Transcriptomic insights into carotid artery plaque...
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Transcript of Transcriptomic insights into carotid artery plaque...
Ulf HedinDept Vascular Surgery
Karolinska University Hospital and Karolinska Institute
Stockholm, Sweden
Transcriptomic insights into carotidartery plaque vulnerability
Ulf Hedin
FINANCIAL DISCLOSURE:
None
UNLABELED/UNAPPROVED USES DISCLOSURE:
None
Disclosures
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
Aims
Understand plaque instabilityin humans
Identify
the vulnerable patient Circulating biomarkers
the unstable lesion Imaging
Improve prevention Plaque stabilizing drugs
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
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
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???
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
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
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
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
Karlof E et al, unpublished data
Dysregulated genes in high vs lowcalcified plaques
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
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