cours M2

Le Stress inflammatoire

Francis Berenbaum Universit茅 Pierre & Marie Curie AP-HP, h么pital Saint-Antoine, Paris

OSTEOARTHRITIS

NORMAL

OSTEOARTHRITIS Subchondral cyst Cartilage

Capsule

Synovial membrane

Subchondral osteosclerosis

Cartilage loss

Synovitis Osteophyte Cartilage erosion

Thickening of the capsule Os sous-chondral

Bijlsma, Berenbaum, Lafeber. The Lancet (2011)

OA : a disease of the whole joint

Sellam & Berenbaum. Nature Reviews Rheum Nov. 2010

OA : the synovial perspective

(personnal collection)

PATHOPHYSIOLOGY OF OSTEOARTHRITIS 20th century

21st century

•A passive consequence of tear and wear •A cartilage disease •A degenerative disease

•A cell-driven mechanism based on soluble mediators •A multiple-tissue disease •A pro-inflammatory disease

Stress inflammatoire et cartilage

1- Chondrocytes are involved in the cartilage degradation process

QUANTITATIVE MODIFICATIONS •matrix degradation +++ •Apoptosis of chondrocytes

NORMAL

OA

QUALITATIVE MODIFICATIONS •Synthesis of a low-quality matrix •Modification of the differentiation state of chondrocytes •Matrix calcification

PATHOPHYSIOLOGY OF OSTEOARTHRITIS

Anabolism Cartilage matrix synthesis

Catabolism Cartilage matrix degradation

Homeostasis

PATHOPHYSIOLOGY OF OSTEOARTHRITIS

Anabolism

Catabolism

Cartilage matrix synthesis dise qu

Normal

ilibrium

Overexpression of Metalloproteinases 1,3,13 by chondrocytes Osteoarthritis

LES METALLOPROTEASES

PATHOLOGY OF OSTEOARTHRITIS

Anabolism Cartilage matrix synthesis dise qu

Normal

Catabolism Soluble mediators : IL-1,PGE2‌ ilibrium

Overexpression of Metalloproteinases 1,3,13 by chondrocytes Osteoarthritis

OA : the cartilage

cartilage degradation

Synthesis of an altered matrix

Proteolysis

Proteinases (MMP, ADAMTS)

Les médiateurs impliqués • • • • •

Métalloprotéases Cytokines et facteurs de croissance Dérivés oxygénés prostaglandines Fragments de matrice

ACTIVATION

Noyau

COX-2

chondrocyte MMPs MĂŠdiateurs inflammatoires

Médiateurs prodégradatifs •IL-1 •TNFα •IL-17 •prostaglandines •ATP •…

Voies de signalisation inductibles

régulation transcriptionelle et post-transcriptionelle COX-2

MMPs Médiateurs inflammatoires

médiateurs pro-dégradatifs

NF-kB, MAPK, Wnt, autres

•IL-1 •TNFα •IL-17 •PGE2 •ATP •…

régulation transcriptionelle et post-transcriptionelle COX-2

MMPs Médiateurs inflammatoires

VOIES DE SIGNALISATION DU CHONDROCYTE

Goldring Best practice, 2006

Interaction Ligands/specific receptors

•Cytokines (including chemokines, adipokines) •Prostaglandins •Reactive oxygen species •RAGE ligands •Extracellular matrix components

MAPK NF-kB Others…

Transcriptional and post-transcriptional regulation COX-2

MMPs ADAMTS Cytokines Reactive oxygen species Prostaglandins

Vieillissement du cartilage et stress inflammatoire

Formation d’AGE dans le cartilage Autofluoresence

Caramélisation du cartilage

68 years

AGE fluorescence/Hyp RFU/nmol

14 years 30000 R2 = 0.5539

20000

Knee Ankle

54 years 2

10000

R = 0.5537

0 0

20

40

60

Age in Years

80

100 Knee p = 0.02 Ankle p = 0.03

Loeser et al.

« AGE » ET ARTHROSE

« AGE » (« Advanced Glycation End-products ») :

Liaison covalente sucre-AA

Versijl et al. , Current Opin Rheumatol 2003

OA and inflammation : role of aging IL-1 stim./Control MMP-13

8 6

r=0.57 p=0.0001

4 2 0 0

10 20 30 40 50 60 70 80 90 Donor Age

Chondrocytes were treated overnight with 5ng/ml IL-1β and MMP-13 was measured by immunoblotting. Similar to fibronectin fragments, IL1β stimulated MMP-13 increased with age.

