The
Th SCID Hd
-Hu mod
mo e
d l f
o
multiple myeloma
Joshua Epstein
Contributors Past
Past and Present
Post docs:
Research staff:
MIRT Faculty:
James Chen
John fr
f eeman
r
III
Bart Barlogie
Ryuichi Fujii
Ling Liu
JC
James
J
h
JCChen
Hiroyuki Hata
Yu
Y xin Ze
Z ng
e
Igor Entin
M. Teresa Petrucci
Sean Fo
F ntenot
John Shaughnessy
Xavier Thomas
Aminah Henderson
Shmuel Yaccoby
Shmuel Yaccoby
Susan Mahaff
M
e
ahaff
Mahaff y
e
Shmuel Yaccoby
ey
Frank Zhan
Kenichiro Yata
Hannah Prat
Pr t
at
Lei Yin
Jonathan Stringer
Michele We
W zeman
e
MIRT clinical faculty and staff
HuiQing Xiao
Ronald Walker
NCI, MMRF, IMF, CRFA
MRI Focal Lesions:
Often Site of Relapse
Decrease in #
Hypo-intense
Early Relapse w/
and Size of
No Focal
Reappearing
Fl
Focal L
i
es ons
Li
Lesions
Fl
Focal L
i
es ons
`MRI-CR'
Sept 00
June 01
Dec 02
MRI Focal
Focal Lesions slow resolution
and site of relapse
Ev
· en
Ev
in complete
re
r mission, fo
f cal
o
lesions re
r tain
New
"dormant" my
m eloma
y
cells.
Recurrent
Baseline
CR (+240 days)
Relapse (+610 days)
MRI
MRI--Focal
Focal Lesions
Lesions--Cause
Cause of
Osteolytic Lesions
Expansion of
o
of
MRI
MRI
MRI--FL
FL with
disease
progression
T1--w
wt
STIR-wt
STIR
STIR--wt
wt
Corresp
Corres onding
pg
CT & PET
PE --CT
C
expansion of
osteolytic
lesion
4
4--28
28--05
05
8
8--25
25--05
05
MRI & CT
MRI & PET--CT
CT
My
M eloma
y
- Associated
A
Osteolytic
y
Lesions do not Heal
Lack of healing exhaustion of
the microenvironment?
Dormant Myeloma Cells?
CT take
tak n
CT take
tak n after 4 ye
y ars in
At
AAt diagnosis
complete re
r mission
"Empty" Osteolytic Lesion:
Lesion:
Exhaustion of Microenvironment?
Osteolytic lesion in newly
newly--diagnosed
diagnosed
patient: Devoid of myeloma cells?
CT
PET-CT
MRI
Dilemmas Prompting Development of
the SCID-
SCID hu Model for
for Primary Human
Human
Myeloma
Identity of the cells responsible for:
Myeloma growth
ggrowth
Relapse
Rl
Rol
o e f
o th
tthe
th microenvironment.
SCID
SCID--hu
hu Host System
y
for Primary
yy
Human Myeloma
hu
SCID
My
M eloma
y
Plasma Cells Proliferate
in the SCID
SCID--hu
hu Host
Plasma
Pl
Celllll-Dl
Deple
l t d
e
Purified Plasma Cells
Myeloma Bone Marrow Cells
Bone
Bon Marrow Cells
IgA Myeloma
My
IgA Myeloma
My
IgA Myeloma
My
800
800
800
Ig
Ig
Ig
Ig
Ig
Ig
600
IgG
600
IgG
600
IgG
IgA
IgA
IgA
(g/ml)
g/ml)(
(g/ml)
Ig
Ig 400
400
Ig 400
Human 200
Human 200
Human 200
0
0
0
0
02
24
46
68
810
10 12 14 16 18 20 22 24 26
0
02
24
46
68
810
10 12 14 16 18 20
0
02
24
46
68
810
10 12 14 16 18 20 22 24 26
Weeks From Inoculation
Weeks From Inoculation
Weeks From Inoculation
(Blood 92:2908, 1998: Blood 94:3576, 1999)
My
M eloma
y
Cell Growth Restricted
to the Human Microenvironment
Ct
Contro
t l
Ml
Myeloma
(Blood 92:2908, 1998: Blood 94:3576, 1999)
Sustained Proliferation
PProliferation of
of
Primary Myeloma Plasma Cells.
