JOURNAL TRANSCRIPT
Aneuploidy in Leukemia and Cancer Claudia Haferlach MLL Munich Leukemia Laboratory
Cancer is a genetic disease Activation of oncogenes
Inactivation of tumorsuppressor genes
Formation of tumorspecific fusion genes
t(8;14)(q24;q32) -> CMYC overexpression amplification of MLL
17p deletion + mutation of the 2nd TP53 allele
t(15;17)(q22;q12) -> PML-RARA
uncontrolled proliferation blocked differentiation apoptosis resistance
Cancer
Definition •
Aneuploidy –
loss and/or gain of whole chromosomes
Trisomy 8
Monosomy 7
• Aneuploidy is found in most solid tumors and half of all leukemias and lymphomas (http://cgap.nci.nih.gov/Chromosomes/Mitelman)
How does aneuploidy occur?
Thompson S. et al. Chromosome Res 2011
Many „players“ protect against aneuploidy
Holland A et al., Nat Rev Mol Cell Biol 2011
Pathways to the generation of aneuploidy
Holland A et al., Nat Rev Mol Cell Biol 2011
Gain of chromosomal material
46,XX
47,XX,+8
ALL with high hyperdiploidy
ALL with low hypodiploidy
ALL with near triploidy
Moorman A et al., Blood 2007
Number of unbalanced aberrations 45% 40% 35% 30% 25% 20% 15% 10% 5% 0% 1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19 ≥ 20
MLL data
Number of unbalanced aberrations 45% 40% 35% 30% 25% 20% 15% 10% 5% 0% 1
2
3
4
5
ALL (n=241)
6
7
8
9
10
11
12
13
14
15
16
17
18
19 ≥20
ALL
MLL data
Number of unbalanced aberrations 45% 40% 35% 30% 25% 20% 15% 10% 5% 0% 1
2
3
4
5
ALL (n=241)
6
7
8
9
10
11
12
13
14
15
16
17
18
19
≥20
ALL CLL CLL (n=3046)
MLL data
Number of unbalanced aberrations 45% 40% 35% 30% 25% 20% 15% 10% 5% 0%
1
2
3
4
5
ALL (n=241)
6
7
8
9
10
CLL CLL ALL (n=3046)
11
12
13
14
15
16
17
18
19
≥20
AML AML (n=2880)
MLL data
Number of unbalanced aberrations 45% 40% 35% 30% 25% 20% 15% 10% 5% 0%
1
2
3
4
5
ALL (n=241)
6
7
8
9
10
CLL CLL ALL (n=3046)
11
12
13
AML MDS AML (n=2880)
14
15
16
17
18
19
≥20
MDS (n=3487)
MLL data
Number of imbalances per tumor
Höglund M et al., GCC 2001
LOH on chromosome 5 and corresponding gene expression in a case with 5q deletion
Gene dosage effect on gene expression in AML with +8, +11, +13, -7, and del(5)(q13q31) sole vs. AML with normal karyotype Ratio 1,8 1,6 1,4 1,2 1 0,8 0,6 0,4 0,2 Chr. 8 AML +8 vs. AML-NK
Chr. 11 AML +11 vs. AML-NK
Chr. 13 AML +13 vs. AML-NK
Chr. 7 AML -7 vs. AML-NK
Chr. 5q13q31 AML del(5)(q13q31) vs. AML-NK Schoch C et al., Leukemia 2005
Effect of aneuploidy in combination with molecular mutations
Mutation of JAK2
Chromosomes 9
Effect of aneuploidy in combination with molecular mutations
X
Mutation of TP53
Chromosomes 17
Complete loss of TP53 function
Different types of chromosome aberrations
Albertson DG et al., Nat Genet 2003
Chromosome aberrations Aneuploidy/numerical aberrations Gains of whole chromosomes: Trisomy Tetrasomy Loss of whole chromosomes: Monosomy
Structural aberrations Balanced aberrations: translocation inversion insertion Unbalanced aberrations: deletion, duplication, ring chromosome, translocation
Balanced translocation
46,XX
46,XX,t(8;21)(q22;q22)
Balanced rearrangements
DNA mRNA
AAAA
AAAA
Protein
Overexpression of a normal protein
Expression of a chimeric protein
t(8;14)(q24;q32) ->CMYC
t(15;17)(q22;q12) -> PML-RARA
inv(3)(q21q26) -> EVI1
inv(16)(p13q22) -> CBFB-MYH11
Molecular aberrations
base exchanges: JAK2, FLT3-TKD
insertions: FLT3-ITD, NPM1
deletions: IKZF1
duplications: MLL-PTD
Molecular aberrations base exchanges: JAK2, NRAS, FLT3-TKD
