about b–cell lymphomas - computational diagnostics...
TRANSCRIPT
About B–Cell Lymphomas
Groupmeeting Klipp/Spang, December 09 2002
Dennis KostkaMax-Planck-Institute for Molecular Genetics
– Computational Molecular Biology –Berlin
1
Overview
• Short History of Lymphoma Classification
• About Healthy B–CellsBlood CellsImmune SystemImmunoglobulinsLymphatic SystemB–Cell Differentiation
Overview
• Short History of Lymphoma Classification
• About Healthy B–CellsBlood CellsImmune SystemImmunoglobulinsLymphatic SystemB–Cell Differentiation
• WHO Classification
Overview
• Short History of Lymphoma Classification
• About Healthy B–CellsBlood CellsImmune SystemImmunoglobulinsLymphatic SystemB–Cell Differentiation
• WHO Classification
• MicroarraysAlizadeh et. al.Shipp et. al.
2
Short History of Classification
• > 1970’sHenry Rappaport
Dennis Kostka About B-cell Lymphoma, MPI for Molecular Genetics, 2002 Dec 09 3
Short History of Classification
• > 1970’sHenry Rappaport
• 1970’sLukes, CollinsLennert, ’Kiel Group’
Dennis Kostka About B-cell Lymphoma, MPI for Molecular Genetics, 2002 Dec 09 3
Short History of Classification
• > 1970’sHenry Rappaport
• 1970’sLukes, CollinsLennert, ’Kiel Group’
• 1980’sWorking Formulation
Dennis Kostka About B-cell Lymphoma, MPI for Molecular Genetics, 2002 Dec 09 3
Short History of Classification
• > 1970’sHenry Rappaport
• 1970’sLukes, CollinsLennert, ’Kiel Group’
• 1980’sWorking Formulation
• 1994Revised European American Lymphoma Scheme
Dennis Kostka About B-cell Lymphoma, MPI for Molecular Genetics, 2002 Dec 09 3
Short History of Classification
• > 1970’sHenry Rappaport
• 1970’sLukes, CollinsLennert, ’Kiel Group’
• 1980’sWorking Formulation
• 1994Revised European American Lymphoma Scheme
• 2001WHO comprehensive classification of neoplasmsof haematopietic and lymphoid tissues
Dennis Kostka About B-cell Lymphoma, MPI for Molecular Genetics, 2002 Dec 09 3
Blood Cells
Blood Cells
Erytrocytes Leucocytes
Dennis Kostka About B-cell Lymphoma, MPI for Molecular Genetics, 2002 Dec 09 4
Blood Cells
Blood Cells
Erytrocytes Leucocytes
Lymphocytes Granulocytes Monocytes
Dennis Kostka About B-cell Lymphoma, MPI for Molecular Genetics, 2002 Dec 09 4
Blood Cells
Blood Cells
Erytrocytes Leucocytes
Lymphocytes Granulocytes Monocytes
T CellsB Cells NK Cells
Dennis Kostka About B-cell Lymphoma, MPI for Molecular Genetics, 2002 Dec 09 4
Immune System
Pathogens
Innate ImmuneResponses
Adaptive ImmuneResponses
• innate response
• adaptive response
Dennis Kostka About B-cell Lymphoma, MPI for Molecular Genetics, 2002 Dec 09 5
Immune System
Pathogens
Innate ImmuneResponses
Adaptive ImmuneResponses
• innate response
• adaptive responsecellular mediated
Dennis Kostka About B-cell Lymphoma, MPI for Molecular Genetics, 2002 Dec 09 5
Immune System
Pathogens
Innate ImmuneResponses
Adaptive ImmuneResponses
• innate response
• adaptive responsecellular mediatedantibody mediated
Dennis Kostka About B-cell Lymphoma, MPI for Molecular Genetics, 2002 Dec 09 5
Antibodies
how they work
how they look like
Dennis Kostka About B-cell Lymphoma, MPI for Molecular Genetics, 2002 Dec 09 6
The Lymphatic System
Human Lymphoid Organs
Lymphnode
Secondary Follicle
Dennis Kostka About B-cell Lymphoma, MPI for Molecular Genetics, 2002 Dec 09 7
Class Switch / Somatic Hypermutation
Class Switches
Somatic Hypermutation
• 1 / V region coding sequence / generation
• aim : enlargen affinity to antigen
• place: germinal center
Dennis Kostka About B-cell Lymphoma, MPI for Molecular Genetics, 2002 Dec 09 9
B Cell Differentiation
Dennis Kostka About B-cell Lymphoma, MPI for Molecular Genetics, 2002 Dec 09 10
B Cell Differentiation ctd.
