Marginal Zone Lymphoma Soma2c muta2ons and mechanisms. DU.pdfBIR 3 BIR 1 BIR 2 Ig BIR 3 BIR 1 BIR 2...

MarginalZoneLymphomaSoma2cmuta2onsandmechanisms

Ming-Qing Du Department of Pathology, University of Cambridge

� Immunologicals2mula2ons

� gene2cchanges� oncogeniccoopera2on

SignallinginMarginalZoneB-cells

NOTCH

BCR

TLR

BAFFR

CD40

Geneticchanges

Geneticchanges

Geneticchanges

Geneticchanges

MALTlymphomaconceptChronicinflammatorydisorders

Mucosalsiteswithoutna2veMALT

LowgradeMALTlymphoma

MALTlymphoma Ae2ology

Stomach Hpylori ~95%

Skin Borreliaburgdorferi some

Ocularadnexa ChlamydiapsiQaci variable

Lung AXyloxidans variable

IPSID Campylobacterjejuni variable

Salivaryglands Sjogren’ssyndrome/HCV everycase

Thyroid Hashimotothyroidi2s everycase

Responsetoan2-microbialtherapy

CRin~70%

CRincasereport

CRin~20%;PRin~55%

CRincasereport

SMZL HCV variable

NMZL HCV variable

ORRin~50%

CRin~70%

o SurfaceIgM+

o Proliferateuponcross-linkingsIg.

o Hyper-soma2candongoingmuta2onsinIGH/IGL.

o TumourIgsfromgastricMALTlymphomadonotrecognisingHpylorian2gen,butpolyreac2vetoauto-an2gens.

An2genics2mula2on:BCRsignalling

Husselletal,JPathol1993DuetalLeukaemia1996&Blood1996

HusselletalAmJPathol1993CraigetalBlood2010)

target genes Nucleus

BCR

p50 p65IKBPK48Ub

p50 p65

CARD11

BCL10

MALT1

autoan;gen

BiasedIGgeneusageinMALTlymphomaMALTlymphoma BiasedIGgeneusage

StomachIGHV3-7IGHV1-69

Salivaryglands IGHV1-69/IGKV3-20(~50%)

Moodyetalunpublished;DuMQ,SeminsCancerBiol2016

KnownIGproper2es

� rheumatoidfactors� self-polyreac2ve

Sample VGene JGeneCDR3Length CDR3sequence

RF-112/113 IGHV1-69 JH4 13 E G R S S D Y S N P F D YRF-BOR IGHV1-69 JH4 12 E G R RM A I - N P F D YSA_002 IGHV1-69 JH4 12 E G QQM S T - N P F D FSA_006 IGHV1-69 JH4 13 E G K A T V T T N P F D YSA_010 IGHV1-69 JH4 12 E G R QM P T - N P F D YSA_6T IGHV1-69 JH4 13 E G K S S D Y S N P F D YD26010 IGHV1-69 JH4 12 E G R Q T V T - N P F D Y

SalivaryglandMALT-L

RF

Ocularadnexa IGHV4-34/IGK3-20(~30%)

Thyroid IGHV3-30?

� bindstoN-acetyl-lactosamine� polyreac2ve� self-polyreac2ve

An2genics2mula2on:Tcellhelp

HusselletalLancet1993


BCR

p100

p52RelB

RelBp50 p65IKBPK48Ub

p50 p65

CARD11

BCL10

MALT1

autoan;gen

Reac;vefollicle

Tumourcellarea

HpylorispecificT–cells� polyclonal� bystanderhelps� cytokines/solubleligands

?OtherB-cellhelpsignals


BCR

p100

p52RelB

RelBp50 p65IKBPK48Ub

p50 p65

CARD11

BCL10

MALT1

autoan;gen

Hpylorieradica2on:assumedeffect

70%gastricMALTlymphomacanbecuredbyHpylorieradica2onalone

Gene2cchangesinMALTlymphoma

Liuetal,Gastroenterology2002,Yeatal,Blood2001,2003,Yeatal,JPathol2005,GoatlyetalModernPathol2008Chanudetetal,Leukaemia2010Bietal,Haematologica2012