Molecular aberrations insertions: FLT3-ITD, NPM1 SP
TM
JM
TK1
KI
TK2
RNA JM
TM
TK1 exon 11
exon 10
11 F
M -C +
-
+ +
-
12 R
11 R
-
-
DNA
exon 12
+
+ -
-
-
+
-
+ -
- M
-350 bp WT -cABL
Molecular aberrations deletions: IKZF1, RUNX1
duplications: MLL-PTD, RUNX1
CML Chromosome aberrations Balanced rearrangements:
unbalanced aberrations:
t(9;22)(q34;q11)
+8 +der(22)t(9;22) i(17)(q10)
t(3;21)(q26;q22) inv(3)(q21q26)
Molecular aberrations base exchanges: KRAS, NRAS
deletions: IKZF1
insertions: TET2
duplications: WT1
Progression to BC in CML on the cytogenetic level
46,XX
46,XX,t(9;22)(q34;q11) 47,XY,+8,t(9;22)(q34;q11)
Progression to BC in CML on the cytogenetic level
MLL data
Progression to BC in CML on the molecular level
Grossmann V et al., Leukemia 2011
Summary • Aneuploidy is a frequent phenomenon in leukemia • Different mechanisms lead to aneuploidy • Aneuploidy contributes to malignancy by –
deregulation of gene expression
–
loss of tumor suppressor genes
–
copy number increase of oncogenes
Munich Leukemia Laboratory
Cytomorphology Torsten Haferlach Johannes Schneider Eva Ammerlahn Sandra Eckstein Katharina Hermus Stefanie Jansa
Michele Kyncl Katja Macijewski Laura Urmetzer Jana Wagenbreth A.-Ch. Weichselbaumer
Immunophenotyping Wolfgang Kern Frauke Bellos Sabrina Baumann Angelika Eckelt Nicole Fietzeck Elena Gouberman Franziska Krakau
Rita Lapping Maja Lumpe Christiane Stangl Antonia Wagner Anja Winkler Carina Zanggl
Cytogenetics/FISH Claudia Haferlach Sandra Böker Silvia Finkböck Claudia Hepperger Jinda Holzwarth Doris Kestner Isabel Liebhart Verena Patterer Sarah Volkert Juliane Brendel Anna-Maria Brucks Sabina Bursch Jennifer Eland Annika Eßer Eyleen Haubner Annette Königsbauer Ulrike Schulz Kathrin Stadler
Marita Staller Susann Marschner Alida März Alexandra Mohr Nicole Mudrack Katja Niklaus Maximiliane Oppl Isabel Penzler Sandra Pöstgen Cornelia Pröger Karin Seige Franziska Schäfer Nadine Schedler Susanne Schmelz Carina Schreck Martin Schröter Melanie Zenger Kathleen Zieschang
Moleculargenetics Susanne Schnittger Frank Dicker Christiane Eder Annette Fasan Sabine Jeromin Simone Weber Katharina Bayer Jessica Brust Meike Büchler
x
Rabea Konietschke Julia Niederreiter Carina Schrauder Elisabeth Sirch Antje Stiel Madlen Ulke Nicole Wendland Christian Wittmann
NGS/Microarrays Alexander Kohlmann Vera Großmann Niroshan Nadarajah Sandra Weißmann Lucia Böck Katrin Butschalowski
Organisation Tamara Alpermann Peter Baumann Dirk Bursch Elke Erhard Claudia Eßbauer Julia Hägel Katrin Ilzhöfer Franziska Janou
Sandra Nawroth Franziska Prantl Tina Riedel Ulrike Riese Sandra Schulze Sabrina Weiß Wenke Worseg
Quality Management
Janine Elosge Stefan Harbich Franziska Pötzinger Sonja Schindela Elisa Stopp Melanie Wild
Monika Trnobransky
Number of balanced aberrations
Number of balanced aberrations 45 40 35 30 25 20 15 10 5 0 1
2 ALL
3
4
Number of balanced aberrations 45 40 35 30 25 20 15 10 5 0 1
2 ALL
3 CLL
4
Number of balanced aberrations 45 40 35 30 25 20 15 10 5 0 1
2 ALL
3 CLL
AML
4
Number of balanced aberrations 45 40 35 30 25 20 15 10 5 0
1
2 ALL
3 CLL
AML
MDS
4
Regions on chromosome 5 showing a significant lower gene expression in a case with 5q deletion
Unbalanced abnormalities Trisomy 8
5q deletion
Monosomy 7
der(1;7)(q10;p10) -> +1q,-7q
Effect of aneuploidy in combination with molecular mutations
Mutation of JAK2
Chromosomes 9