Forsyth et al, J Gerontol Biol Sci, 2005

Stress inflammatoire et tissu synovial

THE SYNOVIAL TISSUE IS INVOLVED IN THE OA PROCESS

Normal Synovial tissue

Fibrosis

Mild Inflammation

severe inflammation

TISSU SYNOVIAL ARTHROSIQUE

Immunostaining of Chemokines and MMP in OA Synovium Control

MCP-1

MMP-1

MMP-3

Synovial Mononuclear Cell Infiltrates in OA T-Cells (CD3)

B-Cells (CD20)

Plasma Cells (CD138)

Da et al., J Immunol., 2007 Sakkas et al., Arthritis Rheum

Cells of lymphoid origin may contribute to the pathophysiology of OA

Depletion of Synovial Macrophages Reduces Cartilage Damage DIPEN Staining (black)

•

Murine Collagenase-Induced OA

•

Clodronate (liposomal) utilized to deplete synovial macrophages Control

•

Cartilage damage determined by histopathology and DIPEN immunostaining (Neo-epitope for MMP activity)

Blom et al. Arthritis Rheum. 2007

Macrophage Depleted

Synovial macrophages play a role in OA cartilage pathology

Stress inflammatoire et os sous chondral

BONE AND OA

Bone-Cartilage Unit in OA Normal

OA Erosion

Articular cartilage

Tidemark

Calcification

The Bone-Cartilage Unit

From Lories & Luyten, Nature Rev Rheum 2011

Cartilage and Bone in OA

- Permeability of calcified cartilage to small molecules (~ 400 Da) Arkill, Osteoarthritis Cartilage 2008

Cartilage

- Diffusion of fluorescein sodium (360 Da) from subchondral bone to cartilage

Calcified cartilage

Subchondral bone

Pan, J Orthop Res 2009

The Bone-Cartilage Unit

Lories & Luyten, Nature Rev Rheum 2011

A new model for assessment of bone-cartilage communication Mechanical stress applied Mechanical stress appliedon on Flexercell Compression Plus FX-4000C osteoblast-derived matrix osteoblast-derived matrix::

Biopress

10% = 1,67 MPa 1 Hertz

medium

compression

Articular chondrocytes

Bone-cartilage communication

Osteoblast-derived medium induces MMP-13 protein expression in chondrocytes

MMP-13 secretion (Western Blot quantification) UC =uncompressed C =compressed UC = medium from uncompressed osteoblasts CM = medium from compressed osteoblasts

UC

C

Osteoblasts

Control UCM

CM

Chondrocytes

Priam et al. (in preparation)

« Genes and Osteoarthritis »

•Musculoskeletal ageing

•Matrix frailty

•Diabetes

•MetS •Hormones Systemic factors

•Post-trauma •Dysplasia Obesity Local factors

Obésité et stress inflammatoire

THE CONCEPT OF ADIPOCYTOKINES (OR ADIPOKINES)

Tilg & Moschen, Nature Reviews Immunology, 2006

PROTEINS SECRETED BY ADIPOSE TISSUE •Adiponectin •Resistin •Leptin •TNFα •IL-6 •Vaspin •Omentin •Adrenomedulin •Retinol Binding Protein-4

•Angiotensinogen •Adipsin •Fasting-induced adipose factor •PAI-I •Monocyte chemoattractant protein-I •TGF-β •Apelin •Acylation stimulating protein •Visfatin/PBEF/NAMPT

THE PREVALENCE OF HAND OA IS INCREASED IN OBESE PATIENTS !

~TWO-FOLD MORE RISK TO HAVE HAND OA IN OBESE POPULATION !

Yusuf et al. Ann Rheum Dis 2009 (epub)

Systemic factors

Mechanical stress

ROLES OF ADIPOKINES IN ARTHRITIS

From Lago et al. NCPR 2007

A NOVEL PARADIGM FOR THE PATHOPHYSIOLOGY OF OA

Trauma Local inflammation

MetS Low-grade inflammation

OSTEOARTHRITIS

ageing Secretory Inflammatory phenotype

OA induces experimental Alzheimer’s disease

APP/PS1;Col1-IL1βXAT model Kyrkanides et al. J Neuroinfl 2011

A NOVEL PARADIGM FOR THE PATHOPHYSIOLOGY OF OA

Trauma Local inflammation

MetS Low-grade inflammation

ageing Secretory Inflammatory phenotype

OSTEOARTHRITIS

Systemic release of inflammatory mediators

Alzheimer

Atherosclerosis ? Others ?

