Download Immune Cell Ac va on Among Lung Adenocarcinoma and

ImmuneCellAc@va@onAmongLungAdenocarcinomaandSquamousCell
CarcinomaSubtypesandCD274(PD-L1)Expression
FarukiH1,MayhewG1,HayesN2,SerodyJ2,PerouC2,3,Lai-Goldman,M1,(1)GeneCentric,Durham,NC(2)LinebergerCancerCenter,(3)DepartmentofGeneAcs,UniversityofNorthCarolina,ChapelHill,NC
Th1 cells(AI)
Th2 cells(AI)
TFH(AI)
Th17 cells(AI)
Tgd(AI)
Cytotoxic cells(AI)
NK cells(II)
NK CD56dim cells(II)
NK CD56bright cells(II)
DC(II)
iDC(II)
aDC(II)
pDC(II)
Eosinophils(II)
(NE)
Mast cells(II)
Neutrophils(II)
Adenocarcinoma
(AC)
IFN
PDL1
PDCD1
CTLA4
Squamoid
BiologicSubtypes
PROXIMAL
INFLAMMATORY
[ 5 ] CONFIDENTIAL AND PROPRIETARY
Tgd(AI)
Cytotoxic cells(AI)
NK CD56dim cells(II)
NK CD56bright cells(II)
DC(II)
Eosinophils(II)
Mast cells(II)
Neutrophils(II)
IFN
PDL2
PDCD1
CTLA4
T helper cells
0.5
Tem
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(magnoid)
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PP
(magnoid)
PI
(squamoid)
●
0.30
0.35
0.30
0.25
0.20
0.00
0.35
0.05
0.10
0.15
0.20
0.25
0.30
0.35
Subtype Association Strength
SquamousCellCarcinoma(TCGA,UNC,Raponi)
DC
secretory
primitive
classical
basal
overall
Th2 cells
DC
TFH
iDC
Th17 cells
Eosinophils
PDL1
CD8 T cells
Macrophages
CTLA4
TFH
iDC
TReg
Neutrophils
TRU
(bronchioid)
PP
●
(magnoid)
PI
(squamoid)
TRU
(bronchioid)
1.0
1.5
2.0
0.5
1.0
1.5
2.0
HR, 95% CI
IICHR,95%CI
0.5
1.0
1.5
2.0
HR, 95% CI
SinglegeneHR,95%CI
0.5
1.0
1.5
2.0
HR, 95% CI
AICHR,95%CI
0.5
1.0
1.5
2.0
HR, 95% CI
IICHR,95%CI
CONCLUSIONS
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0.25
PI (squamoid)
PP (magnoid)
TRU (bronchioid)
overall
HR, 95% CI
AICHR,95%CI
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NK CD56bright cells(IIC)
Th17 cells(AIC)
PDL1
PDL2
Tgd(AIC)
Th2 cells(AIC)
IFN
aDC(IIC)
T helper cells(AIC)
B cells(AIC)
TFH(AIC)
Th1 cells(AIC)
CTLA4
TReg(AIC)
NK CD56dim cells(IIC)
CD8 T cells(AIC)
PDCD1
Cytotoxic cells(AIC)
T cells(AIC)
Neutrophils(IIC)
pDC(IIC)
iDC(IIC)
DC(IIC)
Macrophages(IIC)
Eosinophils(IIC)
Mast cells(IIC)
NK cells(IIC)
Tcm(AIC)
Tem(AIC)
Th17 cells(AIC)
Tgd(AIC)
Th2 cells(AIC)
Tcm(AIC)
Tem(AIC)
pDC(IIC)
Eosinophils(IIC)
Mast cells(IIC)
Neutrophils(IIC)
Macrophages(IIC)
DC(IIC)
iDC(IIC)
T helper cells(AIC)
NK CD56dim cells(IIC)
TReg(AIC)
CTLA4
Th1 cells(AIC)
PDCD1
CD8 T cells(AIC)
Cytotoxic cells(AIC)
T cells(AIC)
PDL1
PDL2
IFN
aDC(IIC)
NK CD56bright cells(IIC)
NK cells(IIC)
B cells(AIC)
TFH(AIC)
2
1
0
T.cells
4A.
PP
●
(magnoid)
PI
(squamoid)
•  LungADandSQgeneexpressionsubtypesvaryintheirimmunelandscape.
•  AdenocarcinomaPP(magnoid)subtypeandSQclassicalsubtypeshow
minimalimmuneinfiltraAonsuggesAngreducedresponsetoimmunoRx.
•  InSQ,subtypeappearstobeabederpredictorofimmuneinfiltraAonthan
CD274(PD-L1).CD274expressionwasnotassociatedwithAICexpression
norwithimprovedsurvivalinSQ.
•  AdenocarcinomaPIsubtype(squamoid)andtheSQprimiAvesubtype
showedimmunefeatureexpressionassociatedwithimprovedsurvival.
•  LungAD&SQsubtypesmaybehelpfulaspotenAalimmunoRxbiomarkers.
