Linfoma de Hodgkin - Comunidad Virtual de Anatomía ... · Hodgkin’s Lymphoma Two diseases Nodular lymphocyte predominant Hodgkin’s Lymphoma Classical Hodgkin’s Lymphoma •Nodular

Linfoma de

Hodgkin

Juan F García

Hodgkin’s Hodgkin’s LymphomaLymphoma

Two diseasesTwo diseases

Nodular lymphocyte predominant Hodgkin’s Lymphoma

Classical Hodgkin’s Lymphoma•Nodular sclerosis HL•Lymphocyte-rich classical HL•Mixed cellularity HL•Lymphocyte depletion HL

NLPHLNLPHL

• Origen en células B del CG

Rasgos clínicos• Niños y adultos jóvenes• Estadios localizados• Curso clínico indolente• Puede progresar a linfoma difuso de

células grandes

Patología• Patrón nodular• Células tumorales de tipo L&H, con

fenotipo semejante a células B del CG (CD20+, CD79a+, Bcl6+, OCT2+, PAX5+, Ig+, Bcl2-,...)

• EMA+, CD30-, CD15-• Background: linfocitos B maduros.

Células T CD57+• No asociación con EBV

Formas clásicas de LHFormas clásicas de LH

• Origen en células B del CG

Rasgos clínicos• Edad variable. Picos de incidencia en la

2ª y 5ª década• Puede presentarse en estadios

avanzados• Linfoma agresivo, de curso fatal sin

tratamiento• Rara progresiónPatología• Patrón variable según tipo histológico• Células tumorales de tipo H&RS, con

ausencia de marcadores propios de células B

• CD30+, CD15+ • Background: polimorfo: células T,

neutrófilos, eosinófilos,histiocitos, células plasmáticas,...

• EBV 40-70%

Nodular lymphocyte predominant Nodular lymphocyte predominant Hodgkin’s Lymphoma Hodgkin’s Lymphoma (NLPHL)(NLPHL)Nodular ParagranulomaNodular Paragranuloma

Rasgos clínicos

• Niños y adultos jóvenes, estadios localizados, curso clínico indolente

• ...pero puede progresar a linfoma difuso de células grandes (confrecuencia morfología tipo TCRBCL)

Morfología y fenotipo

• Patrón nodular: folículos irregulares con una reacción anormal del centro germinal (transformación progresiva)

• Células tumorales de tipo L&H, con fenotipo semejante a células B del CG: CD20+, CD79a+, Bcl6+, OCT2+, PAX5+, MUM1-/+, Ig+, Bcl2-

• EMA+, CD30-, CD15-

• Background: nódulos de linfocitos B maduros. Histiocitos. Células T CD57+

• No asociación con EBV

NLPHLNLPHL

NLPHLNLPHL

IgDIgD

CD20CD20 BCL2BCL2

CD57CD57OCT2OCT2

NLPHLNLPHL

NLPHLNLPHL

L&H L&H cellscells

BCL6BCL6EMAEMA

CD20CD20CD20CD20

OCT2OCT2

CD57+ TCD57+ T--cellscells

NLPHL: pathology

• Nodular +/- diffuse pattern• Neoplastic cells: L&H cells

• CD20+, CD79a+, BCL6+, BCL2-, OCT2+, PAX5+, MUM1-/+, EMA+, CD30-, CD15-

• Background:• B-cell nodules (PTGC), CD20+, IgD+, BCL2+, BCL6-,

CD10-• CD57+ T-cells (rosettes).• Histiocytes• Variable fibrosis

Classical Hodgkin’s Lymphoma

Def.: primary lymphoid malignancy in which tumour cells (H&RS cells) usually represent a minor population (<10%) within the affected tissue, which mainly consists of a heterogeneous infiltrate of inflammatory cells.

