Dysplasie in AML en (cyto)genetische afwijkingen: Chromosoom

 Dysplasie in AML en (cyto)genetische
afwijkingen:
 Chromosoom 3q26 afwijkingen en EVI1
patient 0390606
BM
H11-172
8-2-2011
patient 0390606
dag 24 kuur I H11-428
4-4-2011
patient 0390606
dag 24 kuur I H11-428
4-4-2011
 Het betreft een patiente uit 1958.
bij presentatie Hb 6.8
WBC 5.6
thrombo 412.
Diagnose WHO 2008: AML met t(3;3)(q21;q26)RPN1-EVI1
behandeld in de HOVON 42:
na 1e kuur geen response
na 2e kuur morfologisch en immunologisch in remissie
daarna allo transplantatie
en na 3 weken recidief en overleden.
Cytogenetica: t(3;3) en -7.
Acute myeloid leukemia: Molecular biology
Normal
AML
Bone marrow: Morphology
Normal
Leukemia
Normal bloodcell formation
Stemcell
Bone marrow
Blood
One cell transforms:
The beginning of leukemia.
Bonemarrow
Immature
cell
Blood
Leukemia cells in bone marrow and blood
Bone marrow
Blood
AML behaves differently in different patients
patient A
† in 1 month
patient B
cured
patient C
Cured, but one year later †
 Why do AML patients respond differently to
therapy?
 Why do AML patients respond differently to
therapy?
• AML is not one disease
• AML is a disease of the genes
• Different AMLs have different recurrent genetic
abnormalities
Identification of genetic defects in AML:
• Improved diagnostics and prognostics
• More specific treatment protocols
Identification of genetic defects in AML:
• Improved diagnostics and prognostics
• More specific treatment protocols
Normal: 2 x 23 chromosomes
Kern
Cel
DNA
AML is a clonal disease with frequent
chromosomal abnormalities.
Translocation t(8;21): an example
Acute myeloid leukemia
Prognosis based on chromosomal abnormalities
percentage living
100
inv(16)
t(15;17)
t(8;21)
favorable
50
intermediate
unfavorable
-7/7q-, t(3;3)/inv3, t(6;9), t(9;22), complex, a.o.
0
0
1
2
years
3
4
5
Chromosome analysis
Favorable
treatment
t(8;21)
Chemotherapy
Intermediate
poor
Inv (3)
Chemotherapy
+
Transplantation
Chromosome analysis
Good
treatment
t(8;21)
Chemotherapy
Intermediate
poor
Inv (3)
?
Chemotherapy
+
Transplantation
Chromosome 3q26 aberrations in AML
3q26 aberrations: ~2% AML
Inv 3
t(3;3)
Chromosome 3q26 aberrations in AML
3q26 aberrations: ~2% AML
1.2
AML with 3q26 aberrations
Inv 3
t(3;3)
Cum Survival
1.0
AML intermediate risk
.8
.6
.4
.2
0.0
0
20
40
60
80
100
120
Event free survival, [m]
140
160
Chromosome 3q26 aberrations in AML
3q26 aberrations: ~2% AML
Inv 3
t(3;3)
3
3
3
3
EVI1 is the gene that is located at the breakpoint
region at Chromosome 3q26
3q26 aberrations: ~2% AML
3
Inv 3
Evi1
t(3;3)
3
Identification of genetic defects and analysis
gene expression in AML:
INTERMEZZO I
Two cells from the same
individual and thus with the
same DNA
23 chromosomes: ~20.000 genes.
Gene: turned on
Gene: turned off
DNA
Normal myeloid cell
Skin cell
Liver cell
22 paar chromosomen
XX of XY
22.000 genen/cel
Een chromosoom
DNA
DNA
DNA
Genen coderen voor eiwitten
22.000 genen/cel
Een Gen
start
stop
DNA
RNA polymerase
ATG
TGA
AAAA
RNA
ribosomen
M-AZ-AZ-AZ-AZ-AZ-AZ
EIWIT
EVI1 is the gene that is located at the breakpoint
region at Chromosome 3q26
3q26 aberrations: ~2% AML
3
Inv 3
Evi1
t(3;3)
3
The EVI1 gene is located on Chromosome 3 is switched off in
normal bone marrow cells.
Normal
marrow
EVI1 off
Chromosome 3
The EVI1 gene located on Chromosome 3 is switched on in AML
with a chromosome 3q26 break.
Normal
marrow
EVI1 off
Chromosome 3
Leukemia
EVI1 on
Chromosome 3:
break
treatment
Favorable
t(8;21)
Chemotherapy
Intermediate
?
EVI1
?
Poor
Inv (3)
Chemotherapy
= EVI1
+
Transplantation
The EVI1 gene is turned on in ~10 % subset of AML patients without
a chromosome 3 break.
Normal
marrow
EVI1 off
Chromosome 3
Leukemia
EVI1 on
Chromosome 3:
break
Leukemie
EVI1 on
Chromosom3
Treatment
Favorable
Intermediate
EVI1 expression without
3q26 break(`~10%)
?
