1. New and Emerging fields in Bio-medicine
Supporting Postgraduate Research with Internships in industry and Training Excellence
SPRITE : Workpackage 1
New and Emerging fields in Bio-medicine
Giovanna Muggiolu
Dietrich Walsh
Anne-Catherine Wéra

2. Workpackage 1: New and Emerging fields in Bio-medicine
This Work Package is led by CNRS and aims to push the frontiers of biomedical application of ion beams, in particular it targets both the treatment of cancer as well as detectors for accurate dosimetry control. By linking each of the individual training plans with a clinical or industrial environment SPRITE will ensure that the “real world” objectives of the research are translated into the clinical environment for the benefit of patients. This Work Package is particularly timely, as treating cancer with ion beams (hadron therapy) is a rapidly developing field in Europe at the present time and there is an urgent need for individuals who have the multidisciplinary training in this field to take the technology forward.

3. BIO MEDECINE ? ESR Dietrich Walsh ER Anne-Catherine Wéra ESR
UBW
Munich, Germany ER
Anne-Catherine Wéra
University of Surrey
Surrey, United Kingdom
BIO MEDECINE
ESR
Giovanna Muggiolu
CENBG/CNRS
Bordeaux, France
ESR
Xxxx xxxxx
CEA
Caen, France ?

4. The use of charged particles for cancer therapy
In 2011, the World Health Organisation recorded 57 millions death worldwide with 15% due to cancer.
Main cancer treatments are:
Surgery
Chemotherapy
Radiotherapy
50% of patient will undergo radiotherapy during the course of their treatment.
> 95% of the case radiotherapy is carried on with X-rays
>> Heavy charged particles
Need for new therapeutic strategies

5. The use of charged particles for cancer therapy
Traditional radiotherapy has side effects (i.e. chronic inflammation)
Proton therapy advantageous due to Bragg peak
Focussed proton beams may reduce side effects common in traditional radiotherapy (Work in progress)
Carbon Therapy already in use in Japan (Gunma Heavy Ion Centre)

6. Targeted irradiation and comparison with broad beam irradiation of cells
Workpackage 1: Micro Beam
Allows the analysis of cellular damage response to highly localised areas
Single particle – single cell irradiation
Study of the hypersensitive response of lung tumour cells after low-dose irradiation with low LET protons

7. Targeted irradiation and comparison with broad beam irradiation of cells
A549 cells irradiated with 4 MeV proton broad beam (10keV/µm)
as/ar=14 ± 6.7
77% survival (HRS): RBE=10.7 ± 3.3
( 10% survival: RBE= 1.9± 0.4)
>> 10 times lower doses for the same effect
End of HRS between 0.15 and 0.20 Gy

8. Targeted irradiation and comparison with broad beam irradiation of cells
Micro Beam Irradiation
A549 cells irradiated with 3.8 MeV proton micro beam (12keV/µm)
as/ar=47 ± 14
77% survival (HRS): RBE=41 ± 10
End of HRS at 10 hits

9. >> Poisson Statistics?
Targeted irradiation and comparison with broad beam irradiation of cells
? Threshold dose or Progressive transition ?
>> Poisson Statistics?

10. End of the HRS between 0.15 – 0.20 Gy But, due to Poisson statistics
Targeted irradiation and comparison with broad beam irradiation of cells
End of the HRS between 0.15 – 0.20 Gy But, due to Poisson statistics
0.15 Gy: some cells will receive a dose higher than 0.20 Gy > Outside the HRS
0.20 Gy: some cells will receive a dose smaller than 0.15 Gy > Inside the HRS

11. (micro beam parameters)
Targeted irradiation and comparison with broad beam irradiation of cells
Broad beam surviving fraction extrapolation from micro beam data and Poisson statistics
Poisson
distribution
Induced Repair Model
(micro beam parameters)
>> Poisson statistics can explain the variation in the threshold dose for HRS from broad beam to micro beam irradiation

