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Ulf Hedin Dept Vascular Surgery Karolinska University Hospital and Karolinska Institute Stockholm, Sweden Transcriptomic insights into carotid artery plaque vulnerability

Transcriptomic insights into carotid artery plaque vulnerabilitypast.mac-conference.com/xconfig/upload/files/$01-Fr_U. Hedin... · Ulf Hedin Dept Vascular Surgery Karolinska University

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Page 1: Transcriptomic insights into carotid artery plaque vulnerabilitypast.mac-conference.com/xconfig/upload/files/$01-Fr_U. Hedin... · Ulf Hedin Dept Vascular Surgery Karolinska University

Ulf HedinDept Vascular Surgery

Karolinska University Hospital and Karolinska Institute

Stockholm, Sweden

Transcriptomic insights into carotidartery plaque vulnerability

Page 2: Transcriptomic insights into carotid artery plaque vulnerabilitypast.mac-conference.com/xconfig/upload/files/$01-Fr_U. Hedin... · Ulf Hedin Dept Vascular Surgery Karolinska University

Ulf Hedin

FINANCIAL DISCLOSURE:

None

UNLABELED/UNAPPROVED USES DISCLOSURE:

None

Disclosures

Page 3: Transcriptomic insights into carotid artery plaque vulnerabilitypast.mac-conference.com/xconfig/upload/files/$01-Fr_U. Hedin... · Ulf Hedin Dept Vascular Surgery Karolinska University

Unstable or Stable?

CEA and Stroke prevention Unresolved issues

Stroke prevention today

Symptomatic patients

Presumably unstable carotid plaques

Asymptomatic patients

Presumably stable carotid plaques

Annual CEA at Karolinska

100 symptomatic patients = 17prevented strokes

10 asymptomatic patients = <0.5prevented stroke

We lack predictive tools…

Before MI After MI

Biomarkers

Page 4: Transcriptomic insights into carotid artery plaque vulnerabilitypast.mac-conference.com/xconfig/upload/files/$01-Fr_U. Hedin... · Ulf Hedin Dept Vascular Surgery Karolinska University

Aims

Understand plaque instabilityin humans

Identify

the vulnerable patient Circulating biomarkers

the unstable lesion Imaging

Improve prevention Plaque stabilizing drugs

Page 5: Transcriptomic insights into carotid artery plaque vulnerabilitypast.mac-conference.com/xconfig/upload/files/$01-Fr_U. Hedin... · Ulf Hedin Dept Vascular Surgery Karolinska University

Biobank of Karolinska Endarterectomies(BiKE)

Datasets/Resources

Clinical Platform

N=1000 patients• >100 clinical variables• SNP chip n=127• CTai=400• Duplex=600

Transcriptomics

Affymetrix arrays• n=127+10• n=50+5• n=40 high/low calcified

• n=96 PBMCs

Proteomics

Plaque: MSHIRiEF• n=40Plasma: Luminex• n=100 local• n=200 peripheral

Outcome• National registries• MI, Stroke, Intervention,

Death, Hospitalization

Analytical Platform

In situ analyses• HC/IHC/PLA

n=1000• LCM n=20

Exploratory Platform

In Vitro

In VivoAnimal Models

Transgenic/KO MiceRat Carotid Balloon Injury

Rat/Mouse Vein Graft ModelAneurysm Models

Zebra Fish

ValidationsPublic Databases

Collaborations

Page 6: Transcriptomic insights into carotid artery plaque vulnerabilitypast.mac-conference.com/xconfig/upload/files/$01-Fr_U. Hedin... · Ulf Hedin Dept Vascular Surgery Karolinska University

Analysing the molecular signatureSymptomatic vs. Asymptomatic patients

Transcript profile Affymetrix array; n>22 000 genes

Protein profile MSHiRIEF; n=9600 proteins GO function GO componentB C

T cell activation

innate immune response

adaptive immune response

B cell activation

macrophage activation

cytokine production

blood coagulation

interleukin-6 production

interferon-gamma signaling pathway

antigen processing

mast cell activation

apoptotic cell clearance

interleukin-1 secretion

response to interleukin-4

interleukin-10 production

A GO process

chemokine

activity

peptidase

activity

blood

coagulation

T-cells

activation

D

haptoglobin-hemoglobin complex

HDL p

articles

LDL par

ticlesECM

endocytic v

esicles

haptoglobin binding

MMP activit

yphosp

hatidylcholine

ster

ol ac

tivit

y

chemokine receptor

binding

cell proliferation

involved in wound

healingNO transport

cholester

ol ester

i cation

LDL par

ticle remodelling

lipopr

otein p

article

clearanc

e

ECM di

ssasse

mbly

Symptomatic Asymptomatic

Enrichment analysis Bioinformatic/computational processing

of massive data

Clustering of genes/proteins in enriched(up- or downregulated) categories/biological processes

Page 7: Transcriptomic insights into carotid artery plaque vulnerabilitypast.mac-conference.com/xconfig/upload/files/$01-Fr_U. Hedin... · Ulf Hedin Dept Vascular Surgery Karolinska University

Transcriptomic profilesSymptomatic vs. Asymptomatic patients

Enriched pathways/processes• Lipoprotein particle clearance

• Bone resorption

• Oxidative stress

• Immune system activation

• ECM disassembly

• Angiogenesis

• Foam-cell differentiation

Downregulatedpathways/processes

• Smooth muscle cell related pathways

• Proliferation, fibrosis

Ljubica MaticPerisic

Perisic L et al, J Intern Med. 2016;279:293-308.