Naive B cell
Dennis Kostka About B-cell Lymphoma, MPI for Molecular Genetics, 2002 Dec 09 11
B Cell Differentiation ctd.
Naive B cell
B Immunoblast
Dennis Kostka About B-cell Lymphoma, MPI for Molecular Genetics, 2002 Dec 09 11
B Cell Differentiation ctd.
Naive B cell
B Immunoblast Plasmacytoid Lymphocyte
Dennis Kostka About B-cell Lymphoma, MPI for Molecular Genetics, 2002 Dec 09 11
B Cell Differentiation ctd.
Naive B cell
B Immunoblast Plasmacytoid Lymphocyte
Mantle cell
Dennis Kostka About B-cell Lymphoma, MPI for Molecular Genetics, 2002 Dec 09 11
B Cell Differentiation ctd.
Naive B cell
B Immunoblast Plasmacytoid Lymphocyte
Mantle cell
Follicular B blast
Dennis Kostka About B-cell Lymphoma, MPI for Molecular Genetics, 2002 Dec 09 11
B Cell Differentiation ctd.
Naive B cell
B Immunoblast Plasmacytoid Lymphocyte
Mantle cell
Follicular B blast
Centroblast
Dennis Kostka About B-cell Lymphoma, MPI for Molecular Genetics, 2002 Dec 09 11
B Cell Differentiation ctd.
Naive B cell
B Immunoblast Plasmacytoid Lymphocyte
Mantle cell
Follicular B blast
Centroblast
Centrocyte
Dennis Kostka About B-cell Lymphoma, MPI for Molecular Genetics, 2002 Dec 09 11
B Cell Differentiation ctd.
Naive B cell
B Immunoblast Plasmacytoid Lymphocyte
Mantle cell
Follicular B blast
Centroblast
Centrocyte
Marginal zone & Monocytoid B cell
Dennis Kostka About B-cell Lymphoma, MPI for Molecular Genetics, 2002 Dec 09 11
B Cell Differentiation ctd.
Naive B cell
B Immunoblast Plasmacytoid Lymphocyte
Mantle cell
Follicular B blast
Centroblast
Centrocyte
Marginal zone & Monocytoid B cell
Dennis Kostka About B-cell Lymphoma, MPI for Molecular Genetics, 2002 Dec 09 11
B Cell Differentiation ctd.
Naive B cell
B Immunoblast Plasmacytoid Lymphocyte
Mantle cell
Follicular B blast
Centroblast
Centrocyte
Marginal zone & Monocytoid B cell
Plasma cell
Dennis Kostka About B-cell Lymphoma, MPI for Molecular Genetics, 2002 Dec 09 11
Blood Cancer
Blood Cancer
LeukemiaLymphoma Myeloma
CLL can beChronic Lymphatic LeukemiaSmall Lymphocytic Lymphoma
ALL can beAcute Lymphoblastic Leukemia (B-ALL)
if >25 % Lymphoblasts in bonemarrowLymphoblastic Lymphoma (B-LBL)
else
Dennis Kostka About B-cell Lymphoma, MPI for Molecular Genetics, 2002 Dec 09 12
Blood Cancer
Blood Cancer
LeukemiaLymphoma Myeloma
CLL can beChronic Lymphatic LeukemiaSmall Lymphocytic Lymphoma
ALL can beAcute Lymphoblastic Leukemia (B-ALL)
if >25 % Lymphoblasts in bonemarrowLymphoblastic Lymphoma (B-LBL)
else
each case: same biological entity
Dennis Kostka About B-cell Lymphoma, MPI for Molecular Genetics, 2002 Dec 09 12
WHO Classification
Haematological Malignancies
MyeloidLymphoid Histocytic / Dendritic
T Cells / NK Cells Hodkin Lymphomas
Precursor Mature
B Cells
Dennis Kostka About B-cell Lymphoma, MPI for Molecular Genetics, 2002 Dec 09 13
WHO Classification
Haematological Malignancies
MyeloidLymphoid Histocytic / Dendritic
T Cells / NK Cells Hodkin Lymphomas
Precursor Mature
ALL CLL
B Cells
Dennis Kostka About B-cell Lymphoma, MPI for Molecular Genetics, 2002 Dec 09 13
WHO Classification
Haematological Malignancies
MyeloidLymphoid Histocytic / Dendritic
T Cells / NK Cells Hodkin Lymphomas
Precursor Mature
ALL CLL
B Cells
85%15%
30%
Dennis Kostka About B-cell Lymphoma, MPI for Molecular Genetics, 2002 Dec 09 13
WHO Classification ctd.