12. Radio-sensitivity of sarcoma cell lines
Sarcomas represent a heterogeneous group of rare tumours accounting for approximately 1% of adult cancers and with more than 50 histological subtypes Surgery is the primary approach to treating most sarcoma even though radiation therapy has also been used for those patients with residual tumours following surgery.
Specific point mutations
Specific chromosome translocations
Genetic prospective
Many limited amplifications
Complex genomic profile

13. Centre d’Etudes Nucleaire Bordeaux-Gradignan
Interdisciplinary research team from Physics, Chemistry and Biology
IPCV
Supervisor: H. Seznec
Interdisciplinary Applications of Ions Beam in the Aquitaine Region
AIFIRA

14. Radio-sensitivity of sarcoma cell lines
Institute Bergonié
Treatment
Regional Cancer Centre
Teaching
Research
Sarcoma team
Unit INSERM U916 VINCO
Leader Research: F. Chibon
40 sarcoma cell lines from patients which are highly-well characterized from genetic/genomic and phenotypic point of view.

15. Radio-sensitivity of sarcoma cell lines
Sarcomas lines well-characterized
IB 115
amplifications of MDM2 and CDK4 genes
IB 106
unclassified sarcomas from paravertebral mass with heterogenic pleomorphic cells

16. Radio-sensitivity of sarcoma cell lines
GOAL
Evaluate the radiation sensitivity of sarcoma cell lines
Are sarcoma cells sensitive to photon therapy?
Are they sensitive also to proton therapy?
Have these two kind of radiations the same efficacy?
Comparing the radiation sensitivity of these cell lines using a photon facility (normally used for patients’ treatment) and a proton micro-beam (useful to study the proton-therapy)

17. Radio-sensitivity of sarcoma cell lines
EXPECTATIONS
SHORT TIME
Selection of Sarcoma cell lines according to their genetic point of view
Definition of an irradiation protocol from broad beam to proton beam (define irradiation mode, irradiation dose and radiation types)
Selection of biological end-points
Comparison Proton vs X-rays irradiation
LONG TIME
Development and validation of in vitro irradiation protocols combining both protons and nanoparticles

18. Role of mitochondria in the cellular response to ionising radiation
What are Mitochondria?
Mitochondria are biological „powerplants“. There are per cell and they range from 500nm-2µm
Main source of energy in a cell (Energy = ATP)
More energy a cell requires, more mitochondria it has
Cells can not survive without mitochondria!
Mitochondria have a double membrane which they use to keep a constant ΔV between mitochondria and inside of the cell (polarisation). This is required for function
Mitochondria have their own DNA which is inherited from the mother.

19. Role of mitochondria in the cellular response to ionising radiation
Roles of the Mitochondria in cells
Mitochondria are the energy factories of cells
Use sugar to produce cellular energy (ATP) the process requires oxygen and mitochondria (Slow rate, high ATP yield)
Mitochondria regulate energy „metabolism“ of cells
Cancer cells require high amounts of energy
Mitochondrial disfunction is involved in many diseases

20. Role of mitochondria in the cellular response to ionising radiation
To identify biological effects of microbeam irradiation on Mitochondria.
Initial Ideas:
Microbeam irradiation of Mitochondria in live cells (novel techniques employed)
Live cell imaging of mitochondrial depolarisation (Does pinpoint radiation destroy mitochondria?)
Analysis of gene expression level of mitochondrial proteins (qRT-PCR)
Imaging of DNA strand breaks in mitochondrial DNA

21. Role of mitochondria in the cellular response to ionising radiation
SNAKE
Superconducting nanoprobe
for applied nuclear physics experiments
Up to 25 MeV protons
Lateral resolution of 1µm

22. Summary
Diverse projects in the field of radiation biology and biomedicine
The goal is to bring together physicists and biologist in a clinical and industrial environment
The research being conducted aims to answer emerging questions in the field of experimental radiation oncology which are relevant for future particle therapy.

23. THANK YOU FOR YOUR ATTENTION!!!
SPRITE : Workpackage 1
New and Emerging fields in Bio-medicine
Giovanna Muggiolu
Dietrich Walsh
Anne-Catherine Wéra