Linfoma di BurkittLinfoma di Burkitt

((BurkittBurkitt))

AimsAims

…….Attitudes are more .Attitudes are more important than abilitiesimportant than abilities

Motives are more Motives are more important than methodsimportant than methods

Character is moreCharacter is moreimportant than clevernessimportant than cleverness

And heart takes precedence And heart takes precedence over the headover the head……....

Denis P. Burkitt, 1991Denis P. Burkitt, 1991

LINFOMA DI BURKITT (LBT)LINFOMA DI BURKITT (LBT)

endemico: endemico: zone centrali dellzone centrali dell’’Africa (malaria)Africa (malaria)etetàà pediatrica; sedi: mandibola, pediatrica; sedi: mandibola, ovaioovaio

sporadico:sporadico: paesi occidentalipaesi occidentalietetàà variabile; sedi: intestino, variabile; sedi: intestino, gonadi, SNCgonadi, SNC

HIVHIV--correlatocorrelato:: SNC, etc.SNC, etc.

Buona risposta in etBuona risposta in etàà pediatrica con terapie aggressivepediatrica con terapie aggressive

FORMA ENDEMICAFORMA ENDEMICA(Lancet Oncology 2004; 5:738(Lancet Oncology 2004; 5:738--46)46)

Patogenesi del linfoma di Burkitt endemico Patogenesi del linfoma di Burkitt endemico legata alla cooperazione di:legata alla cooperazione di:-- EBVEBV (immortalizzazione),(immortalizzazione),-- malariamalaria (spinta proliferativa),(spinta proliferativa),-- ArbovirusArbovirus trasmessi da mosche (stimolo trasmessi da mosche (stimolo immunitario),immunitario),-- Euphorbia tirucalliEuphorbia tirucalli (danno al DNA),(danno al DNA),-- traslocazione coinvolgente traslocazione coinvolgente cc--MYCMYC..

LINFOMA DI BURKITTLINFOMA DI BURKITT

MORFOLOGIA:MORFOLOGIA: endemico/sporadico= isomorfiendemico/sporadico= isomorfitaglia media omogeneataglia media omogeneacromatina reticolarecromatina reticolare22--6 nucleoli6 nucleolicitoplasma basofilo/citoplasma basofilo/vacuolatovacuolatocrescita coesivacrescita coesivamitosi numerosissimemitosi numerosissimeaspetto a cielo stellatoaspetto a cielo stellato

LBT: dettagli citologiciLBT: dettagli citologiciin un caso con in un caso con colonizcoloniz--

zazionezazione ““follicolarefollicolare””

LINFOMA DI BURKITTLINFOMA DI BURKITT

FENOTIPO:FENOTIPO: SIgMSIgM++

marcatori Bmarcatori B++

CD10CD10++

BclBcl--66++

BclBcl--22--

cc--mycmyc++

MibMib--11++ = 100%= 100%

CD10CD10 BclBcl--66

BclBcl--22

KiKi--6767MIB1MIB1

GENOTIPOGENOTIPO

LBT endemico: LBT endemico: EBVEBV++

t(8;14)t(8;14)

LBT sporadico :LBT sporadico : EBV+: 25% HIVEBV+: 25% HIV--

50% HIV50% HIV++

t (2;8)t (2;8)t(8;22)t(8;22)diversi diversi breakpointsbreakpointsp53 (?)p53 (?)p21 (?)p21 (?)

Sul cromosoma 8: gene cSul cromosoma 8: gene c--MYCMYC

Cristina Campidelli, Lynnette Cristina Campidelli, Lynnette TumwineTumwine, Pier Paolo Piccaluga, , Pier Paolo Piccaluga, Stefano A PileriStefano A Pileri

BURKITT LYMPHOMA IN UGANDA IN THE THIRD MILLENNIUM: BURKITT LYMPHOMA IN UGANDA IN THE THIRD MILLENNIUM: A MORPHOLOGIC AND MOLECULAR APPRAISAL ON TISSUE A MORPHOLOGIC AND MOLECULAR APPRAISAL ON TISSUE

MICROMICRO--ARRAYARRAY

HIV infection and c-MYC status in endemic Burkitt lymphomaCampidelli, C., Gazzola, A., Vitone, F., Tumwine, L., and Pileri, S.A.Hum. Pathol., 39:1408-9, 2008.

