molecular biomarkers in radiotherapy of cervical cancer a collaboration project between department...
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Molecular Biomarkers in Radiotherapy of Cervical Cancer
A collaboration project between
Department of Gynecologic Oncology
and
Department of Radiation Biology
Project group
The Radiation Therapy Team at Dept. of Gynecologic Oncology /Gunnar B. Kristensen, Dr. MedThe Clinical Radiation Biology Group at Dept. of Radiation Biology /Heidi Lyng, Dr. Philos
Radiation field
Lymph nodes
External irradiation: Tumour region (50 Gy) and the rest of pelvis (45 Gy) Endocavitary brachytherapy: Cervix (21 Gy)
Narrow therapeutic window high frequency of side effects to pelvic organs
Radiotherapy of cervical carcinomas
External radiation, field_1 front
Need for improved treatment – more individualized therapy based on biological information
Aims
Molecular methods based on microarrays will be combined with MR and eventually PET techniques to find biomarkers that can be
utilized for biologically optimized therapy
Identify predictive biomarkers for the therapeutic outcome, including patient survival, locoregional tumor control and normal tissue side effects.
Identify key radiation regulated pathways in tumors and possible targets for molecular intervention.
Explore how the molecular findings can be combined with functional (MR and PET) and molecular imaging in treatment planning and response monitoring.
Microarrays MR imaging MR-spectroscopy
Study protocol on cervical cancer, stage 2b-4a
Tumor biopsies
DCE-MRI
Blood sample
DCE-MRI
PathologyMR/CT findingsRadiation fieldFollow-up
MedInsight
Clinical data base
Tumor biopsies
Radiation therapy, curative intent
Research projects
T2-MRI
CT dose plan
Image storage Blood and tissue storage
>300 patients included
Research projects
Studies in cell linesMolecular screening Signaling
Tumor biopsies
Functional imaging
DCE-MRI
Normal tissue side effects
CT dose plan, blood samples
Molecular screening - gene profile associated with clinical outcome - basis for further molecular studies
Collaboration with statisticiansFrigessi, Glad, Holden: UiO, NRVan de Wiel, Vrije Universiteit, Amsterdam
Frigessi et al, Nucleic Acids Res, 2005Scheel et al, Bioinformatics, 2005Ferkingstad et al., Genome Biology, 2008Nygaard et al., BMC Genomics, 2008
”New” genes
Marker for clinical outcome?Terapeutic target?
P = 0.02
Gene profile 1
Gene profile 2
Classification of patients with different outcome based on gene profile
Genuttrykk (mRNA)
Analysis of tumor biopsies
PhD student Malin Lando
-1.5 -1.0 -0.5 0.0 0.5 1.0 1.5
0
1
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-1.5
-1.0
-0.5
0.0
0.5
1.0
1.5F
requ
ency
(%
)
aCGH ratio (log2)
aCG
H r
atio
(lo
g 2)
Chromosomal location 1pter-Xqter
AC024/01DNA index: 1.00Tumor cell fraction: 55%
B
1 3 5 7 9 11 13 15 17 19 21 X 2 4 6 8 10 12 14 16 18 20 22
1&2
1
2
3
2&3
1&2
2&3
1&2
Molecular screening: intratumor heterogeneity in gene copy number
Pronounced heterogeneity in copy number within cervix tumors → resistent subpopulations may emerge at later stages of the disease
Genome wide screening of copy number in cervix tumor
-1pcen-31+1q-6q24-25-12q-17p-19p
22% 34% 44%
Evolution
-1pcen-31+1q-6q24-25-12q-17p-19p
-13qcen-34 -13qcen-34
-1p31-ter-2q21-ter-4p+6p-8p
C
-1pcen-31+1q-6q24-25-12q-17p-19p
Lyng et al., Genome Biology 2008
Subpopulations oftumor cells with differentgenetic characteristics
Characterization of signaling pathways of importance for outcome → mechanisms of activation
Screening signaling
MSN
P = 0.004
Pro
gre
ssio
n f
ree
su
rviv
al
Score N Events
0-6 87 379 72 15
Time (months)
Lyng et al., BMC Genomics, 2006
Low MSN expression → poor outcome
Low MSN expression
High MSN expression
Analysis of tumor biopsies
PhD student Cathinka Halle
Collaboration with Division of PathologyRuth Holm
Metabolic screening by use of MR-spectroscopy - relationship to molecular data, imaging (MR) and clinical data
Collaboration with the MR center in TrondheimIngrid Gribbestad
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CreGPCPC
ChoCre
-Glc
Lac
Lac
TSP
FA
– C
H3
FA
– (
CH
2) n
FA
–C
H2
– C
H3
FA
– C
H2
– C
H2
– C
O-
Ac
Gly
cero
l bac
kbon
e
Tau
FA
– C
H =
CH
– C
H2
– C
H2
Gly
Tumor biopsy from cervix cancer Metabolic profile
Lyng et al., BMC Cancer 2007
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0
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-20 0 20 40 60 80 100120 140160 180200
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Measured apoptotic cell density (cells/mm2)
r = 0.95p < 0.001
Pre
dict
ed a
popt
otic
cel
l den
sity
(ce
lls/m
m2 )
B
Metabolites involved in treatment induced apoptosis