Bone resorption=osteoclast activity=less calcification

• Less calcification in plaques from symptomatic patients???

Page 8: Transcriptomic insights into carotid artery plaque vulnerabilitypast.mac-conference.com/xconfig/upload/files/$01-Fr_U. Hedin... · Ulf Hedin Dept Vascular Surgery Karolinska University

Patient phenotypeStatins promote osteogenic processes/calcification in carotid plaques

CP vs Normal arteryn=127 n=10

Symptomatic (Unstable) vs. Asymptomatic (Stable)n=87 n=40

TIA/MS vs. AFn=61 n=26

>1 mo vs. <2 wkn=48 n=15

Statins Yes vs Non=73 n=14

Plaques from symptomatic patients without statins have enriched pathways

associated with less calcification

Plaques from symptomatic patients on statins have enriched pathways

associated with more calcification

Perisic L et al, J Intern Med. 2016;279:293-308.

Statins=reduce stroke risk

• Calcification part of the stabilizing effect of statins???

Diabetes Care 35:2390–2392, 2012

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Calcification vs. Plaque calcification

CVD Risk vs. Plaque instability

J Vasc Surg 2010;51:1015-25

Degree of plaque calcificat ion vs clinical ischemic symptoms

Sharma et al. Vascular Health and Risk Management 2010:6 603–611

Coronary calcification = Increased CVD risk

Carotid calcification =Decreased Stroke risk

Page 10: Transcriptomic insights into carotid artery plaque vulnerabilitypast.mac-conference.com/xconfig/upload/files/$01-Fr_U. Hedin... · Ulf Hedin Dept Vascular Surgery Karolinska University

BiKE

Patients n=40

CT scans

(TeraRecon)

High calcified >20% vs. low calcified <20%

Microarrays

Plaques

(Affymetrix HTA)

Bioinformatics

Gene Ontology

GSEA

Molecular signature ofcarotid calcification? Eva Karlöf Lars

MaegdefesselLjubica Matic

Perisic

Nuno Dias

Page 11: Transcriptomic insights into carotid artery plaque vulnerabilitypast.mac-conference.com/xconfig/upload/files/$01-Fr_U. Hedin... · Ulf Hedin Dept Vascular Surgery Karolinska University

P<0.05

Karlof E et al, unpublished data

Up/Down regulated processes

developmental process

muscle system process

anatomical structure

morphogenesis

regulation of muscle

contraction

cell adhesion

Upregulated processes

high vs. low calcified plaques

(GSEA)

Enriched pathways/processes

• Smooth muscle related pathways

• Remodelling

• Proliferation

• Fibrosis

Downregulatedpathways/processes

• Immune response

• Cholesterol/lipid transport

• Leukocyte migration

• Protease activity/matrix degradation

Carotid calcification =Stable molecular signature

Page 12: Transcriptomic insights into carotid artery plaque vulnerabilitypast.mac-conference.com/xconfig/upload/files/$01-Fr_U. Hedin... · Ulf Hedin Dept Vascular Surgery Karolinska University

Karlof E et al, unpublished data

Dysregulated genes in high vs lowcalcified plaques

Page 13: Transcriptomic insights into carotid artery plaque vulnerabilitypast.mac-conference.com/xconfig/upload/files/$01-Fr_U. Hedin... · Ulf Hedin Dept Vascular Surgery Karolinska University

Summary The molecular signature of the

unstable carotid atheroma

Provides…

Target validation in human atherosclerosis

New hypotheses and noveltarget processes/pathways

Suggests…

Plaque calcification as a stabilizing feature

Statins promote calcification

Smooth muscle cell differentiation in association with calcification

Carotid calcification (by Ctai) predict low stroke risk

SMCs

Calcification Lipids

Inflammation

Stable Unstable

Page 14: Transcriptomic insights into carotid artery plaque vulnerabilitypast.mac-conference.com/xconfig/upload/files/$01-Fr_U. Hedin... · Ulf Hedin Dept Vascular Surgery Karolinska University

Ljubica Matic PerisicEva KarlöfUrsula RykazchewskaAnton RazuvaevJoy RoySilvia AldiMariette LengquistMalin KronqvistSiw FrebeliusErika HedinOlga NilssonPeter Gillgren

CMMGöran HanssonGabrielle BerneLars MaegdefesselAnders HamstenPer ErikssonMaria Sabater LlealSci LifeJacob OdebergMaria Jesus IglesiasFredrik PonténJanne LehtiöMatthias Vesterlund

Stanford Tom Querthermos LabNick Leeper LabOttawa Heart InstituteRuth Mc Perason LabKatey Rainer LabKing’s College Manuel Mayer LabFrankfurtStephanie Dimmeler LabKostas Stellos LabLund/MalmöNuno Dias

Göran Gabrielle

Ljubica

AntonSiw

MalinMette

Joy

Olga