0%

20%

40%

60%

t(14;18)/IGH-MALT1

t(1;14)/IGH-BCL10

t(11;18)/API2-MALT1

A20inactivation

MYD88mutation

A20(TNFAIP3)inactivation


p50 p65IKBPK48Ub

p50 p65

NEMO IKKβ IKKα P

K63Ub

BCL10

MALT1

DD

Ig

Ig

CARD

BCL10

MALT1

DD

Ig

Ig

Casp-L

CARD

Casp-L

t(1;14)IGH-BCL10

t(14;18)IGH-MALT1

t(11;18)API2-MALT1

Ig

BIR3

BIR1

BIR2 Ig

BIR3

BIR1

BIR2

p100

p52RelB

RelB

IKKα IKKα

NIK

K63Ub

TAB2 TAK1 P

TRAF6

A20

API2

93%2% 1%E6 E7 E8 E9

BIR BIR CARD RINGBIR

32% 17%42%9%E3 E9E8E5

4%

MALT1 DD Ig-L Caspase-likeIg-L

Binds to BCL10

Ig-L

t(11;18)

E3 ligaseHeterotypical oligomerisation

Proteolytic activity

UBA

LIMA1

LMO

Oncogenicproper2es

NEMO

IKKβ IKKα

IKKα IKKα

NIK

p100

p52RelB

Target genesNucleus

Proteasome

RelB

P

K63Ub

p65p50

TAB2

TAK1 P

TRAF6

K63Ub

A20p50 p65

IKBPK48Ub

MALT1API2-MALT1

NEMO

IKKβ IKKα

IKKα IKKα

NIK

p100

p52RelB

Target genesNucleus

Proteasome

RelB

P

K63Ub

p65p50

TAB2

TAK1 P

TRAF6

K63Ub

A20p50 p65

IKBPK48Ub

MALT1API2-MALT1

NEMO

IKKβ IKKα

IKKα IKKα

NIK

p100

p52RelB

Target genesNucleus

Proteasome

RelB

P

K63Ub

p65p50

TAB2

TAK1 P

TRAF6

K63Ub

A20p50 p65

IKBPK48Ub

MALT1API2-MALT1

DuMQ,SeminsCancerBiol2016RosebecketalScience2011NieetalNatCommun2015

DuMQ,SeminsCancerBiol2016HamoudietlLeukemia2010

BCR

autoan;gen

BCL2 Apoptosis

CCR2

MAPK

BAFFRCD40TLR6

TLR6

target genesNucleus

p50 p65IKBPK48Ub

p50 p65

NEMOIKKβ IKKαP

K63Ub

p100

p52RelB

RelB

IKKα IKKα

NIK

K63Ub

TAB2 TAK1 P

TRAF6

A20

LIMA1

LIM

Oncogenicproperties

NEMO

IKKβ IKKα

IKKα IKKα

NIK

p100

p52RelB

Target genesNucleus

Proteasome

RelB

P

K63Ub

p65p50

TAB2

TAK1 P

TRAF6

K63Ub

A20p50 p65

IKBPK48Ub

MALT1API2-MALT1

NEMO

IKKβ IKKα

IKKα IKKα

NIK

p100

p52RelB

Target genesNucleus

Proteasome

RelB

P

K63Ub

p65p50

TAB2

TAK1 P

TRAF6

K63Ub

A20p50 p65

IKBPK48Ub

MALT1API2-MALT1

NEMO

IKKβ IKKα

IKKα IKKα

NIK

p100

p52RelB

Target genesNucleus

Proteasome

RelB

P

K63Ub

p65p50

TAB2

TAK1 P

TRAF6

K63Ub

A20p50 p65

IKBPK48Ub

MALT1API2-MALT1

BCL10

MALT1 API2-MALT1

LMO

Complete remission No or partial remission Europe No evidence of hRD Presence of hRD stage I ≥ stage II

Apen et al Blood 2000 0/18 0% Liu et al Gastroenterology 2002 0/46 0% 2/2 26/43 60% 16/20 80%

Wundisch et al JCO 2005 3/46 7% 4/7 57% 3/13 23%

Montalban et al Ann Oncology 2005 0/11 0% 0/2 0% Levy, et al JCO 2005 0/21 0% 1/7 14% 2/3 67% Ruskone-Fourmestraux, Du et al unpublished 2/24 8% 1/2 50% 14/26 54% 8/9 89%

