760-764, February 2005

abnormal and aberrantmitotic stimuli

in the microenvironment(inflamm-aging ?)

Narita and Lowe, Nature Med, 2005