• Origin: GC B-lymphocytes• Phenotype:

• CD20-/+, CD79a-, Bcl6-, Igs-, OCT2-, PAX5+, MUM1+, Bcl2+ (30-40%)• CD30+, CD15+ (70-80%), EBV(LMP1)+ (40-70%)

• Genetics: • Rearranged and somatically mutated IgH genes• Gains of 9p and 2p

CD30CD30> 90%> 90%

CD15CD157070--80%80%

CD20CD20

CD79aCD79a

PAX5PAX5

BCL2BCL2

Bcl2Bcl2

Months

140120100806040200

Ove

rall S

urvival (Bcl2

)

1,1

1,0

,9

,8

,7

,6

,5

,4

,3

,2

,1

0,0

Bcl2 +

Rassidakis GZ, et al. BCL-2 expression in Hodgkin and Reed-Sternberg cells of classical Hodgkin disease predicts a poorerprognosis in patients treated with ABVD or equivalent regimens. Blood 2002;100(12):3935-41

Garcia JF, et al. Hodgkin and Reed-Sternberg cells harbor alterations in the major tumor suppressor pathways and cell-cycle checkpoints: analyses using tissue microarrays. Blood 2003;101(2):681-9

Brink AA, et al. Low p53 and high bcl-2 expression in Reed-Sternberg cells predicts poor clinical outcome for Hodgkin's disease: involvement of apoptosis resistance? Mod Pathol. 1998;11:376-383

Classical HL: pathology• Variable histological features:

subtypes NS, MC, LRCHL, LD• Neoplastic cells: H&RS cells, and

variants: mononuclear (Hodgkin’s) cells, lacunar cells, mummy cells.

<5%• Ausente o

fibrosis difusa

• Escaso• Numerosa y pleomórfica

DL

<5%• Puede

existir• Nódulos de

linfocitos B maduros

• Histiocitos

• Escasas células de H/RS, mononucleares

LHCRL

20-40%

50-70%

%

• Usualmente ausente

• Heterogéneo, histiocitos, eosinófilos y células plasmáticas

• Numerosas célulasde H/RS y variantes

• Células mumificadas

CM

• Abundante• Gruesos

septos que dividen nódulos

• Abundantes eosinófilos

• Numerosas células lacunares

• Variable número de células de H/RS

EN

FibrosisFondo reactivo

Celularidad neoplásicaSubtipo

Nodular Sclerosis HL

• Young females

• Mediastinum

• Usually EBV-

NS HLNS HL

H&RS + H&RS + lacunar cellslacunar cellsCD30CD30

Mixed Cellularity HL

• Males>Females

• Localized or disseminated disease

• More frequent EBV+

• Common in HIV+ patients

MC HLMC HL

MC HLMC HL

MummyMummy cellcell

EBV LMP1EBV LMP1

““InterfollicularInterfollicular” HL & ” HL & CastlemanCastleman--like featureslike features

CD30CD30

Lymphocyte Rich Classical HL

• Rare

• Usually stages I&II disease

• Less aggressive

• EBV-/+

• Differential diagnosis with NLPHL

LRCHL

LRCHL CD20

CD30

EBVLMP1

Lymphocyte Depleted HL

• Rare

• Older patients, advanced stages

• Poor prognosis

• Some cases in HIV+ patients show LD morphology

LD HL

Pathology

+/-

Broad septa in NSAbsent or Variable in MC,

LR, and LD

Fibrosis

NLPHL

Classical HL

• B Lymphocytes• CD57+ T-cells

L&H

Polymorphous:• Neutrophils• Eosinophils• Plasma cells• T-cells

H&RSVariants: lacunarcells (NS), mummy cells (MC)

BackgroundNeoplastic cells

Phenotype

NLPHL

Classical HL

+

-/+

EMA

-

30-40%

EBV (LMP)

