THE INFLAMM-AGING THEORY

An Evolutionary Perspective onImmunosenescence and Aging

Dr. J. Alijotas-Reig*¶ ♦, R. Kandhaya-Pillai¶.

*Systemic Autoimmune Diseases Unit. Service of Internal Medicine-I. Vall d’ Hebron University Hospital. UAB (Prof. M. Vilardell-Tarrés)

¶ Aging Basic Research Unit. Molecular Biology and Biochemistry Research Center for Nanomedicine. CIBBIM-Nanomedicine. Vall d'Hebron University Hospital. Research Institut (VHIR)

♦ Department of Medicine. Faculty of Medicine. Universitat Autònoma de Barcelona. Spain

2012-UPDATED

Maybe, in that moment, he loved the life more than ever…… What he desired were the same answers we all look for.

• Where do I come from?

• Where do I go?

• How long am I going to stay?

Blade Runner. Ridley Scott, 1982

For human being, aging have more questions than answers.

TEORIAS MOLECULARES Y GENÉTICASMutaciones somáticas y alteraciones en la reparación del ADNErrores catastróficos de transcripción o de translocaciónAlteraciones o diversidad de gerontogenes (LAG-1)Telómeros / Longuitud telomerasasPresencia de apoE4Mutación del precursor de la beta amiloide

TEORÍAS CELULAREST. de HayflickT. de los radicales libres (R.O.S)T. mitocondrialT. de la glicosilación proteicaT. de las membranas

TEORÍAS DE SISTEMAST. inflamatoria, (INFLAMMAGING THEORY)

T. Neuroendocrina

PRINCIPALES TEORIAS SOBRE EL ENVEJECIMIENTO

Could aging be a chronic low-grade inflammatory state?

InflammationSenescence

Cancer

Autoimmunity

INFLAMMAGING THEORYChronic low-grade inflammation

Pro-inflammatory cytokines Frailty

Atherosclerosis

Parkinson disease

Alzheimer disease

Osteoarthritis

Osteoporosis

Insulin resistance/type 2 diabetes mellitus

Sarcopenia

Predisposed genetic background Predisposed genetic backgroundMacrophage

INFLAMMAGING THEORY

Coined by Claudio Franceschi * in 2000

Ageing is accompained by a low-grade, systemic up-regulation of the inflammatoryresponse.

Chronic low-grade inflammatory status

Senescence age-associated diseases

Franceschi et al, Ann NY Acad Sci, 2000

OsteoporosisOsteoarthritisSarcopeniaAlzheimerDiabetes MellitusArtheriosclerosisParkinson disease

INFLAMMAGING THEORY

This inflammatory state is characterised by:

1. Low grade

2. Controlled

3. Asymptomatic

4. Chronic

5. systemic

INFLAMMAGING THEORY

Internal and environmental inflammatory stimuli

individual background

Pro-inflammatory cytokines

cytokine polymorphisms

Reactive Oxigen Species (ROS) oxidative damage

Autoantibodies

pro-inflammatory cytokines

INFLAMMAGING, CYTOKINES AND AGING

The most important age-associated diseases share a common inflammatory background:

TNF-α, TNF-β, IL-1β, IL-6, IL-10 are the most proinflammatory involved cytokines

Certain cytokine polymorphisms have been related to aging:IL-10 (1082 G/A)TNF-αIL-6 (174 G/C)

Aging: gender differences

Role of Toll-like receptor 4 (TLR4) and Heat shock proteins (HSPs)

INFLAMMAGING, CYTOKINES AND AGING

• Inflammation, metabolism and aging:

muscle mass

adipose tissue

mitochondrial function

TNF-αLeptinaIL-1IL-6IFN-γIL-8FibrinogenAngiotensinogenCRPPAI-IMetallothioneinComplement factor 3Monocyte chemotactic protein-IMacrophage inflammatory protein-ISAA protein

B.M.I

Cell energy R.O.S productionCell apoptosis

DNA maintenance DNA damage

IL-6 / TNF-α / IL-1

AGEING

SISTEMA INMUNE Y ENVEJECIMIENTO

Alteraciones en la producción de citocinas:

predominio de Th1 sobre Th2

IL-2 ? / IL-2R IL-3?

IL-6 IL-4

α-TNF IL-5

IL-10

APCs IL-1

nTreg ? pTreg?

Immunological polymorphismsprofiles and ageing

• ↑ IL-2• ↑ IL-7 +Zn• ↑ IL-4• ↑ IL-10• ↓ TNFα• ↑ CD45RA+ y ↓ CD45RO+• ↓ IL-6• ↓ IFN-gamma• ↑nTreg (CD4+CD25-Foxp3+)

↓ Th1 / ↑ Th2 / ↑ nTreg

Down-regulation

IMMUNOSENESCENCE

Thymosuppressive Thymostimulatorycytokines cytokines

steroids LIF IL-7IL-6 KGF

Oncostatin MTSLP

hGHLeptinIL-10

ThymusRobustThymusAtrophied

thymus

IL-1, IL-3, TGF-β

DNA-anti-apoptotic Bcl-2

AGEING: IS IT AN AUTO-[INNATE]-IMMUNITY SUBCLINICAL SYNDROME?

Classical inflammatory signs

Redness (rubor)

Swelling (tumor)

Pain (dolor)

Warm (calor)

Loss of function (functio lesa)

Inflammaging cardinal signs

Low-grade

Controlled

Asymptomatic

Chronic

Systemic

Giunta S. Immunity&Ageing, 2007

May be inflammaging a new chapter of autoimmunity?