Hum. Pathol., 39:817-823, 2008.

Patient and material collectionPatient and material collection•• 95 cases from Ugandan tribes95 cases from Ugandan tribes

•• 22--64 64 ysys (median age: 16)(median age: 16)

•• Site of involvement:Site of involvement:-- lymph node lymph node (34 cases)(34 cases)-- abdomen abdomen (26 cases)(26 cases)-- gonads gonads (25 cases)(25 cases)-- jaw jaw (10 cases)(10 cases)

•• formalin fixed paraffin embedded blocks punched formalin fixed paraffin embedded blocks punched and collected in 2 TMA blocks (48 and 47 cases and collected in 2 TMA blocks (48 and 47 cases each)each)

TMA technologyTMA technology

Results Results •• Positivity forPositivity for BB--cell markerscell markers, CD10 and Bcl, CD10 and Bcl--66

•• Negativity for BclNegativity for Bcl--22

•• KiKi--67 67 close toclose to 100%100%

•• ThirtyThirty--five CD30+ cases (37%)five CD30+ cases (37%)

•• Expression of CD38 in 73 cases (77%) and CD138 in 43 Expression of CD38 in 73 cases (77%) and CD138 in 43 cases (45%)cases (45%)

•• IRFIRF--4 mostly negative (7 positive cases)4 mostly negative (7 positive cases)

•• Constant EBV integrationConstant EBV integration

CD30CD30

CD138CD138

EBEREBER

HIVHIV--statusstatus

•• None of the samples showed HIV integrationNone of the samples showed HIV integration

•• HIVHIV--1 DNA was investigated by:1 DNA was investigated by:RTRT--PCR amplifying gag region (142 bp)PCR amplifying gag region (142 bp)nested PCR amplifying env region (248 bp)nested PCR amplifying env region (248 bp)

Driver GA et al. J Driver GA et al. J VirolVirol Methods, 2007Methods, 2007BrachtelBrachtel EF et al. AIDS Res Hum Retroviruses, 2002 EF et al. AIDS Res Hum Retroviruses, 2002 StrappeStrappe PM et al. J PM et al. J VirolVirol Methods, 1998Methods, 1998

FISH analysisFISH analysis•• FISH failed to demonstrate t(8;14) FISH failed to demonstrate t(8;14) due due to DNA degradationto DNA degradation

•• Surrogated by immunohistochemistrySurrogated by immunohistochemistry

•• Characteristic expression patterns of TCL1, CD38, and Characteristic expression patterns of TCL1, CD38, and CD44 identify aggressive lymphomas harboring a MYC CD44 identify aggressive lymphomas harboring a MYC translocation translocation ((RodigRodig SJ, Am J SJ, Am J SurgSurg PatholPathol, 2008) , 2008)

•• 64 out of 85 cases were TCL1+, CD38+ and CD4464 out of 85 cases were TCL1+, CD38+ and CD44--•• 9 out of 85 cases were TCL19 out of 85 cases were TCL1--, CD38+ and CD44, CD38+ and CD44--•• 9 out of 85 cases were TCL1+, CD389 out of 85 cases were TCL1+, CD38-- and CD44and CD44--•• 3 out of 85 cases were TCL13 out of 85 cases were TCL1--, CD38, CD38-- and CD44and CD44--

TCL1TCL1

CD38CD38

CD44CD44

QuestionQuestion

•• Are the three subtypes of Burkitt Are the three subtypes of Burkitt

lymphoma different diseases or different lymphoma different diseases or different

features of the same disease?features of the same disease?