●
Affymetrixn=130UNCAgilentn=56TCGASQRNAseq
4n=501
T.cells
T.cells
T.cells
T.cells
LeeAffymetrixn=75Raponi
LEE SQ n=75 p=3.1e−06
RAP SQ n=130 p=2.4e−06
UNC SQ n=56 p=0.00022
TCGA SQ n=501 p=1.1e−27
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basal
secretory
primitive
●
classical
basal
secretory
primitive
−2
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basal
−2
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classical
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basal
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secretory
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primitive
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4B.
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T.cells
Characteris@c
TCGA4
Lee8
Raponi9
UNC3
Total#ofsamples
501
75
130
56
TissuepreservaAon FreshFrozen FreshFrozen FreshFrozen FreshFrozen
Subtype
Basal
137
13
35
12
Classical
170
31
42
21
PrimiAve
73
12
23
9
Secretory
121
19
30
14
Stage
I
241
0
73
34
II
152
0
33
19
III
85
0
23
3
IV
7
0
0
0
StageNA
16
75
0
0
1.0
0.0
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0.10
B cells
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Table2.LungSquamousCellCarcinomaDatasets
TCGA SQ n=501
●
● ●
0.05
Adenocarcinoma(TCGA,Shedden,&Tomida)
●
●
●
(HR)andconfidenceintervalscalculatedfromstraAfiedcoxmodels.Subtype
specificHR’sareadjustedforstage(overalladjustedbystageandsubtype)and
onlyimmunefeatureswithsignificantassociaAons(p<0.05)areshown.
PDL1
T.cells
T.cells
T.cells
T.cells
T.cells
TCGARNAseqn=515SheddenAffyn=442TomidaAgilentn=117UNCAgilentn=116GeneCentricFFPEn=88
TCGA AD n=515 p=2.7e−16
SHEDDEN AD n=442 p=2.1e−16
TOMIDA AD n=117 p=1.4e−06
UNC AD n=116 p=6.7e−08
SEATTLE AD n=88 p=0.00026
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Figure6.Signature-survivalassociaAonsoverallandbysubtype.HazardRaAos
Macrophages(II)
mulApleADdatasets(A)andSQdatasets(B).
●
●
Subtype Association Strength
Th17 cells(AIC)
NK CD56bright cells(IIC)
●
●
●
●
pDC(II)
Figure4.ReproducibilityofTcellsignaturegeneexpressionsubtypepadernsacross
−1
Characteris@c
TCGA2
Shedden6
Tomida7
UNC1
GeneCentric
Total#ofsamples
515
442
117
116
88
TissuepreservaAon FreshFrozen FreshFrozen FreshFrozen FreshFrozen
FFPE
Subtype
TRU(Bronchioid)
188
159
45
47
26
PP(Magnoid)
154
161
40
40
29
PI(Squamoid)
173
122
32
29
33
Stage
StageI
276
276
79
73
42
StageII
123
95
13
21
7
StageIII
84
68
25
19
15
StageIV
27
0
0
2
24
StageNA
5
3
0
1
0
●
aDC(II)
wheresignaturesarearrangedbyhierarchicalclustering.
UsingpreviouslypublishedBindeaetal.(5)immunecellgenesignatures(24intotal)and
TCGA AD n=515
ADandSQsubtypinggeneexpressionsignatures(1-4),severalpublicallyavailablelung
Tem(AIC)
TFH(AIC)
Tcm(AIC)
ADandSQdatasets(2,4-9),wereexaminedforimmunecellfeaturesinrelaAontoAD
B cells(AIC)
NK cells(IIC)
NK cells(IIC)
Mast cells(IIC)
NK CD56bright cells(IIC)
andSQsubtypes.GeneexpressionsignaturesofbothInnateImmuneCells(IIC)and
Eosinophils(IIC)
aDC(IIC)
Macrophages(IIC)
IFN
DC(IIC)
AdapAveImmuneCells(AIC),a13geneIFNsignature(IFN),aswellassinglegene
PDL2
iDC(IIC)
PDL1
pDC(IIC)
T cells(AIC)
Neutrophils(IIC)
immune-biomarkers(CTLA4,PDCD1,andCD274(PD-L1),PDCDLG2(PD-L2))were
Cytotoxic cells(AIC)
T cells(AIC)
CD8 T cells(AIC)
Cytotoxic cells(AIC)
PDCD1
examinedinthe3ADsubtypes(TRU,PP,andPI)andthe4SQsubtypes(PrimiAve,
PDCD1
Th1 cells(AIC)
CD8 T cells(AIC)
CTLA4
NK CD56dim cells(IIC)
TReg(AIC)
Classical,Secretory,Basal).ImmunecellsignatureassociaAonswithtumorsubtypeand
TReg(AIC)
NK CD56dim cells(IIC)
CTLA4
T helper cells(AIC)
Th1 cells(AIC)
withCD274expressionwereevaluatedusinglinearregression.Hierarchicalclusteringof
iDC(IIC)
TFH(AIC)
DC(IIC)
B cells(AIC)
Macrophages(IIC)
T helper cells(AIC)
immunesignaturesandpairwisesignaturecorrelaAonswerealsoanalyzed.SurvivalNeutrophils(IIC)
aDC(IIC)
Mast cells(IIC)
IFN
Eosinophils(IIC)
signatureassociaAonsofStagesI-IIIsampleswereevaluatedwithstraAfiedcox
Th2 cells(AIC)
pDC(IIC)
Tgd(AIC)
Tem(AIC)
PDL2
proporAonalhazardmodelsallowingfordifferentbaselinehazardsineachdataset.