Subtotal 5/145 3% 7/11 64% 44/91 48%% 26/32 81%

Far East Nakamura et al JCP 2003 0/11 0% 0/4 0% 0/4 0% Iwano et al J Gastroenterology 2004 0/5 0% 1/4 25% 2/2 100% Yeh et al Blood 2005 1/36 2.7 12/23 52% Nakamura et al Gut 2007 1/20 5% 5/14 36% Nakamura et al AM J Gastr 2007 1/57 2% 10/21 47% 4/9 44%

Subtotal 3/129 2% 28/66 42% 6/15 40% Total 8/274 3% 71/150 47% 30/44 68%

T(11;18): a marker for gastric MALT lymphoma not responding to H. pylori eradication

MALTlymphomalacksothermuta2onsfrequentlyseeninABC-DLBCL

30%

2% 1%5%

3% 2% 2% 1% 2% 1% 1%0

5

10

15

20

25

30

35

% c

ases

mut

ated

K63 deubiquitinase E3 ubiquitin ligase (K48)

OTU: Ovarian tumour domain that belong to family of deubiquitinating cysteine proteases; ZF: zinc finger

deletion (39.7%) insertion (18%)nonsense mutation (25.9%) missense mutation (13.8% ) splicing site mutation (2.6% )

Previously reported

Identified in this study

OTU ZF ZF ZF ZF ZF ZF ZF

Identified in previous studies

Identified in OA-MALT-L

A20

Bietal,Haematologica2012,Escudero-IbarzetalHaematologica2016,JohanssonetalOncotarget20017;JungetalOncotarget2017,Moodyetal,Unpublisheddata

BiasedusageofIGgenesinOA-MALTlymphoma

0

5

10

15

20 IGHV4-34P=<0.001OA-MALT-L

IgMmemoryB-cells

FR1 CDR1 FR2 CDR2 FR3 . V4-34*02 QVQLQQWGAGLLKPSETLSLTCAVY GGSFSGYY WSWIRQPPGKGLEWIGE INHSGST NYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYC V4-34*01 QVQLQQWGAGLLKPSETLSLTCAVY GGSFSGYY WSWIRQPPGKGLEWIGE INHSGST NYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYC OA_019 ----R-----------------E-- -----D-- -----K----------- -----G- ------Q---V--I----K--Y----A----------- OA_029 --K-----PR--EA----------- -----DFS ------T---------D V--D-R- HH----TG-L---A--------IE-N------------ OA_047 ----H-------------C------ -----D-- ---V--S----P----- --DG-N- -------------I-A-------N-E------------ OA_086 --------TR--------------- ----TS-- ----------------- ------- -----------------------N-T--------M--- GA_010 ------------------------- ---L---- ----------------- ------- ---R-------------------N-N-M------I--- GA_011 --L-----S-VV---QS-------- -E----V- -G-V--TSR-R--C--- AS-T-II ------RG-AS---------V--S-T-L----SG--F- OA_017 ------------------------- -------- ----------------- ------- -------------------------------------- OA_044 -------------A----------- -----D-- -T--------------- --DA--- -----------------NT----R----------F--- OA_052 ------------------------- -D------ --------E-------- ---A-T- -------------S---------N-T--------M--- OA_088 ----H---------------M---- -------- -G--------------- --Y---- --------------------------------------




MaintaintheQW/AVY“hydrophobicpatch”requiredforbindingtoN-acetyl-lactosamine

Muta2onhotspots:N-glycosyla2onsite(60%)KLSmo2f(70%)

ZhuetalPlosOne2011DagklisetalLeukaemia2012

ZhuetalLeukaemia2015vanMaldegenmetalLeukaemia2012

Moodyetalunpublished

Significantassocia2onbetweenA20muta2onandIGHV4-34usageinOA-MALTlymphoma

Moodyetal,unpublisheddata

IGHV1-69 IGHV3-7 IGHV4-34 IGHV7-4-1 OtherIGHV

0%

25%

54%

0%

22%

0

10

20

30

40

50

60

%caseswith

A20

inac2va2

on

P=0.01**

P=0.19ns

OcularadnexalMALTlymphoma

SalivaryglandMALTlymphoma

Nucleus

p65 p50 p52

RelB

NOTCH

RBPJ

NICD

γ-secretase

BCR (IGHV4-34)