1
10
00
00
0
1st passage
ll))
2nd passage
/m
3rd passage
1
10
00
0
g(
inahacch 101
th
lliigghadabd 1
mbaamL
0
0..1
1
0
03
36
69
91
12
21
15
51
18
82
21
12
24
42
27
7
We
W e
e k
e s
k fr
f o
r m
o
in
i o
n c
o u
c l
u a
l t
a i
t o
i n
o of
o my
m e
y l
e o
l m
o a
m ce
c l
e l
l s
l
(Blood 92:2908, 1998: Blood 94:3576, 1999)
Summary
Myeloma plasma cells produce myeloma in
SCID
SCID--hu
hu mice.
Myeloma plasma cells are capable of
sustained proliferation.
MM PC do not change
cchange phenotype
pphenotype after
sustained growth.
No other cell
ccell in
in the
tthe bone marrow/blood of
patients can or is required for myeloma to
develop.
Conclusion
The my
m eloma
y
plasma cells contain the
prolif
pr
er
olif ativ
er
e
ativ cells
cells ("stem
(stem cells"?)
?)
? that
sustain the disease and determine its
course .
My
M eloma
y
p
ypro
r lifer
lif ativ
er
e
ativ cells: A defined
compartment (asymmetric cell
division ="stem cells")o
) r
or "re
r cycled
cy
"
my
m eloma
y
plasma cells?
The
T
debate persists...
B cells from patients give rise to clonal
myeloma
y
in NOD/
NOD
y/SCID mice (Pilarski).
CD19+ CD138
CD138-- myeloma
myeloma cells
expressing Hedgehog signaling
signaling, are
are the
clonogenic cells, and form myeloma in
NOD/SCID mice (Matsui
)
2007 .
)
BUT...
BUT
But...
CLL cells can be induced to differentiate into
monoclonal plasma cells (Tottermann 1980, cossman
1982, Srivastava 1984).
Embryonal stem cell
cell--associated
associated SOX2
expressed by CD138
CD138-- and
and CD138+ myeloma
cells (Dhodapkar 2007).
Myeloma
My
"stem cell": A defined entity
entit
or "re
r cycled
cy
" my
m eloma
y
myeloma plasma
plasma cells?
cells?
Tumor
T
microenvironment
Myeloma growth in the SCID
SCID--hu
hu model:
restricted to the human microenvironment.
Induces changes in the human bone ME:
Angiogenesis
Loss of osteoblasts
Osteoclastogenesis
Myeloma Changes the Bone
Bone Marrow
M
Marrow
Microenvironment
Osteoblast apoptosis
Osteoclastogenesis
At
Are h
these h
c anges
i
requ
d
re
Angiogenesis
Are these changes requi
for myeloma
m
myeloma cell survival?
ssurvival?
Endostatin Gene Therapy
in Myelomatous SCID
SCID--hu
hu host
AdEndo 2
5.3 g/ml
Control 2
AdEndo 1
1.7 g/ml
Control 1
M.W.
