Molecular Biomarkers in Radiotherapy of Cervical Cancer A collaboration project between Department of Gynecologic Oncology and Department of Radiation.

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

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1

2

3

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5

6

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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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cero

l bac

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Tau

FA

– C

H =

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H2

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Tumor biopsy from cervix cancer Metabolic profile

Lyng et al., BMC Cancer 2007

-50

0

50

100

150

200

-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