INFLAMMAGEING THEORY

Clasical autoimmune diseases adaptive immunity

New “innate” autoimmune syndrome innate immunity

“Inflammaging may constitute the subclinical paradigm of new type of autoimmunity, namely that arising from an auto-immune inflammatory response of the innateimmune-syndrome an old actor of immunity and yet a new actor of autoimmunity”*

*Giunta S. Immunity&Ageing 2006

CELLULAR SENESCENCE CAN BE INDUCED BY

SENESCENCE CELLS

MORPHOLOGICAL CHANGESCells become largerCells display increasedgranularityCells flatten outCell borders tend to vanish

SENESCENCE CELLS

BIOCHEMICAL CHANGES(beta)galactosidase activity +

Questions to be answered:

In an in-vitro cellular model, may low-level proinflammatory cytokines provoke premature cell senescence?

Which are the intracellular pathways used, through which proinflammatory cytokines provoke their undesirable cell actions regarding cellular senescence?

Which is/are the gene/s that regulate/s the final expression of these proinflammatorybackground or what gene/s may act as a protective against the former proinflammatory status? (robust or frail gene ?)

Molecular Regulation of Inflammatory Pathways in Human Endothelial Cells Senescence

To establish a cellular model of senescence by inflammatory signals

To study the gene expressional changes and signaling pathwaysregulated during senescence.

Dissect the molecular role of inflammatory pathways during senescence

To determine the miRNA architecture in senescence

Molecular Regulation of Inflammatory Pathways in Human Endothelial Cell Senescence

Resultados

Tto crónico con TNFα induce senescencia prematura en células HUVEC

↑ROS en HUVEC en respuesta a estimulo crónico con TNFα

Diferencias en la expresión de micromatrices de RNA

Diferencias en la expresión de miRNA durante la senescencia inducida por TNFα

β-GalP53, p21BrdU /IPH2AX

H2AX

Sobre expresión de 341 genesExpresión suprimida de 40 genes(transcritos en 5 clústeres)

Importante: circuito TNFα IL-6 STAT1/STAT3 (via p38/MAPK)

Molecular Regulation of Inflammatory Pathways in HumanEndothelial Cell Senescence

Diferencias en la expresión de miRNA durante la senescencia inducida por TNFα

HUVEC “jovenes”/HUVEC senescentes (rep)

↑ miR 217↑ miR 145

↓ miR18a↓ miR21

HUVEC-TNFα/HUVEC sen replicativa

↑ miR 146a↑ miR 21↑ miR 195

↓ miR 7c

Dianas potenciales: KIF2C, SPC25, CDC25BNCAPG, RAD51AP1,HMGA2, KIF2C, CEP55/G0S2

Resultados:

Molecular Regulation of Inflammatory Pathways in HumanEndothelial Cell Senescence

La estimulación crónica con TNFα se relaciona con la senescencia prematura.

La activación STAT1/3-IL6 promueve un asa amplificadora autoregulada que perpetua la vía inflamatoria.

Además de STAT1/3, la via JAK2 también juega un rol importante (doble via JAk2/STAT1/3).

La senescencia inducida por TNF-α se acompaña de modificaciones en la expresión de diversos genes responsables del control del ciclo celular.

Determinadas alteraciones a nivel del miRNA sugiere su posible papel en la regulación de la senescencia.

CONCLUSIONES:

Inflammation, autoimmunity and ageingAPOPTOSIS, mediated by two major pathways:

Extrinsic Intrinsic

death receptor-mediated (TNFRs) mitchondria mediated

(CD95)

trimerization of DD poor studied in elderly

pro-caspases

caspases

Inflammation, autoimmunity and ageing

APOPTOSIS IN AGINGIncreased apoptosis in T cells, CD4+ and CD8+

↑ sensitivity CD95-CD95L ↑ activation caspase-8 /3

↑ FADD

↑sensitivity to TNF-œ -induced apoptosis

(to analize the proposed mechanisms, see figure 3)

Inflammation, autoimmunity and ageingRole of apoptotic cells in the regulation of inflammation and

changes in aging (I)

Apoptotic cell death and clearance of dead cells is essential to homeostasis

Clearance of apoptotic cells have an anti-inflammatory effects

Ie: co-cocultures between apoptotic cells and APCs

Apoptosis regulates the cytokine profile

Slow apoptotic cell clearence undergo secondary necrosis

Dendritic cells (APCs) are basic in large scale phagocytosis apoptotic cells

Inflammation, autoimmunity and ageingRole of apoptotic cells in the regulation of inflammation and

changes in aging (II)

Apoptotic cell death and clearance of dead cells is decresed in elderly

Phagocytosis of apoptotic T-cells by dendritic cells is decreased

TNF and IL-6 of apoptotic cells is increased (old/young)

Co-cultures failed to inhibid the production of pro-inflammatory cytokines

Accumulation of apoptotic cells, inducing secondary necrosis and inflammation

DCs may uptake self-Ag derived of apoptotic cells or 2 necrosis to induce

autoimmunity.

Failure in apoptotic regulation: INFLAMMATION AND

MECHANISMS OF IMMUNOSENESCENCE

Three major models of human accelerate immune aging

• Latent CMV infection• HIV infection under highly active antiretroviral therapy (HAART)• Chronic autoimmune disease

Chronic or latent viral infections CMV / HIV

Lymphocyte exhaustionAccelerate immune aging

MECHANISMS OF IMMUNOSENESCENCEThymic involution accelerates immune aging