Poor
+
Combined AML cohorts revealed prognostic impact
of high EVI1 levels
Overall Survival (%)
Relapse-free Survival (%)
Event-free Survival (%)
100
100
100
75
75
75
50
50
50
EVI1- n=1234
EVI1- n=952
EVI1- n=1234
25
25
EVI1+ n=148
25
EVI1+ n=79
EVI1+ n=148
P < .001
0
0
2
4
6
8
10
12
14
16
18
Time (years)
P < .0001
0
0
2
4
6
8
10
12
14
16 18
P < .001
0
0
2
4
6
Time (years)
8
10
12
14
16
Time (years)
Achievement of CR
HR, 0.54; 95%CI, (0.36-0.80), P=0.002
OS
HR, 1.17; 95%CI, (0.93-1.46), P=0.18
EFS
HR,1.46; 95%CI, (1.19-1.87), P=0.0003
DFS
HR, 1.32 ; 95%CI, (0.99-1.76),P=0.05
*Cox regression model stratified for cohorts and other prognostic AML markers, age, WBC, platelets, cytogenetic risk, type of AML and NPM1wt/FLTITDneg.
EVI1+ AML cases benefit of allogeneic
transplant in first CR
A
B
100
75
Allo-TPL in 1st CR n=28
50
No Allo-TPL in 1st CR n=51
25
P = .05
0
0
1
2
3
4
5
6
7
8
Time (years)
9
10 11 12
Relapse-free Survival (%)
Overall Survival (%)
100
75
50
Allo-TPL in 1st CR n=28
25
No Allo-TPL in 1st CR n=51
P = .001
0
0
1
2
3
4
5
6
7
8
Time (years)
9
10 11 12
Favorable
Intermediate
EVI1 expression without
3q26 break (10%)
+
Poor
+
De search for other genetic abnormalities continues.
Favorable
Intermediate
Other mutation?
Poor
+
Gene expression profiling in AML
23 chromosomes: ~20.000 genes.
Gene: turned on
Gene: turned off
DNA
Leukemia A
Leukemia B
Leukemia C
Gene expression profiling of AML:
Expression analysis of 13.000 genes
The Genetic (bar)code
Gen expressie profiel analyse bij AML:
Expressie analyse van 13.000 genen
The Genetic (bar)code
AMLs zijn morphologisch vergelijkbaar:
Een studie bij 285 patienten.
…
AML 1
AML 2
AML 3
AML 4
AML 5
…………………………………..
…………………………………..
………….
AML 283
AML 284 AML 285
Gen expressie profielen bij AML:
Opslaan van de genetische codes bij 285 AML
patiënten
…
AML 1
AML 2
AML 3
AML 4
AML 5
…………………………………..
…………………………………..
………….
AML 283
AML 284 AML 285
Gene expression profiling
Omniviz
Pearson's correlatie coefficient analyse
B
A
Omniviz
Pearson's correlatie coefficient analyse
A
B
A
D
C
A
E
A
A
F
A
A
Analyse en vergelijking van de gen expressie profilen
bij 6 AML patiënten
285 AML patients (random order)
1
2
3
4
5
6
1
2
3
4
5
6
285 AML patients
Analyse en vergelijking van de gen expressie profilen
bij 50AML patiënten
50 AML patienten
285 AML patients
50 AML patienten
285 AML patients (random order)
Analyse en vergelijking van de gen expressie profielen
285 AML patienten
285 AML patients
285 AML patienten
285 AML patients (random order)
Clustering van AML patiënten met “zelfde”
gen expressie profiel
285 AML patients
285 AML patienten
285 AML patients
285 AML patienten
285 AML patienten kunnen worden onderverdeeld
in 16 subgroepen of clusters
285 AML patienten
285 AML patienten kunnen worden onderverdeeld
in 16 subgroepen of clusters
285 AML patienten kunnen worden onderverdeeld
in 16 subgroepen of clusters
Patienten met dezelfde bekende translocaties zitten in een
zelfde cluster: Zelfde gen expressie profiel
Inv(16)
t(15;17)
t(8;21)
Wat betekent dit voor de andere clusters?
Inv(16)
?????
t(15;17)
t(8;21)
Chromosome 3q26 defects in AML cases from cluster
10:
EVI1 defects
EVI1 +/EVI1
-
AML cases from cluster 10 respond poor to treatment
Event free survival
Cumulative percentage
100
75
t(8;21), inv 16, t(15;17)
50
25
Cluster 10
0
0
Months
60
Gene profiling of AML
• Identification of genetically defined AML
• Subgroups within genetically defined AML
• Novel subclasses of AML
• Novel insight into the biology of AML
Acknowledgements
•
Erasmus MC
•
University of Ulm
•
•
•
•
•
•
•
•
•
•
•
Sanne Lugthart
Marije Havermans
Bob Löwenberg
Claudia Erpelinck
Eric Bindels
Chantal Goudswaard
Berna Beverloo
Kirsten van Lom
Antoinette van Hoven-Beijen
Roel Verhaak
Peter Valk
•
•
•
•
•
Stefan Gröschel
Richard F. Schlenk
Karina Eiwen
Hartmut Döhner
Konstanze Döhner