Pathogenesis of Multiple Myeloma:
Ig translocations
CYCLIN D dysregulation
other oncogenic events
Michael Kuehl
MM is a post-germinal center PC tumor
Short-lived PC
3 days
IgM
Germinal Center
G,A,E,D
Lymph
Node
HYPERMUTATION
Lymphoblast
Y
Y
ANTIGEN SELECTION
IGH SWITCHING
YY
Mature B cell
VDJ recombination
Y Y
G,A,E,D
Immature
B cell
Plasmablast > Long-lived PC
> 30 days
Bone Marrow
Diversity of IgH translocations in MM
5 recurrent partners
40%
11q13 (CYCLIN D1)
15
6p21 (CYCLIN D3)
3
16q23 (c-MAF > CYCLIN D2)
5
20q11 (MAF B > CYCLIN D2)
2
4p16 (MMSET&FGFR3)
15
8q24 (c-MYC)
3%
Rare partners
17%
NO IgH translocation
40%
*MYC translocations: a late event in MM
Burkitt's
Myeloma
Timing
Early (initiation)
Late (progression)
Prevalence
Always
MGUS: absent or rare
All MM: 15% (IgH, 3%)
Advanced MM: 44%
HMCL: 91%
Type of translocation
Simple
Mostly complex
Ig locus associated
Always
~ 50% (IgH ~ Ig >> Ig)
B cell mechanism
Yes
No (off in PC & tumors)
Heterogeneity
No
Sometimes
* c-MYC >> N-MYC > L-MYC
Two kinds of chromosome content in MM
Hyperdiploid (HRD) & non-hyperdiploid (NHRD)
each ~50% tumors
NHRD poorer prognosis than HRD
HRD (48-75 chromosomes)
multiple trisomies involving 8 chromosomes
3, 5, 7, 9, 11, 15, 19, 21
NHRD (<48 and/or >75 chromosomes)
increased prevalence of IgH translocations
Translocations: HRD vs NHRD MM
(metaphase FISH analysis of 48 MM tumors)
HRD NHRD
IgH TLC
35
81
5 recurrent partners
12
63
other IgH partners
23
18
Ig TLC
15
18
MYC TLC
41
46
> 2 Ig translocations in a MM tumor cell
(58% of HMCL, 27% of advanced MM, ~5%MGUS)
Rarely involves 2 of the 5 major recurrent partners, but
this includes all functionally distinct combinations:
CYCLIN D1 or CYCLIN D3 + c-MAF or MAF B
CYCLIN D1 or CYCLIN D3 + FGFR3/MMSET
FGFR3/MMSET + c-MAF or MAF B
More often a recurrent partner and a rare partner, or
two different rare partners
Most often c-MYC and a recurrent or a rare partner
Different roles for translocations in MM
increased DNA labelling index
bone destruction
Hyperdiploid pathway
angiogenesis
(12% recurrent/1o IgH TLC)
Germinal
Intra-
Extra-
Myeloma
Medullary
center
MGUS
medullary
Cell Line
MM (PCL)
MM
(HMCL)
B cell
Non-hyperdiploid pathway
(69% recurrent/1o IgH TLC)
2o (Ig) TLC (IgH ~ Ig >> Ig)
MYC TLC
Cyclin D1, D2, D3 Relative RNA Expression in PB, PC, 261 MM, HMCL
8 MM tumor groups
PB PC
4p
MAF 6p
11q
D1
D1+D2
D2
O
HMCL
16%
7%
3%
16%
31%
7%
17%
2%
CCND1
CCND2
CCND3
8 TC (translocation/Cyclin D) groups
GROUP % MM %HRD
%MGUS
4p
16
29
0
MAF
7
16
9
6p
3
29
0
11q
16
3
29
D1
31
93
24
D1+D2
7
95
9
D2
17
40
29
none
2
33
0
TOTAL
261
50%
21
Limited # of samples
Specific gene expression patterns in 5 largest MM groups:
(maf > 4 > D1 > 11 > D2)
Supervised cluster
using 1002 genes
that best distinguish
the 5 largest MM
groups
Prevalence of K- and N-RAS mutations in 8 MM Groups
GROUP # MM % K-RAS P value % N-RAS P value % K- or N-RAS
4p
57
14
0.55
3
<.01
17
MAF
10
20
0.69
10
1
30
6p
1
0
-
0
-
0
11q
58
17
1
24
<.02
41
D1
73
16
0.71
21
0.04
37
D1+D2
10
40
0.07
0
0.36
40
D2
35
20
0.59
3
0.06
23
none
4
0
1
25
0.36
25
TOTAL
248
17
-
14
-
31
Selective progression of MM to HMCL
TUMOR
HMCL
5 recurrent IgH translocations
40%
76%
D1 group
35%
NONE
D1 + D2 group
8%
3%
A Translocation/ CYCLIN D (TC) classification of MM
(based on 5 recurrent Translocations & Cyclin D expression)
Analogous to classification of ALL based on translocations and ploidy
Based on apparent unifying and early dysregulation of a CYCLIN D gene
in MGUS and MM
Gene expression patterns associated with group
Biological and clinical features associated with groups
Prognosis and response to different treatments associated with groups
Tumor phenotype appears to be determined by early pathogenic events,
consistent with hypothesis that MM represents several different diseases
The Rb pathway can be altered by sequential
events in a MM tumor
maf
4p16
D2
D1
D1
11q13
6p21
16q23 c-maf
FGFR3
+
Hyperdiploid
CCND1
CCND3
20q11 mafB
MMSET
D2
p16
INK4a
p15
INK4b
NONE
Cyclin D2
Cyclin D1
Cyclin D3
p18 INK4c
CDK 4, 6
CDK 4, 6
p19
INK4d
CDK 4, 6
G1
S
Phase
phase
P
P
P
P
Rb
E2F
Rb
E2F
OFF
ON
increased DNA labeling index
bone destruction
angiogenesis
?
Germinal
Intra-
Extra-
Center
Myeloma
MGUS
Smoldering
medullary
medullary
B cell
Cell Line
Myeloma
Myeloma
Myeloma
Karyotypic abnormalities
1o Ig TLC
Secondary (Ig) TLC
del13
activating mutations: N, K- RAS, FGFR3
p18 deletion
Hyperdiploidy
MYC dysregulation
p53 mutation
COLLABORATORS
Kuehl lab (current)
Cornell Medical
Leslie Brents
Leif Bergsagel
Conny Cultraro
Marta Chesi
Ana Gabrea
U. Arkansas
Ben Ristau
John Shaughnessy
Adriana Zingone
Bart Barlogie
Jeff Sawyer
Kuehl lab (past)
Mayo Clinic
Amel Dib
Rafael Fonseca
Christopher Bean
U. Copenhagen
Tim Peterson
Thomas Rasmussen
Marina Martelli
NCI
Ying Qi
Lou Staudt
Yaping Shou
Lanny Kirsch
Laura Raducha-Grace
Thomas Ried
Siegfried Janz