97 52 38 21
19
2
25
50
0
1
16
60
0
C
Co
on
nttrro
oll
A
Ad
dE
En
nd
do
os
stta
attiin
nIIn
njje
ec
cttiio
on
n
C
Co
on
nttrro
oll
A
Ad
dE
En
nd
do
os
stta
attiin
A
AdE
dn
nd
do
os
stta
attiin
n
2
20
00
0
12
1 0
)
2
l
)l
/mmgg/
/mml
1
15
50
0
(
gg/(
IIgg(
Igg(
8
80
0
d
A
AE
En
nd
do
os
stta
attiin
n
nan
nI
1
10
00
0
an
mma
ma
u
uum
H
H
4
40
0
5
50
0
0
0
0
01
12
23
34
45
56
67
78
89
91
10
01
11
11
12
21
13
3
0
01
12
23
34
45
56
67
78
89
9
W e e k s fr o m In tro d u c tio n o f M y elo m a C ells
W eek s fro m in tro d uc tio n of m y e lo m a cells
Osteoclasts and Myeloma
Pre Rx
Final
5
50
00
0
8
80
0
M
Me
ed
diia
an
nL
Le
ev
ve
ells
s
4
40
00
0
)
Untreated
2
l)
6
60
0
m
mml/
CP /m
g 300
(
c
n(
tii
Igg
4
40
0
otto tss
nI
pt
sst
Zoledronate
2
20
00
0
an
oop
al
ma
p
cl
um
AAp
ooc
HHu
2
20
0
e
100
%
t
%
s
O
0
0
H
Hu
um
ma
an
nIIg
gA
Ap
po
op
ptto
ottiic
cP
PC
CO
Os
stte
eo
oc
clla
as
stts
s
C
Co
on
nttrro
ollP
Pa
am
m..C
Co
on
nttrro
ollZ
Zo
oll..
P
Pa
am
miid
drro
on
na
atte
e
Z
Zo
olle
ed
drro
on
niic
cA
Ac
ciid
d
(Br.J.Haematol. 116:278, 2002)
Myeloma Requires Osteoclasts
Inhibition of osteoclast formation
has a profound anti myeloma effect
Start
7000
RANK-Fc
200
Control
60
6 0
0 0
0
160
) 5000
ml
120
4000
g/(g 3000
80
nIa 2000
m
RANK-Fc
40
Hu 1000
0
0
0
1
234
5
6
7
8
9
10
01
2
3
4
5
6
7
89
10 11
W ee k s
We
W e
e ks
(Br.J.Haematol. 116:278, 2002)
Osteoclasts and Myeloma:
Extramedullary disease
Pre Rx
Final
Extramedullary myeloma does not
require osteoclasts for survival and
proliferation
5000
Myeloma
Control
4000
Zoledronate
)l
m
g/ 3000
(
gI
n 2000
ma
Myeloma
uH
Myeloma
t
S a
t r
a t
+Zoled.
1000
Zoledronate
0
0
2
4
6
8
101214
1618
20
Weeks fr om introduction of myeloma cells
(Br.J.Haematol. 116:278, 2002)
My
M eloma
y
Cell Osteoclast
Co
Co--Culture
Culture
Ot
Osteoclas
l ts + Mye
M
loma
l
Cell
llls
ll
Osteoclast
precursors
pr
Myeloma Cells Alone
Osteoclasts
Osteoclast
Bone
Resorption
Resorption
Pits
(Cancer Res. 64:2016 2004)
Osteoblast restoration
rrestoration affects myeloma
myeloma
and bone remodeling
600
Human Ig;
Human BMD
x 500
-RR
400
prepre-
300
m
fro 200
100
angehc 0
%
--100
100
Control
Response
p
No Response
p
n=5
n=5
n=4
Haematologica.91:192,2006
PTH enhances
enhances myelomatous human
bone formation in SCID
SCID--hu
hu mice
Contro
Contr l
PTH
RANK--Fc
Fc
PTH + RANK--Fc
Fc
bone
matrix
Tet 1
Tet 2
The
T
SCID
SCID--hu
hu Model: Summary
Myeloma plasma cells (CD19- CD138+)
are capable
capable of sustained proliferation
Myeloma-associated changes in the
bi
bone marrow microenvironment are
essential for disease progression.
Other studies using the
SCID
SCID--hu
hu model
Variations using cell lines to study
homing to bone, drug
drug effects.
effects.