Pierre & Marie Curie University Paris VI « Joint ageing, cartilage degradation and inflammation» (F. Berenbaum) • • • • • • • •

Xavier Claire Marie-Charlotte Zvezdana Geoffroy Audrey Sabrina Jeremie

Houard (MCU) Jacques (MCU) Laguillon (Thésarde) Mladenovic (Tec) Nourissat (Thésard) Pigenet (tec) Priam (Thésarde) Sellam (MCU-PH)

Adipokines : VISFATIN+++

OSTEOARTHRITIS : A better understanding of the disease for a better treatment Prof. Francis BERENBAUM Pierre & Marie Curie University AP-HP Saint-Antoine hospital, Paris, France

Stockholm, Oct 19th 2011

OA : the bone perspective Spontaneous OA : Increase in subchondral bone cartilage destruction (guinea pig)

density

BEFORE

Experimental post-trauma induced OA : alendronate (inhibiting bone remodeling) decrease cartilage degradation (mice)

bone cells-chondrocyte co-culture : Increase in biomarkers of cartilage degradation (humans)

Bone-Cartilage Unit in OA Normal

OA Erosion

Articular cartilage

Tidemark

Calcification

The Bone-Cartilage Unit

From Lories & Luyten, Nature Rev Rheum 2011

The Bone-Cartilage Unit

Lories & Luyten, Nature Rev Rheum 2011

A new model for assessment of bone-cartilage communication Mechanical stress applied Mechanical stress appliedon on Flexercell Compression Plus FX-4000C osteoblast-derived matrix osteoblast-derived matrix::

Biopress

10% = 1,67 MPa 1 Hertz

medium

compression

Articular chondrocytes

Bone-cartilage communication

Osteoblast-derived medium induces MMP-13 protein expression in chondrocytes

MMP-13 secretion (Western Blot quantification) UC =uncompressed C =compressed UC = medium from uncompressed osteoblasts CM = medium from compressed osteoblasts

UC

C

Osteoblasts

Control UCM

CM

Chondrocytes

Priam et al. (in preparation)

PATHOPHYSIOLOGY OF OSTEOARTHRITIS : background for targeted therapies

TARGET !

adipokines

TARGET !

TARGET !

A targeted DMOAD : The Quest for the Holy Grail ?

-

MMP inhibitor : negative results (and side-effects) Risedronate : negative results Oral calcitonin : negative results IL-1RA : negative results Anti-TNF : negative results rhFGF18 : ongoing (presented at OARSI 2011) Strontium ranelate : ongoing

CLASSIFICATION OF ARTHRITIS

CLASSIFICATION OF ARTHRITIS

Risk factors for incidence and progression of OA of the knees, hips and hands

Bijlsma & Klahr. Best Pract Res Clin Rheum 2007

« Mechanical-induced Osteoarthritis »

•Post-trauma •Dysplasia •Overweight Local factors

Role of mechanical stress Mechanical stress

Interaction Ligands/specific receptors

•Cytokines (including chemokines, adipokines) •Prostaglandins •Reactive oxygen species •RAGE ligands •Extracellular matrix components

MAPK NF-kB Others…

Transcriptional and post-transcriptional regulation COX-2

F. Berenbaum Primer in the Rheumatic Diseases, 13th Ed. (dec 07)

MMPs ADAMTS Cytokines Reactive oxygen species Prostaglandins

MECHANORECEPTORS ON CHONDROCYTES

COX-2

mPGES-1

Gabay et al. Osteoarthritis Cartilage 2008 Gosset et al. AR&T 2006

’ 60 d Lo a

de

d de Lo a

Un lo

ad

ed

30

’

Mechanosignalling and OA

***

**

Obesity, Trauma

Mechanical stress Mechanical stress

chondrocyte

COX-2

Loss o f

cartila

ge hom

eostas

is

Osteoarthritis

•

A 2-fold increase of hand OA in obese patients !

RR = 1,9

Yusuf et al. Ann Rheum Dis 2009

Systemic factors

Mechanical stress

« Metabolic-induced Osteoarthritis »

Obesity

Systemic factors

•Post-trauma •Dysplasia

Local factors

OA and adipokines

Gabay & Berenbaum. Curr Rheum Reviews 2009

Obesity

Mechanical stress

Adipokines

Mechanical stress

chondrocyte

COX-2

Loss of cartila ge

homeo

stasis

Obesity-induced OA

• •

Cohort of 482 women (47 years-old) Prevalence of knee OA = 11%

Obesity

« cardiometabolic status » (≥ 2 criteria)

BMI >30

•History of diabetes, antidiabetic drugs, glycemia > 126 g/dL •CRPus ≥ 2 mg/L •HDL ≤45 mg/dL or LDL>160 mg/dL •TG ≥ 200 mg/dL •height/Hip ≥ 0.81 cm •Systolic BP > 135 mmHg, diastolic BP > 85 mmHg or anti-hypertensive drugs Sowers et al. A Rheum Oct 2009