Criteria:
• Clinical Presentation· dissaminated?· leuekemic?· extranodal/nodal?
• Morpholgy· ’structure’ of cells involved
• Immunophenotype· CD surface markers, Immunoglobulins
• Cytogenetics· translocations
Dennis Kostka About B-cell Lymphoma, MPI for Molecular Genetics, 2002 Dec 09 14
Translocations
• deregulate oncogene by placing it e. g. near IG locioncogenes have been found that way
• create new fusion proteins
Dennis Kostka About B-cell Lymphoma, MPI for Molecular Genetics, 2002 Dec 09 15
WHO B–Cell Neoplasms
I. Precursor B-cell neoplasm:A. precursor B-acute lymphoblastic leukemia/lymphoblastic lymphoma (B-ALL, LBL)
II. Peripheral B-cell neoplasmsA. B-cell chronic lymphocytic leukemia/small lymphocytic lymphomaB. B-cell prolymphocytic leukemiaC. Lymphoplasmacytic lymphoma/immunocytomaD. Mantle cell lymphomaE. Follicular lymphoma (22%)F. Extranodal marginal zone B-cell lymphoma of MALT typeG. Nodal marginal zone B-cell lymphoma (+/- monocytoid B-cells)H. Splenic marginal zone lymphoma (+/- villous lymphocytes)I. Hairy cell leukemiaJ. Plasmacytoma/plasma cell myelomaK. Diffuse large B-cell lymphoma (30%)L. Burkitt’s lymphoma
(subtypes are missing)
Dennis Kostka About B-cell Lymphoma, MPI for Molecular Genetics, 2002 Dec 09 16
Diffuse Large B–Cell Lymphoma
Dennis Kostka About B-cell Lymphoma, MPI for Molecular Genetics, 2002 Dec 09 17
Diffuse Large B–Cell Lymphoma
• Account for about 30% of B–cell neoplasms
Dennis Kostka About B-cell Lymphoma, MPI for Molecular Genetics, 2002 Dec 09 17
Diffuse Large B–Cell Lymphoma
• Account for about 30% of B–cell neoplasms
• Two variants (morphological)· Immunoblastic· Centroblastic
• Highly variable clinical course (independent of subtype)
Dennis Kostka About B-cell Lymphoma, MPI for Molecular Genetics, 2002 Dec 09 17
Diffuse Large B–Cell Lymphoma
• Account for about 30% of B–cell neoplasms
• Two variants (morphological)· Immunoblastic· Centroblastic
• Highly variable clinical course (independent of subtype)
use microarrays for further classification
Dennis Kostka About B-cell Lymphoma, MPI for Molecular Genetics, 2002 Dec 09 17
Microarrays: Alizadeh et al.