Tcm(AIC)
PDL1
Th2 cells(AIC)
RESULTS
Table1.LungAdenocarcinomaDatasets
●
0.00
iDC(II)
Figure3.CorrelaAonmatricesofimmunecellsignaturesintheTCGAADandSQdataset
Tgd(AIC)
Th17 cells(AIC)
●
NK cells(II)
2
PROXIMAL
PROLIFERATIVE
●
●
CD8 T cells(AI)
1
Magnoid
TERMINAL
RESPIRATORY
TReg(AI)
0
Bronchioid
Th17 cells(AI)
T.cells
METHODS
Basal
Secretory
TFH(AI)
−1
Classical
Th2 cells(AI)
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secretory
Primitive
Th1 cells(AI)
primitive
(SCLC)
Tem(AI)
Individualgenes
PDL2
Carcinoid
Tcm(AI)
classical
Small Cell
Lung Cancer
T helper cells(AI)
OtherInnatecells
Interferonsignature
Macrophages(II)
Squamous Cell
Carcinoma (SCC)
T cells(AI)
B cells
T cells
T helper cells
Tcm
Tem
Th1 cells
Th2 cells
TFH
Th17 cells
TReg
CD8 T cells
Tgd
Cytotoxic cells
0.15
Tem(AI)
Neuroendocrine
Subtype
B cells(AI)
●
0.10
Tcm(AI)
CD8 T cells(AI)
(NSCLC)
1.5
0.05
T helper cells(AI)
TReg(AI)
Non-Small Cell
Lung Cancer
1.0
Bcells
Tcells
NKcells
DendriAccells
T cells(AI)
Lung Cancer
0.5
0.00
Subtype
B cells(AI)
Lung Cancer Subtyping
−0.5
PDL1 Association Strength
−1.5
0.35
0.5 1.0 1.5
0.30
−0.5
0.25
−1.5
i d )
o
oi d )
n
g
m
a
a
(m
sq u
ŸPP
ŸPI(
TCGA SQ n=501
TCGA AD n=515
0.20
TCGASQn=501
0.15
●
0.05
TCGAADn=515 ●
TCGA SQ n=501
bronchioid
magnoid
squamoid
0.00
●
PDL1 Association Strength
TCGA AD n=515
1
GeneexpressionbasedsubtypinghasconsistentlyidenAfied3disAnct
biologicsubtypesinLungAdenocarcinoma(AD),TerminalRespiratoryUnit
(TRU)formerlyBronchioid,ProximalProliferaAve(PP)formerlyMagnoid,
andProximalInflammatory(PI)formerlySquamoid(1,2)and4subtypes
withinSQ,PrimiAve,Classical,BasalandSecretory(3,4)(SeeFigure1).AD
andSQsubtypesdemonstratekeydifferencesingenomicalteraAons,
tumordrivers,prognosis,andlikelyresponsetovarioustherapies(1-4).
andAICsignaturesversussubtypeandAICsignatures.InSQ,associaAonis
consistentlygreaterforsubtypesthanforPD-L1.
markersintheTCGALungADandSQdatasets.
1
BACKGROUND
Figure1.
Figure5.AssociaAon(adjustedR-squared)betweenCD274(PD-L1)expression
Figure2.HeatmapsofBindeaetal.(5)immunecellsignaturesandotherimmune
0.10
●
REFERENCES
1.  WilkersonMD,etal.PLoSOne2012;7(5):e36530.PMID22590557
2.  TCGALungAdenoC.Nature2014;511(7511):543-550.PMID25079552
3.  WilkersonMD,etal.ClinCancerRes2010;16(19):4864-75.PMID20643781
4.  TCGALungSQCC.Nature2012;489(7417):519-525.PMID22960745
5.  Bindeaetal.Immunity2013;39(4):782-95.PMID24138885
6.  SheddenK.etal.NatMed2008;14(8):822-827.PMID18641660
7.  TomidaS.etal.JClinOncol2009;27(17):2793-99.PMID19414676
8.  LeeES,etal.CancerRes2008;14(22):7397-7404.PMID19010856
9.  RaponiM,etal.CancerRes2006;66(7):466-72.PMID16885343
ABBREVIATIONS
AD=Adenocarcinoma
SQ=Squamouscellcarcinoma
TRU=TerminalRespiratoryUnit
PP=ProximalProliferaAve
PI=ProximalInflammatory
AIC=AdapAveImmuneCells
IIC=InnateImmuneCells
HR=HazardRaAo
Abstract#3077.PresentedattheAmericanSocietyforClinicalOncology(ASCO)AnnualMeeAng.Chicago,IL.June3-7,2016.Emailcontact:[email protected]