Cell trafficking in and out of
the bone marrow in MM
Irene Ghobrial, MD
Assistant Professor of Medicine
Harvard Medical School
Dana Farber Cancer Institute
Boston, MA
Model systems of studying MM
Trafficking and
Genetic and
dissemination of
it
epigene i
cells to new areas
tic
areas
regulation of
in the BM
MM cells
Growth
Survival
BM
Drug resistance
MM
Adhesion molecules
stromal cells
Cytokines
Interactions with OBL,
OCL, mesenchymal cells
Factors that
that regulate
regulate cell
trafficking
· Cell trafficking:
Homing
Adhesion
Egress or mobilization
· Variables that regulate cell trafficking
gg
MM cell intrinsic variables
Niche-intrinsic variables
SDF-1/CXCR4
· Chemokines play an important role in trafficking of
lymphocytes
· Receptors are CXCR4 and CXCR7
· Previous st d
u i
dies have demonst t
ra ed a
l
ro e f
o CXCR4
in
Prognosis
Migration
Proliferation
Adhesion
· AMD3100 induces egress of stem cells in clinical
trials
CXCR4 and CXCR7 dependent migration in MM
SDF-1
Mobilization of MM cells by AMD3100
MM.1S
CD138+
CD34+
MM.1S
CD34+
*
*
300
250
l)
250
ro
l)
ro
200
ont
ont
fC
200
C
o
f
%
o
(
150
%
150
lls
(
e
lls
C
e
C
100
100
c
ting
g
a
tin
ulc
la
rc
u
50
Ci
rc
50
Ci
0
0
02
6
12
34
Ti m e ( h r )
Time (Da
(D ys
y )
s
Control
150
AMD3100 50M
its
Bortezomib 10nM
125
Combination
Ungn
100
rmi
Fo
ny
75
loo
C
50
of
berm
25
Nu
0
EGM
M
GEMM
Total
Adhesion
· There are four major families of cell
adhesion molecules.
· CAMs, integrins, cadherins, and
selectins.
· Rolling is the first step of adhesion
mediated by selectins
Expression of selectins ligands on MM cell lines, patient samples and normal
subjects, detected by flow-cytometry using recombinant selectins.
U266
NCI
MM1S
Selectin receptors
were minimally
expressed on MM
cells
Pat. Sam.-1
Pat. Sam.-2
Pat. Sam.-3
Pat. Sam.-4
untsoC
Nor.
Nor Sub.-1
Nor.
Nor Sub.-2
Nor.
Nor Sub.-3
Control
E-Selectin Ligands
L-Selectin Ligands
P-Sl
Sel
i
ect n Li
Ligands
Fluorescence Intensity
The effect of a pan-selectin inhibitor on chemotaxis of MM cells in response to SDF1
Trans-Endothelial
)l 140
14
ro
Trans-well
120
120
)
ontc
rol
100
100
of
ontc
%
80
80
(
of
s
(%
lls
60
60
e
sll
c
e
40
tc
40
n
ting
re
a
e
20
20
hd
igr
A
M
0
0
0
100
10
250
25
500
1000
100
0
100
250
500
1000
Compound-A (uM)
Conc. uM
SDF 0
SDF 30 nM
nM
Coculture with HUVEC
-
+
+
+
+
+
Comp-A mcM
0
0
100 250
500 1000
Control
160
100 mg/Kg
140
p-P38
25 mg/Kg
lls 120
e
p-ERK
cg 100
Tubulin
tina 80
ul
irc
p-FAK
60
c
of
40
%
Tubulin
20
p-Src
0
Tubulin
5
1015
20
253035
40
4550
Time after cell Injection (min)
Genetic and epig
pgenetic
regulation intrinsic to MM
cells regulates homing,
gg,
adhesion and mobilization
miRNA 15-a and 16-1 regulate NFkB in tumor cells,
angiogenesis and tumor growth in vivo
B
no ) 400
tiav trol
ti
300
c
nt
a
oc 200
BkF ofd 100
N5 fol( 0
p6
e
-
rol
NF
t
bl
6-
6-
e
le
e
b
NF
5a1
R
NF
i
1
1
F
n
T
b
b
T
-1
-1
l+
m
m
iR
T
N
co
ra
a
e+
-m
+
iR
1
iR
T
tro
pro
-m
n
sc
+
scr
rob
pre
15a
e-m
e-m
1
co
p
pre
pr
pr
Regulate VEGF
secretion and
angiogenesis
In vivo tumor
growth
Hypoxia in the Bone Marrow regulates
MM1S
RPMI
H929
RPMI8226
H929
N
H
N
H
N
H
HIF-1
120 kD
-Actin
45 kD
HIF-2
118 kD
-Actin
45 kD
s
MM1S
OPM1
Count
CR4XC
Counts
HIF-1 alpha
LA4V
Conclusion
· MM cell trafficking:
Intrinsic factors within MM cells
Niche-specific factors
· Trafficking:
Homing, adhesion and mobilization
· Ud
Understandi
ding h
t ese
h
pat ways can l
d
ea to
novel therapeutic agents
Overcome resistance
Prevent dissemination
Prevent progression
Acknowledgement
· Dana-Farber Cancer Institute
Aldo Roccaro, MD, PhD, Kareem
Kareem Azab PhD
,
, Judith
Judith Runnels, PhD
PhD, Antonio
Sacco, Xavier Leleu, MD,PhD, Feda Azab, Hai Ngo, BS, Xiaoying Jia, BS, Nick
Burwick, MD, Phong Quang, Molly Melhem,
Ken Anderson, Nikhil Munshi, Barrett Rollins, Andrew Kung
· MGH: Charles Lin, PhD
R01CA125690, R01CA152607, Leukemia and
Lymphoma Society