Lymphochip· 17 856 cDNA clones· 96 tissue-samples· DLBCL, FL, CLL· 128 chips
Alizadeh, A. et.al Distinct types of dif-fuse large B-cell lymphoma identified bygene expression profiling. Nature 4051,503-511 (2000)
OCI Ly3OCI Ly10DLCL-0042DLCL-0007DLCL-0031DLCL-0036DLCL-0030DLCL-0004DLCL-0029Tonsil Germinal Center BTonsil Germinal Center CentroblastsSUDHL6DLCL-0008DLCL-0052DLCL-0034DLCL-0051DLCL-0011DLCL-0032DLCL-0006DLCL-0049TonsilDLCL-0039Lymph NodeDLCL-0001DLCL-0018DLCL-0037DLCL-0010DLCL-0015DLCL-0026DLCL-0005DLCL-0023DLCL-0027DLCL-0024DLCL-0013DLCL-0002DLCL-0016DLCL-0020DLCL-0003DLCL-0014DLCL-0048DLCL-0033DLCL-0025DLCL-0040DLCL-0017DLCL-0028DLCL-0012DLCL-0021Blood B;anti-IgM+CD40L low 48hBlood B;anti-IgM+CD40L high 48hBlood B;anti-IgM+CD40L 24hBlood B;anti-IgM 24hBlood B;anti-IgM+IL-4 24hBlood B;anti-IgM+CD40L+IL-4 24hBlood B;anti-IgM+IL-4 6hBlood B;anti-IgM 6hBlood B;anti-IgM+CD40L 6hBlood B;anti-IgM+CD40L+IL-4 6hBlood T;Adult CD4+ Unstim.Blood T;Adult CD4+ I+P Stim.Cord Blood T;CD4+ I+P Stim.Blood T;Neonatal CD4+ Unstim.Thymic T;Fetal CD4+ Unstim.Thymic T;Fetal CD4+ I+P Stim.OCI Ly1WSU1JurkatU937OCI Ly12OCI Ly13.2SUDHL5DLCL-0041FL-9FL-9;CD19+FL-12;CD19+FL-10;CD19+FL-10FL-11FL-11;CD19+FL-6;CD19+FL-5;CD19+Blood B;memoryBlood B;naiveBlood BCord Blood BCLL-60CLL-68CLL-9CLL-14CLL-51CLL-65CLL-71#2CLL-71#1CLL-13CLL-39CLL-52DLCL-0009
2 1 0–1–2
4.0002.0001.0000.5000.250
DLBCLGerminal centre BNl. lymph node/tonsilActivated blood BResting/activated TTransformed cell linesFLResting blood BCLL
Germinal CentreB cell
Lymph node
T cell
Pan B cell
Activated B cell
Proliferation
A
G
Dennis Kostka About B-cell Lymphoma, MPI for Molecular Genetics, 2002 Dec 09 18
Alizadeh ctd.
a b c
GC B-like DLBCL Activated B-like DLBCL
DLC
L-00
12D
LCL-
0024
DLC
L-00
03D
LCL-
0026
DLC
L-00
23D
LCL-
0015
DLC
L-00
10D
LCL-
0030
DLC
L-00
34S
UD
HL6
DLC
L-00
18D
LCL-
0032
DLC
L-00
52D
LCL-
0037
DLC
L-00
01D
LCL-
0008
GC
BG
C c
entr
obla
sts
DLC
L-00
04D
LCL-
0029
DLC
L-00
09D
LCL-
0020
DLC
L-00
51D
LCL-
0033
DLC
L-00
05D
LCL-
0011
DLC
L-00
48D
LCL-
0027
DLC
L-00
13D
LCL-
0007
DLC
L-00
28D
LCL-
0025
DLC
L-00
21D
LCL-
0016
DLC
L-00
02D
LCL-
0017
OC
I Ly3
DLC
L-00
40D
LCL-
0014
DLC
L-00
31D
LCL-
0036
DLC
L-00
39D
LCL-
0042
OC
I Ly1
0D
LCL-
0041
DLC
L-00
49D
LCL-
0006
10 12
1.0
0.5
0.00 2 4 6 8
G C B -like
Activated B -likePro
bab
ility
19 patients , 6 deaths
21 patients , 16 deaths
P =0.01
12
1.0
0.5
0.00 2 4 6 8 10
Pro
bab
ility
P =0.002
Overall survival (years )
Low clinical risk
24 patients , 9 deaths
High clinical risk
14 patients , 11 deaths
All patients
All patients
a
b
c
12
1.0
0.5
0.0
0 2 4 6 8 10
Pro
bab
ility
P =0.05
G C B -like
14 patients , 3 deaths
Activated B -like
10 patients , 6 deaths
Low clinical risk patients
Dennis Kostka About B-cell Lymphoma, MPI for Molecular Genetics, 2002 Dec 09 19
Microarrays: Shipp et al.
Affy HU 6800· 6 800 genes· 77 tissue samples· DLBCL, FL
Shipp, M.A. et al. Diffuse large B-celllymphoma outcome predition by geneexpression profiling using supervisedmachine learning. Nature Medicine 8,68-74 (2002)
Dennis Kostka About B-cell Lymphoma, MPI for Molecular Genetics, 2002 Dec 09 20