Radiation-associated and sporadic UPS/MFH : Searching for novel therapeutic strategies
description
Transcript of Radiation-associated and sporadic UPS/MFH : Searching for novel therapeutic strategies
Sarcoma Research LaboratoryUniversity of Texas, MD Anderson Cancer Center
Radiation-associated and sporadic UPS/MFH: Searching
for novel therapeutic strategies
Keila Enitt Torres
UPS/MFH Pleomorphic malignant fibrous
histiocytoma (MFH) unclassified pleomorphic sarcoma
Current technology ? definable line of differentiation
UPS Peak incidence in the 6th and 7th decades of
life Typically large, deep, aggressive tumors 5% - metastases at presentation (lung) <3% radiation-associated
UPS Treatment
Radical excision with negative margins Neo-Adjuvant RT is offered widely negative
margins is not possible. Chemotherapy Unresectable
disease/metastatic disease
5-year disease-specific survival 44- 66%*
Approach Low throughput: PI3K/AKT signaling pathway
Determine if AKT/mTOR axis is a relevant target for UPS therapy
High throughput: genomic/proteomic analysis Understand genetic and molecular de-regulations UPS progression
PI3K/AKT pathway
Nature Medicine 13, 748 - 753 (2007)
HGF p-c-Met p-MEK p-AKTc-Met
Goal #1 Confirm whether the AKT/mTOR axis is a potential
target for UPS/MFH therapy Determine whether this pathway is differentially
activated in radiation-associated versus sporadic UPS
Radiation-associated sarcoma Diagnosis of RAS based on the criteria
published by Cahan, et al. in 1948.* Previous radiation tx for different tumor Development of the sarcoma - radiation field Histologically different from the primary cancer Minimum latent period of 5 years between the
radiation and development of the sarcoma Arlen et al. 1971- modified definition
Latency period of 3 years
*Cahan et al. Cancer 1948;1:3-29.
Bioresources in the laboratory Cell lines Animal models Human Tissue
Frozen FFPE
Cell strain/cell line bank
> 60 human tumors
13 Sporadic UPS cell strains
6 Radiation-associated
UPS cell strains
RIS-819.1 RIS-023
UPS-060UPS-485 UPS-186
UPS-060
AKT/mTOR pathway is activated in UPS
UPS-186
RIS-D
L620
RIS-D
L618
hMSC
p4EBP1
pAKT
4EBP1
AKT
S6K
b-actin
UPS-060
UPS-356
UPS-DL5
11
RIS-02
3
RIS-81
9
Radiation-associated Sporadic
pS6K
Rapamycin
0 10 100
0 10 100
0 10 100
RIS -819RIS -023 UPS-485UPS-060
Radiation-associated Sporadic
0 10 100
p4EBP1
pAKT
4EBP1
AKT
S6K
b-actin
pS6K
Rapamycin
Control 0.05uM 0.1 uM 0.5 uM 1 uM 5 uM0.0
20.040.060.080.0
100.0120.0140.0
RIS-819
% C
ell P
rolif
erati
on
Con 0. 0. 0.1 u
M5 u
M0.0
20.040.060.080.0
100.0120.0140.0
RIS-023
%Ce
ll Pr
olife
ratio
n
Concentration (mM)
0 0.05 0.1 0.5 1 5
0 0.05 0.1 0.5 1 5
Radi
ation
-ass
ocia
ted
Concentration (mM)
0 0.05 0.1 0.5 1 50.0
50.0
100.0
150.0UPS-060
% C
ell P
rolif
erati
on
0 0.05 0.1 0.5 1 50.0
50.0
100.0
150.0UPS-485
% C
ell P
rolif
erati
on
Spor
adic
Concentration (mM)Concentration (mM)
pAKT
AKT
pS6KS6Kb-actin
p4EBP14EBP1
mM/4hr 0 0.05 0.1
0 0.05 0.1
UPS-060 RIS-819
PI103
PI103
0 0.05 0.1 0.5 10
20
40
60
80
100
120
RIS-819
% C
ell p
rolif
erati
on
0 0.05 0.1 0.5 10
20
40
60
80
100
120
UPS-485
% C
ell p
rolif
erati
on
0 0.05 0.1 0.5 10
20
40
60
80
100
120
UPS-060
% C
ell p
rolif
erati
on
Concentration (mM)
Radi
ation
-ass
ocia
ted
Concentration (mM)
Spor
adic
Concentration (mM)Concentration (mM)
0 0.05 0.1 0.5 1 50
20
40
60
80
100
120
RIS-023%
Cel
l pro
lifer
ation
Control PI1030
1020304050607080
G1SG2/M
Control PI1030
102030405060708090
G1SG2/M
RIS-819
Control PI103
UPS-060
Control PI103
Cell Cycle
Migration
Control PI103
RIS-
819
Control PI1030
20
40
60
80
100
120
% M
igra
tion
UPS
-060
Control PI1030
20
40
60
80
100
120
% M
igra
tion
Invasion
Control PI103
UPS-
060
Control PI1030
20
40
60
80
100
120
% In
vasio
n
RIS-
819
Control PI1030
20
40
60
80
100
120
% In
vasio
n
In vivo studies Orthotopic Xenograft Model
1-2mm pieces of tumor
0 4 8 12 16 200
100200300400500600700800900
1000ControlPI 50 mg/mL
Days
Tum
or V
olum
e (m
m3)
*
Control PI1030
0.20.40.60.8
11.21.41.61.8
2
Tum
or w
eigh
t (g)
*
RIS-819 tumor growth
pAKT p4EBP1 pS6RP Ki67H&E
PI10
3Co
ntro
lRIS-819 Immunohistochemistry
1 4 8 12 16 24 32 40 480
50
100
150
200
250
300
350
Control
PI103 (50 mg/Kg)
Days
Tum
or V
olum
e (m
m3)
UPS-060.1 tumor growth
PI103Control
*
pAKT p4EBP1 pS6RP Ki67H&E
PI10
3Co
ntro
lUPS-060.1 Immunohistochemistry
Summary PI3K/AKT pathway is activated in radiation-
associated and sporadic UPS RA-UPS and sporadic UPS are sensitive to
PI103 Effects in vivo model => Cell death
Current studies
ProteomicsmiRNA/
non-coding RNAs
mRNAMethylation
DNA copy number variation
DNA Sequencing
Sequenome
Exon Sequencing
Array-comparative
genomic hybridization
miRNA analysis
RPPA
TMA
Reverse-phase protein lysate array for cell signaling analysis
EIF4E
29 molecules differentially
expressed
HER-2 SignalingERK/MAPK
PI3K/AKT/mTORErbB AMPK
HGF signalingIL-15 Signaling -3.5058 5.1031
(Log 2)
Mesenchymal cells
UPS
>50%P value <0.01
Unsupervised hierarchical clustering
Unsupervised hierarchical clusteringRA-UPS versus sporadic UPS
-3.5058 5.1031
(Log 2)
RA-UPS
Sporadic-UPS
Sarcoma Research LaboratoryUniversity of Texas, MD Anderson Cancer Center
Dina Lev Raphael E. Pollock Billy Wang Alexander J. Lazar Vinod Ravi
SARC (Sarcoma Alliance for Research through Collaboration)
Acknowledgments