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The relative contribution of microenvironmental interactions to the regulation of gene expression programs in multiple myeloma cells
Liat Nadav
Molecular Cell Biology Department Weizmann Institute of Science
The Hematology InstituteTel-Aviv Sourasky Medical Center
Bentzi Katz, PhD
The Hematology InstituteTel-Aviv Sourasky Medical Center
Diversity – a hallmark of tumor progression Diversity – a hallmark of tumor progression
BM growth
Dissemination
Osteoclast activation
Drug resistance I
Drug resistance II
Drug resistance III
Diversity - is it a stochastic process, Diversity - is it a stochastic process,
or is there any direction, regulation ? or is there any direction, regulation ?
monocyte
lymphocyte
neutrophil
plateletsRBC
Bonemarrow
Bonemarrow
Regulation of Megakarypoiesis by BM nichesRegulation of Megakarypoiesis by BM niches (Rafii S. et al., 1994-2007)(Rafii S. et al., 1994-2007)
Soluble factorsCellular interactionsAdhesive interactions
Can adhesive interactions generate diversity of Can adhesive interactions generate diversity of malignant plasma cells ?malignant plasma cells ?
What is the physiological significance of such a process ?What is the physiological significance of such a process ?
Isolation of adhesive variantsIsolation of adhesive variantsfrom the ARH-77 linefrom the ARH-77 line
Type-F cells(floating)
Type-A cells(adherent)
No
. of
even
ts
101 102 103 104
CD138
Type-AType-F
Flow cytometry pattern
Type-A cells
Type-F cells
Differential gene expression in adhesive variantsDifferential gene expression in adhesive variants
I
II
III
IV
V
I
II
III
IV
V
5 individual enrichments RNA purification Affymetrix array
0
10
20
30
40
50
Per
cent
of
gene
s in
eac
h fu
nctio
n ca
tego
ry
Metabolism
Signal transduction
Transport
Differentiation andDevelopment
Transcription factors
Apoptosis
Cell cycle
Adhesion
Motility
Immune response
Undefined
Color Code
Type-A cells Type-F cells
Assignment of gene expressed in adhesive variantsAssignment of gene expressed in adhesive variants
Coding region
Regulatory region
Regulatory elements
Gene AGene BGene CGene D
RNA from cell I
Pathway X is active in cell I
Gene AGene BGene CGene D
RNA from cell II
Pathway Y is active in cell II
Signaling pathways
Promotor analysis
Promotor analysis
Type-A cells Type-F cells
?NFB (CCND1, CXCL11, IL6)
EGR(c-jun, EGR1, EGR2)
NFB1
SRF
NFB2
L1
L2L3
L4
L5L6L7
L8
L9L10
H1
H2H3
H4
H5H6
H7
H8
H9
0
2
4
6
8
10
12
14
0 2 4 6 8 10 12
Ave
rage
exp
ress
ion
in T
ype
F
Average expression in Type A
B-Cell differentiation state
HighLow
H1- GADD45AL1- TIMP1
H2- IgH VDJL2- BCL6
H3- IgH variableL3- IL-6
H4- Ig rearrangedL4- EGR4
H5- IgH HML5- TNF
H6- IgH JL6- TNFRSF2
H7- IgH G1-4L7- TNFRSF4
H8- IgH ML8- TNFRSF9
H9- IgH M1-2L9- MYCN
L10- BTK
Cell adhesion regulates differentiation of plasma cellsCell adhesion regulates differentiation of plasma cells
GAPDH
F A
IgJ
GAPDH
F A
EGR1
Type-A cells
AF cells
GAPDH
EGR1
IgJ
Type-A cells
AF cells
Regulation of gene expression is reversibleRegulation of gene expression is reversible
Lapsed time )days(
Tum
or F
ree
mic
e )%
(
0
20
40
60
80
100
20 40 60 80 100 120
Type-A cells
Type-F cells
Malignancy potential of plasma cell variants Malignancy potential of plasma cell variants
1. Microenvironmental interactions can generate diversity of malignant plasma cells
2. Directional diversity – differentiation
3. Specific putative targets: NFB, EGR )Type-A(
4. Type-A cells – are they represent “stem cells” or “stem state” ?
Phenotype control by microenvironmental interactions Phenotype control by microenvironmental interactions
CD138
CD
38
Co
ntr
ol
103
102
101
101 102 103
Mec
han
ical
+en
zym
es
103
102
101
Spicules
Patient BMs also contain diverse MM cellsPatient BMs also contain diverse MM cells
Fluid
MM bone marrow
Thanks…Thanks…
Benny GeigerMolecular Cell Biology Department
Weizmann Institute of Science
Tal Shay, Eytan Domany
Physics of Complex Systems Department Weizmann Institute of Science
Ella NaparstekShoshie Baron
The Hematology InstituteTel-Aviv Sourasky Medical Center