-

+/-

CD15

-

+

CD30

+

-/+

OCT2

+

+

PAX5

-/+++

+--/+

MUM1CD79aCD20

+/-

-/+

BCL2

+

-

BCL6

Phenotype

DLBCL

HL

10-15%LMP1-

30-70%LMP1+

EBV

-

+/-

CD15

-+

+

CD30

+

-/+

OCT2

+

+

PAX5

-/+++

+--/+

MUM1CD79aCD20

Differential diagnosis between classical HL and DLBCL

Morphology !!!Tumoral cells & reactive background

Large B-Cell Lymphoma with Hodgkin’s Features

CD30

CD20

OCT2

PAX5

CD79aCD15


• ‘Grey zone’ lymphoma: neoplasms with morphological and immunophenotypic features intermediate between DLBCL and HL

• Usually young males with mediastinalinvolvement, diagnosed at advanced stages

• Tumors with similar morphology and immunophenotype may also present in extramediastinal locations.


Patofisiología del Linfoma de Hodgkin

•• Origen celular: Origen celular: •• linfocitos B del centro germinallinfocitos B del centro germinal

Cuestiones abiertas sobre el LHCuestiones abiertas sobre el LHY la bY la biologiologííaa de la cde la céélula de H&RSlula de H&RS

Rearranged and somatically mutated IgH genes in tumoral cells of classical HL and NLPHL

Bräuninger et al, 1999a; Irsch et al, 1998; Kanzler etal, 1996a, 1996b; Küppers et al, 1994; Marafioti et al,1999, 2000; Müschen et al, 2000; Ohno et al, 1998;Vockerodt et al, 1998,....


•• SSííntesis defectiva de inmunoglobulinasntesis defectiva de inmunoglobulinas•• DDééficit de factores de transcripcificit de factores de transcripcióón (OCT2, OCT1, BOB1)n (OCT2, OCT1, BOB1)•• Mutaciones Mutaciones ““cripplingcrippling”” de los genes de las de los genes de las IgsIgs


CÉLULA B NAIVE

CÉLULA BPROGENITORA

IgM+/D+

MANTO

IGM+/D+

SELECCIÓNANTIGÉNICA

SIg+

IgM+/D-

CÉLULA B DE MEMORIA

IgG/A+

CÉLULA PLASMÁTICA

IgG/A+

HIPERMUTACIÓNSOMÁTICA

CENTRO GERMINAL

Apoptosis

Selecciónnegativa

IgM+/D-

SIg-



•• Perdida de identidad linfoide BPerdida de identidad linfoide B


Gene expression profile ofHodgkin’s Lymphoma cell lines

5 well characterised HL-derived cell lines:L540L428

HDLM2KMH2L1236

Two independent cultures from each line

Normal germinal centre B-cells (CD77+, centroblasts) isolated from reactive tonsils.

L12

36L

428

HD

LM

2L

540

KM

H2

CD

77+

Gene expression profile of Hodgkin’s Lymphoma cell lines:Inactivation of B-cell receptor signalling and B-cell differentiation program

BCL6 FOS

IRF4

B-CELL RECEPTOR

CD19

CD20

CD22

CD79ACD79B

CD10

MHC class II

CD40CD40

BRDG1

HCLS1

TNFRSF17(B-cell maturation factor)

LYN

SYK

PKC

VAV

BTK



•• Perdida de identidad linfoide BPerdida de identidad linfoide B•• Diferentes mecanismos de supervivenciaDiferentes mecanismos de supervivencia

•• ActivaciActivacióón de NFn de NF--kappaB kappaB (CD30, EBV, JAK2, (CD30, EBV, JAK2, STATsSTATs, , IkBaIkBa,...),...)•• Programa B defectivoPrograma B defectivo•• DesregulaciDesregulacióón de los sistemas de control del ciclo celular y de n de los sistemas de control del ciclo celular y de

la apoptosis la apoptosis


Identification of a gene expression signature associated with unfavourable treatment

response in Hodgkin’s Lymphoma

Abel Sánchez-AguileraLymphoma GroupMolecular Pathology ProgrammeSpanish National Cancer Centre (CNIO)

Gene expression profile of Hodgkin’s Lymphoma

Tissue samples from HL patients with well-defined treatment response (29 samples)

Stringent selection criteria:• Classical HL (nodular sclerosis or mixed cellularity), HIV-, • Advanced stages (IIB, III, IV).• Treated with standard chemotherapeutic protocols (ABVD or variants, containing adriamycin).• Availability of frozen tissue from pre-treatment biopsy.• Follow-up of at least 2 years.• Good treatment response (14) = sustained complete remission.• Poor treatment response (15) = no complete remission, or relapse within 12 months after treatment.