TLR

CARD11

BCL10

MALT1

p50 p65 IKB P

NEMO IKKβ IKKα

K48Ub

P

MYD88

IRAK4

IRAK1

K63Ub

p65 p50

TAB2 TAK1 P

TRAF6

K63Ub

A20

XA20

XA20

CD40, BAFFR

IKKα IKKα

NIK

p100

p52 RelB

RelB

autoan;gen

Moodyetal,unpublisheddataJohanssonetalOncotarget2016

8%

0

5

10

15

20

25

30

35

40

45

IGHV1-2 7qdeletion KLF2mutation

NOTCH2mutation

TRAF3mutation

TNFAIP3mutation

BCRpathwaymutation

MYD88mutation

TP53mutation

%ca

sesw

ithgen

eticch

ange

CaseswithKLF2mutation Caseswithout KLF2mutation CaseswithunknownKLF2status

KLF2muta2onandIGHV1-2usage

P=7.33x10-7P=1.92x10-7

0

5

10

15

20

25

30

35

40

45

IGHV1-2 7qdeletion KLF2mutation

NOTCH2mutation

TRAF3mutation

TNFAIP3mutation

BCRpathwaymutation

MYD88mutation

TP53mutation

%ca

sesw

ithgen

eticch

ange

CaseswithKLF2mutation Caseswithout KLF2mutation CaseswithunknownKLF2statusClipsonetalLeukaemia2015

BikosetalLeukemia2012WarsameetalBlood2011

IGHV1-2characteris2cs� over-represented(~30%)inSMZL

� mostbeingallele*04

� minimalsoma2cmuta2on

� longCDR3withcommonmo2f

� oyenassociatedwithbiaseduseoflightchaingenes

� polyreac2vetoautoan2genbyrecombinantan2bodystudies

XNucleus

p65 p50 p52

RelB

NOTCH2

RBPJ

NICD

γ-secretase

KLF2

TLR

CARD11

BCL10

MALT1

p50 p65 IKB P

NEMO IKKβ IKKα

K48Ub

P

MyD88

IRAK4

IRAK1

K63Ub

p65 p50

TAB2 TAK1 P

TRAF6

K63Ub

A20

CD40, BAFFR

IKKα IKKα

NIK

p100

p52 RelB

RelB

autoan;gen

BCR (IGHV1-2)

XTRAF2 IAP1/2

TRAF3

MarginalZonelymphoma

NOTCH

BCR

TLR

BAFFR

CD40

A20

BCL10

MALT1

API2-MALT1

MYD88

KLF2

NOTCH2

IGHV4-34IGHV1-2……

DepartmentofPathologyUniversityofCambridgeAlexandraClipsonMingWangSarahMoodyLeireEscuderoIbarzEguzkineOchoaWenqingYaoJoeThompson

Acknowledgements

Ins2tutUniversitaireDePathologie,LausanneLaurencedeLeval

WellcomeTrustSangerIns2tuteGeorgeVassiliou

Ex-labmemberRifatHamoudiAJamesWatkinsYingwenBiQingguoYanNaiyanZengEstelleChanudetXueminXue

UCLMedicalSchoolPeterIsaacson

MolecularMalignancyLaboratoryAddenbrokesHospitalHongxiangLiu

RoyalMarsdenHospital,LondonAndrewWotherspoon

SouthamptonUniversityHospitalsNHSTrustMargaretAshton-Key

MedicalUniversityofViennaMarkusRaderer

RoyalBournemouthHospital,BournemouthDavidOscier

King'sCollegeLondonDeborahDunn-Walters

DepartmentofHaematology,Addenbrooke’sHospitalGeorgeFollowsMikeScoQ

Chi-MeiMedicalCentre,TaiwanShih-SungChuang

StateHospitalSt.Gallen,SwitzerlandSergioCoglia{

WesternGeneralHospital,EdinburghJohnGoodlad

Marginal Zone Lymphoma Soma2c muta2ons and mechanisms. DU.pdfBIR 3 BIR 1 BIR 2 Ig BIR 3 BIR 1 BIR 2...

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