Risk for OA according to BMI and cardiometabolic clustering With knee OA n(%)

Odds ratio (IC 95%), p

Non-obese with NO cardiometabolic parameters (n=212)

10 (4,7%)

1

Non-obese WITH cardiometabolic parameters (n=85)

5 (5,9%)

1,28 (0.43-3.87)

Obese with NO cardiometabolic parameters« (n=47)

6 (12,8%)

3.00 (1.03-8.71), p<0.05

Obese WITH cardiometabolic parameters« (n=138)

32 (23,2%)

6.2 (1.93-13.07), p<0.0001

In obese patients, knee OA more frequent when cardiometabolic parameters are present Sowers et al. A Rheum Oct 2009

« Metabolic-induced Osteoarthritis »

•MetS

Obesity

Systemic factors

•Post-trauma •Dysplasia

Local factors

OA is associated with MetS NHANES III Data â&#x20AC;˘Comparaison of the prevalence of the MetS in subjects w/o OA â&#x20AC;˘To determine whether having OA predicts increased CV risk 59% 23%

44 years old man with OA : Risk of having MetS

x5

!

Puenpatom et al. Postgrad Med J 2009

Each of the 5 CV risk factors are more prevalent in the OA population NHANES III Data â&#x20AC;&#x201C; General population

Puenpatom et al. Postgrad Med J 2009

Ann Rheum Dis 2011

« Metabolic-induced Osteoarthritis »

•MetS •Diabetes

Obesity

Systemic factors

•Post-trauma •Dysplasia

Local factors

OA and atherosclerosis • In women, association between hand OA and : – Coronary calcifications OR = 1,42 (1,14-1,76) p=0,002 – Carotids plaques OR = 1,25 (1,04-1,49) p=0,016 • Adjusted on age, smoking, cholesterol, triglycerides, BMI, BP, statins

• Association between popliteal arterial wall thickness and generalized OA (adjusted on sex, age, BMI) Jonsson ARD 2009, GARP Jonsson ARD 2009, AGES Reykajvik Study

Similarities between OA and atherosclerosis â&#x20AC;˘ -

â&#x20AC;˘

Osteoarthritis

Age Obesity Failure of repair damage cartlage Destruction by proteinases Aneural articular Integrated role of cartilage and bone Synovial macrophages produce inflammatory mediators

-

Atherosclerosis

Age Obesity Failure of repair damage endothelium Destruction by proteinases Aneural atherosclerotic matrix Integrated role of endothelium and SM cells macrophages produce inflammatory mediators

2011

• • •

n=1163 (southwest England) , >35 years old with knee or hip symptomatic Mean follow-up : 14 years Comparison with general population, same age and gender (SMR:standardized mortality ratios)

Excess mortality in OA patients, mainly due to CV events.

Gabriel and Michaud Arthritis Research & Therapy 2009

« Aging-induced Osteoarthritis »

•Musculoskeletal ageing

•Matrix frailty

•Diabetes

•MetS •Hormones Systemic factors

•Post-trauma •Dysplasia Obesity Local factors

AGE Formation in Cartilage Autofluoresence

Cartilage caramelisation

68 years

AGE fluorescence/Hyp RFU/nmol

14 years 30000 R2 = 0.5539

20000

Knee Ankle

54 years 2

10000

R = 0.5537

0 0

20

40

60

Age in Years

80

100 Knee p = 0.02 Ankle p = 0.03

Loeser et al.

« AGE » AND OA

« AGE » (« Advanced Glycation End-products ») :

covalent binding between sugar and AA

Versijl et al. , Current Opin Rheumatol 2003

Aging increases the response to IL-1β IL-1 stim./Control MMP-13

8 6

r=0.57 p=0.0001

4 2 0 0

10 20 30 40 50 60 70 80 90 Donor Age

Chondrocytes were treated overnight with 5ng/ml IL-1b and MMP-13 was measured by immunoblotting. Similar to fibronectin fragments, IL-1b stimulated MMP-13 increased with age.

Forsyth et al, J Gerontol Biol Sci, 2005

Senescence-associated secretory phenotype

CoppĂŠ et al. Annu Rev Pathol 2010

Senescence-associated secretory phenotype

CoppĂŠ et al. Annu Rev Pathol 2010

The concept of « inflamm-aging »

Franceschi et al. Annals New York Academy of Science 2000

The concept of « inflamm-aging »

Franceschi et al. Annals New York Academy of Science 2000

•Claire Jacques (MCU) •Xavier Houard (MCU) •Jérémie Sellam (MCU-PH) •Audrey Pigenet (Tech) •Sabrina Priam (Thes.)

•Carole Bougaut (Post-doc) •Marjolaine Gosset