Fluorescent labelling and hybridisation vs Universal Reference RNA (Stratagene)CNIO OncoChip v2 (13471 human clones)

RNA amplification by T7 in vitro transcription

RNA extraction

Control samples:• 5 reactive lymph node samples.• 5 HL-derived cell lines.• Normal centroblasts.

Hierarchical clustering of genes associated with treatment response

CLUSTER 1Immune response

CLUSTER 2Extracellular matrixAdhesionCell-cell signalling

CLUSTER 3ApoptosisSignal transduction

CLUSTER 4Cell cycle

CD8B1CD3DSH2D1AITM2A

ALDH1A1LYZSTAT1

MAD2L1CDC2CHEK1STK6

TOP2APCNARRM2TYMS

CYCSCASP14PDCD10STK17A

PRKACBPPP1CARAB27ACDH1

FZD4 CR1CCL26HLA-DRB3

CELL LINES

Centrob

lasts

Lymph

nodes

Goodresponse Poorresponse

HODGKIN’S LYMPHOMAS

Rel.

PTGS1HSPG2TIMP4

Role of T and NK-cells

Alvaro T, et al. Outcome in Hodgkin's lymphoma can be predicted from the presence of accompanying cytotoxic and regulatory T cells. Clin Cancer Res 2005. 11(4):1467-73.

Comparison of survival time (OS, EFS, DFS) according to TIA-1+ cells and FoxP3+:

(a) tumor infiltrate with a low level of TIA-1+ cells and a high level of FoxP3+ cells,

(b) tumor infiltrate with a high level of TIA-1+ cells and a low level of FoxP3+ cells,

(c) tumor infiltrate with intermediate levels of TIA-1+ and FoxP3+ cells.

Role of T and NK-cells

Alvaro T, et al. Clin Cancer Res 2005. 11(4):1467-73.

Hierarchical clustering of genes associated with treatment response

CLUSTER 1Immune response

CLUSTER 2Extracellular matrixAdhesionCell-cell signalling

CLUSTER 3ApoptosisSignal transduction

CLUSTER 4Cell cycle

CD8B1CD3DSH2D1AITM2A

ALDH1A1LYZSTAT1

MAD2L1CDC2CHEK1STK6

TOP2APCNARRM2TYMS

CYCSCASP14PDCD10STK17A

PRKACBPPP1CARAB27ACDH1

FZD4 CR1CCL26HLA-DRB3

CELL LINES

Centrob

lasts

Lymph

nodes

Goodresponse Poorresponse

HODGKIN’S LYMPHOMAS

Rel.

PTGS1HSPG2TIMP4

Identification of genes involved in HL progression:

Genes related with genome integrity checkpoints and G2/M checkpoint

Analysis of centrosomes and spindle checkpoint in H&RS cell lines: disrupted transition through mitosis in Hodgkin cells

hyper-G2/M

0

5

10

15

20

25

30

PBL L1236 HDLM2 L428

apoptosis

0

10

20

30

40

50

60

70

PBL L1236 HDLM2 L428

untreated

20 h

48 h

untreated

20 h

48 h

Validation of the expression data in an independentseries of patients.

235 cases IHC on TMAs

Linfoma de Hodgkin

Dr. Juan F García

Documents

Linfoma de Hodgkin - Comunidad Virtual de Anatomía ... · Hodgkin’s Lymphoma Two diseases Nodular lymphocyte predominant Hodgkin’s Lymphoma Classical Hodgkin’s Lymphoma •Nodular