Piccaluga PP, De Piccaluga PP, De FalcoFalco G, G, KustagiKustagi K, K, GazzolaGazzola A, A, AstolfiAstolfi A, Agostinelli C, A, Agostinelli C, LeucciLeucci E, E, OnnisOnnis A, A, TripodioTripodio C, C, SapienzaSapienza MR, MR, BellanBellan C, C, LazziLazzi S, S, TumwineTumwine L, L, MawandaMawanda M, M, OgwangOgwang M, M, CalbiCalbi V, Formica S, Califano A, Pileri V, Formica S, Califano A, Pileri SA and SA and LeonciniLeoncini L: Gene expression L: Gene expression analysis uncovers similarity and analysis uncovers similarity and differences among Burkitt lymphoma differences among Burkitt lymphoma subtypes. Blood 2011, Esubtypes. Blood 2011, E--pub ahead of pub ahead of print. print.

Burkitt lymphomaFollicular lymphomaDiffuse large B-cell lymphomaPrimary mediastinal B-cell lymphomaChronic lymphocytic leukemiaNormal B-cells

Molecular profiling of Molecular profiling of BL subtypesBL subtypes

BL is a unique molecular BL is a unique molecular entityentity

BL subtypes are all related to GC cells & presents BL subtypes are all related to GC cells & presents several deregulated genes and cellular programsseveral deregulated genes and cellular programs

SPARCSPARC BLBLGerminal Center B cellsGerminal Center B cells

+1+1 --11

NaNaïïveve

GerminalGerminal center Bcenter B --cellscells

MemoryMemory

BLBL

GreyGrey zonezone

GCGC N/MN/M

+1+1 --11

NaiveNaiveGerminalGerminal center B center B cellscellsMemoryMemoryBLBL

GreyGrey zonezone

GCGC N/MN/M

1,000 genes1,000 genes

•• immune response immune response •• proliferation (MYC)proliferation (MYC)•• adhesion adhesion •• NOTCH signalingNOTCH signaling

Though similar, BL subtypes present Though similar, BL subtypes present differences in their differences in their GEPsGEPs

eBL sBL HIV-BLeBLeBL sBLsBL HIVHIV-BLBL eBLHIV-BLsBL

eBLeBLHIVHIV --BLBLsBLsBL

eBLeBLsBLsBL

eBLeBLHIVHIV--BLBL

eBL and HIVeBL and HIV--BL: 16 genes BL: 16 genes (almost identical),(almost identical),

eBL & sBL: 254 genes, including eBL & sBL: 254 genes, including cell cycle regulation.cell cycle regulation.

Though similar, BL subtypes present Though similar, BL subtypes present differences in their differences in their GEPsGEPs

eBL (TAT & HIVeBL (TAT & HIV--BL)BL)

GSEA showed that eBL and sBL differ for the GSEA showed that eBL and sBL differ for the expression of genes related to the RBL2 network expression of genes related to the RBL2 network

RBL2 functional network identified byRBL2 functional network identified byARACNeARACNe

eBLeBLsBLsBL

The results do not vary byThe results do not vary bysubtracting the MYC networksubtracting the MYC network

Accordingly, RBL2 malfunction isAccordingly, RBL2 malfunction is relevant for eBL relevant for eBL but doesnbut doesn’’t affect sBL.t affect sBL.

ConclusionsConclusions1.1. Irrespective of the subtypes, BL is a unique molecular Irrespective of the subtypes, BL is a unique molecular

entity distinct from other Bentity distinct from other B--NHLs.NHLs.2.2. All BL subtypes are related to GC BAll BL subtypes are related to GC B--cells.cells.3.3. Genes and cellular programs deregulated in BL have Genes and cellular programs deregulated in BL have

been identified, including cell cycle and proliferation been identified, including cell cycle and proliferation control, NOTCH signaling and immune response.control, NOTCH signaling and immune response.

4.4. GEP differences do exist among BL subtypes.GEP differences do exist among BL subtypes.5.5. In particular, RBL2 malfunction seems to characterize In particular, RBL2 malfunction seems to characterize

the signature of eBL (and HIVthe signature of eBL (and HIV--BL?) suggesting a BL?) suggesting a possible involvement of such gene in BL pathogenesis.possible involvement of such gene in BL pathogenesis.