Promoting Osteoblastogenesis with a Novel clinical
grade Dkk-1 Neutralizing Antibody in the Treatment
of Multiple Myeloma Related Bone Disease
Samantha Pozzi, MD
Multiple myeloma is characterized by osteolytic bone lesions in
over 80% oft
f h
the pati
tients
This is related to an imbalance between osteoblasts and osteoclasts
PHYSIOLOGIC BONE
REMODELING
MCSF
DKK1
MIP-1
OPG
RANKL
osteoclast
osteoblast
MCSF
MIP-1
-
OPG
DKK1
RANKL
osteoclast
ostb
teo l
blast
N.Raje-S.Vallet
Current treatment of multiple myeloma bone disease is
mainly focused on osteoclasts
IL6
· BP
VEGF
G
BAFF
IGF1
RANKL · anti-RANKL
LFA-1
VEGF
VLA-4
IL6
VCAM-1
DKK1
MIP-1
ICAM-1
Bone marrow
stromal cells
Osteoblasts
Osteoclasts
No demonstration of bone healing
Modified from N.Raje-S.Vallet
Rational of the study
Dkk-1 is wnt-signaling antagonist, that inhibits osteoblasts differentiation
High
levels
levels of
of Dkk
Dkk-1 are
are expressed in
in a subgroup
subgroup of multiple myeloma
patients
High levels of Dkk1 correlate with bone lytic lesions in multiple myeloma
patients
NEJM 2003;349:2483-94
Anti-Dkk1 monoclonal antibody target osteoblasts enhancing osteoblasts
function
Targeting
Targeting Dkk1
Dkk1 in multiple myeloma
Myeloma Cells
MIP-1,
BAFF,
IL6,
MIP-1, IL6,
VEGF
VEGF RANKL
,
DKK1
X
OPG
m-CSF
fms
TRAIL-R
WNT
MAP
RANKL
K
ERK
TRAF6
JNK
p38
p50/p52
B-
catenin
AP-1
RUNX2
INTEGRIN
Osteoblasts
Osteoclast
Modified from N.Raje-S.Vallet
Preliminary results of an anti-Dkk1
monoclonal antibody
Effects of anti-Dkk1 on Osteoblasts differentiation
Alkaline Phosphatase Staining on day 28 in OB medium culture (4)
Alizarin red on day 21 in OB
OB medi
dium cult
lture
control
Anti-Dkk1
IgG 1000 ng/ml
10 ng/ml
100 ng/ml
1000 ng/ml
Alizarin Red & Alkaline phosphatase quantitative analysis in
osteoblasts differentiation and function
ALP in OB differentiation
Alizarin Red in OB differentiation
4
3
3,5
2,5
3
2
2,5
IgG
IgG
DKK10
gG
DKK10
2
/I 1,5
OD/IgG
DKK100
OD
DKK100
1,5
DKK1000
DKK1000
1
1
0,5
0,5
0
0
D3
D14
D21
D24
D27
D10
D17
D21
D24
D27
days
days
Effects of anti-Dkk1 on osteoclasts differentiation
and function
TRACP staining at day 28
28 in
in osteoclasts
osteoclasts culture
culture (4):
Pit formation assay at day 28 in osteoclasts culture
IgG 1000 ng/ml
anti DKK1 1000 ng/ml
Conclusions
Preliminary results in vitro show that:
anti-Dkk1 monoclonal Ab enhances osteoblasts diff
dif erentiation
f
and function, in a dose dependent manner
anti-Dkk1 monoclonal Ab inhibits osteoclasts differentiation and
function, in osteoclasts derived from the mononuclear fraction of bone
marrow aspirates of patients with multiple myeloma
Future directions of multiple myeloma treatment:
targeting the bone microenvironment
BAFF
anti-myeloma
treatments
VEGF
Myeloma cells
BAFFanti-Dkk1
· BP
BAFF
IL6
VEGF
·
VEGF
BAFF
anti-RANKL
IGF1
RANKL
VEGF
LFA-1
VEGF
VLA-4
IL6
4
VCAM-1
DKK1
MIP-1
ICAM-1
Bone marrow
stl
tromal celllls
Osteoblasts
Osteoclasts
Modified from N.Raje-S.Vallet