The role of microRNAs in monocyte and macrophage activation and effector function in rheumatoid arthritis

Mentor 1: Prof Iain B McInnes (University of Glasgow) Mentor 2: Prof Luke O’Neill (Trinity College Dublin) Mentor 3: Prof Foo Y Liew (University of Glasgow)

Mariola Stolarska

Division of Immunology, Infection and Inflammation, Glasgow Biomedical Research Centre, University of Glasgow

Support: Dr Andrew Hamilton (University of Glasgow) Dr Alirio Melendez (University of Glasgow)

Prof Constantino Pitzalis (Barts and The London School of Medicine)

Non-Clinical Career Development Fellowship 01/09/09-01/09/2014

Institute of Infection, Immunity & Inflammation,

MicroRNA in Rheumatology

Mariola Kurowska-Stolarska (msks1y@clinmed.gla.ac.uk)

50 years of the Centre for Rheumatic Diseases in Glasgow 22nd May 2015

Rheumatoid Arthritis Centre of Excellence

McInnes IB, 2012, NEJM

Epigenetics DNA methylation Histone modification MicroRNA network

Genetic susceptibility 101 loci (Nature 2013)

30-50% of RA do not respond to current treatment achieve long-lasting remission Mechanisms that need to be elucidated - development of autoimmunity treatment-refractory states

Priorities in RA

Strategy

Dendritic cells because they start everything

Macrophages because they drive chronic inflammation

One miR can regulate many mRNAs that often belong to one pathway

miR recognises complementary sequence in 3’UTR part of mRNA and induces its degradation

microRNA

post-transcriptional fine-tuners of biological pathways

microRNA

Degradation of mRNA

mRNA

Protein

microRNA inhibitor

Effector 1

Effector 2

TF

miR

The role of microRNAs in monocyte and macrophage activation and effector function in rheumatoid arthritis

Mentor 1: Prof Iain B McInnes (University of Glasgow) Mentor 2: Prof Luke O’Neill (Trinity College Dublin) Mentor 3: Prof Foo Y Liew (University of Glasgow)

Mariola Stolarska

Division of Immunology, Infection and Inflammation, Glasgow Biomedical Research Centre, University of Glasgow

Support: Dr Andrew Hamilton (University of Glasgow) Dr Alirio Melendez (University of Glasgow)

Prof Constantino Pitzalis (Barts and The London School of Medicine)

Non-Clinical Career Development Fellowship 01/09/09-01/09/2014

Are miRs deregulated in myeloid cells in RA?

What do they do?

Can we target them or pathways regulated by them?

TNF, IL-1, IL-6, chemokines

Visual art by Sebastian Kaulitzki, Germany

Proteases Immune

complexes

CD4+ T cells

PAR2

TLRs FcR

Integrins

lectins Oxysterols

PAMPs; DAMPs

LXR

Synovial fibroblasts

miR-155 miR-34a miR-22 miR-125a

miR-223

microRNA network of in RA synovial macrophage

miR155 miR34a miR22 miR125

miR223

TNF, IL-1, IL-6, chemokines

RA synovial macrophage

NEW PATHWAYS : Identifying miRs in RA cells

Visual art by Sebastian Kaulitzki, Germany

PB HC PB RA SF0

50000

100000

150000

200000

250000

Co

pie

s o

f 1

55

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6

co

pie

s o

f R

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5.25

5.00

4.75

4.50

4.25

4.00

3.75

3.50

Disease Activity Score 28-itemm

iR1

55

co

py

n

um

be

r lo

g

miR155 is up-regulated in blood and synovial fluid monocytes of RA patients and correlates with DAS28

Unpublished; A.Elemesmari, MKS ; Piccinini et al Cell Rep. 2012

miR-155 expression is induced by TLR ligands including endogenous ligand Tenascin- C and pro-inflammatory cytokines

RA synovium OA synovium

miR-155, 10x miR-155, 10x 40x 40x

Scramble, 10x

miR-155/CD68/merged

miR155 is overexpressed in RA synovial tissues macrophages compared to OA

M Kurowska-Stolarska PNAS 2011

miR155 drives RA macrophage activation by inhibiting SHIP-1

A.Elemesmari, MKS

EOA synovium RA synovium

SHIP-1/CD68/DAPI

SHIP1 mRNA in RA

SHIP-1 Effector 1

Effector 2

TF

miR TNF

TLR

+anti-miR155

Anti-miR155 inhibits TNFa

THERAPEUTIC TARGET: Anti-miR155 ameliorates arthritis

Unpublished, Brian Morton, Ashleigh Rainey, MKS

LPS (1ng/ml)

d41 d42 d1 d21

CFA+CII CII anti-miRs

d22 d36 d29

cull

Inflammation

Fatty acid synthesis

(Fasn)

LXRa

synovial mac; atherosclerotic plaque mac Kupffer cells

Cholesterol

uptake (CD36)

Triglyceride

metabolism (lpl)

Liver homeostasis

Socs1

Metabolic syndrome

miR-155

SHIP-1

TNF, IL-6, chemokines

bcl6

Chronic

inflammation

arthritis

NFkb

atherosclerosis

Kurowska-Stolarska & Alivernini PNAS 2011; S Bluml, A&R2011;

M Nazari-Jahantigh M, JCI 2012; AM. Piccinini, Cell Rep 2012;

A.Androulidaki, Immunity 2009; RM. O’Connell PNAS 2009; J.

Stanczyk, A&R 2008.

Down-regulation of miR-223 In RA macrophages facilitates production of IL-1b

Cel

ls a

lon

e

Co

ntr

l mim

ic

miR

-22

3 m

NLRP3

β-actin

Control mimic

miR-223 mimic

Control inhibitor

miR-223 inhibitor

Haneklaus et al JI 2012

miR-155 miR-34a miR-22 miR-125

miR-223

RA synovial macrophage

TNF, IL-1,IL-6, chemokines

miR155 miR223

SHIP-1/SOCS1

Inflammasone

IL-1,IL-18

SUMMARY: MicroRNAs integrating multiple activation signals

in RA macrophage

Acknowledgments

University of Glasgow, UK

Gianina Statache

Derek Gilchrist Aziza Elmesmari Lynn Stewart Donna McIntyre David Welsh

Clare Tange Brian Morton James Reilly Neal Millar Shauna Kerr Alasdair Fraser Ashley Miller Charles McSharry Iain McInnes

Gartnavel General Hospital and Royal Infirmary, Glasgow UK Duncan Porter John Hunter

Molecular Neurobiology Laboratory The Salk Institute Anna Zagorska Greg Lemke

Trinity College Dublin, IRL Luke O’Neill Seth Masters Institute of Rheumatology,

Catholic University of the Sacred Heart, Rome Stefano Alivernini

Carol Feghali-Bostwick

Masterswitch project

Medical University of South Carolina, Charleston