Public Security S&T Symposium 2009

1. Public Security S&T Symposium 2009 Rapid Identification of Radiologically-Exposed Individuals for Medical Casualty Management CRTI-06-146RD Presented by: Ruth Wilkins, Health Canada

2. Health Santé Canada Canada CANADIAN PARTNERS: Consumer and Clinical Radiation Protection Bureau, Health Canada Defence Research and Development Canada- Ottawa Atomic Energy of Canada Limited – Chalk River McMaster Institute of Applied Radiation Sciences, McMaster University

3. US PARTNERS: Armed Forces Radiobiology Research Institute, Bethesda, MD (AFRRI) Oak Ridge Institute for Science and Education, Oak Ridge, TN Litron Laboratories, Rochester, NY University of Rochester Medical Centre, Rochester, NY

4. PROJECT BACKGROUND: Goal: • To expand the National Biological Dosimetry Response Plan (NBDRP) from a proof-of-concept initiative into a formalized “Medical and Casualty Management Tool” Scope: • To enhance our emergency triage biodosimetry capacity by developing stronger linkages with clinical cytogenetic laboratories across the country and with our U.S. counterparts.

5. PROJECT BACKGROUND: Objectives: • conduct ongoing training and exercising of the four core laboratories • expand NBDRP linkages with our U.S. counterparts to work towards an integrated North American emergency response network. • develop and implement a training program into the curriculum for Medical Cytogeneticists at the Michener Institute (Toronto) • investigate and establish sustainable networks with the cytogenetic laboratories to encourage their participation in the NBDRP network • Continue research into novel methods for biological dosimetry and biomarkers of exposure

6. Why do we need biological dosimetry?

7. Normal DICENTRIC CHROMOSOME ASSAY (DCA): • Currently the gold standard • ISO committee published DCA standard in 2004 and mass casualty standard in 2008 • Look for unstable chromosome aberrations in white blood cells from an individual’s blood sample • Dicentric chromosomes • Ring chromosomes • Very sensitive and specific to ionizing radiation Ring Dicentric

8. LIMITATIONS OF DICENTRIC CHROMOSOME ASSAY • Dicentric chromosome assay is time consuming • For low dose assessments, (down to 0.15 Gy), 1000 metaphase spreads must be examined • 2 person days of scoring • In the case of a large scale exposure to ionizing radiation, large numbers of individuals would require biological dosimetry • BUT: • Immediate medical care based on symptoms not dose • Accurate dose estimates of lesser importance • examining 50 spreads should be sufficient for sorting of casualties (1 hour scoring/sample) • Slides can even be prioritized after scoring 20 spreads • More accurate dose estimates can be made later by counting more spreads

9. Normal Other strategies for increasing biodosimetry throughput • QuickScan • With DCA, each cell must have 46 centromeres (ie must be scored) • With QuickScan, don’t score centromeres - only identify damage • Greatly decreases scoring time • Requires scorer to experience to identify complete spreads (Health Physics, 2009, In Press) • Further validation underway

10. 1.4 HC 1.2 DRDC AECL 1.0 0.8 MN/BN cell 0.6 0.4 0.2 0.0 0 1 2 3 4 Dose (Gy) Other strategies for increasing biodosimetry throughput • Cytokinesis Block Micronucleus Assay • Count micronuclei in binucleated lymphocyte cells • Faster to score • Easier to train scorers • Method validated for NBDRP in 2008 • Each lab needs its own reference curve • All scorers within each lab are similar • All scorers in each lab could detect (statistically) unknown doses ≥ 1 Gy, after scoring only 200 BN cells (Radiation Protection Dosimetry, 2009, In Press)

11. Increasing number of trained personnel through networking, teaching and exercises Reference laboratories Satellite laboratories 18 clinical laboratories across the country

12. REFERENCE LABORATORIES • 4 Canadian CRTI partners (reference laboratories) now have enhanced capacity to conduct the DCA analysis • The role of these 4 reference labs include • Developing SOP and working standards for triage DCA • Generating of reference curves for different radiation qualities • Maintaining expertise and capacity through annual exercises • Providing advice and assistance following R/N event

13. CORE LAB EXERCISES • Two exercises completed during this project. • 10 blood samples/lab processed • 1st exercise, December 2007, 4 Canadian labs (15 scorers) • 2nd exercise, November 2008, 4 Canadian labs + 2 US labs (19 scorers)

14. SUMMARY OF EXERCISE RESULTS • Within a week, the equivalent of 180 samples were scored • Well below each lab’s capacity. • from EX07 to EX08: -increased # of trained scorers, especially in Quick Scan -increased accuracy with all methods -decreased scoring time for DCA • Estimated capacity 200-300 samples/week (Canadian core labs alone)

15. TRAINING PROGRAM FOR CYTOGENETICISTS • Developed a one day training course delivered to first year cytogenetics students at the Michener Institute (16 students/year) • Two sessions given so far (Nov 2007 and 2008) • Radiation lecture • DNA strand break syndrome lecture • Dicentric Assay training with students scoring • NEW this year – Blood processing lab

16. COLLABORATIVE AGREEMENT WITH MICHENER • In addition, Collaborative Agreement with Michener has been signed • To use their large resources a Laboratory Centre for Emergency Response for processing and scoring samples in a central location • In return, provide them with training, equipment and reagents. • Will increase Canadian capacity to ~1000/week

17. STRENGTHEN LINKS WITH CLINICAL LABORATORIES • Identify up to 5 clinical laboratories for training for more formal arrangements for biological dosimetry • Deliver information sessions and on-site training to selected clinical laboratories • Establish stronger links with the selected clinical laboratories through discussions with each of the laboratories on how agreements can be established • Discussion commenced with two laboratories • Proposing $5-8K/year incentive (contract) • First meeting with labs on June 30th • Conduct exercises involving these laboratories • Increases Canadian capacity to additional ~50/week/laboratory

18. INTERNATIONAL ACTIVITIES: • International Networking: • WHO 1st Consultation on the Development of a Global Biodosimetry Laboratories Network for Radiation Emergencies (Dec 2007) • Planning started for developing an international biological dosimetry response network • Meeting report published in Radiation Research (Rad. Res., 171, 2009) • Canada suggested as one of the reference laboratories • Links Canada into a huge resource for biological dosimetry capacity

19. INTERNATIONAL ACTIVITIES: • International Atomic Energy Agency • Working towards registering capabilities with the Response Assistance Network (RANET) • Assisting in revisions of Technical Report Series #405 Cytogenetic analysis for Radiation dose assessment • To be published 2010

20. PROJECT SUMMARY AND CONCLUSIONS • NBDRP is becoming well established and recognized around the world • Capacity at core labs has been increased (16 trained individuals) • Ready to respond to emergency • Capacity will continue to increase with: • continued training and agreements with Michener Institute • Agreements with cytogenetic laboratories • Strengthening links with US laboratories • Involvement with WHO and IAEA international networks • New high throughput methods for biological dosimetry

21. FUTURE DIRECTIONS • Continue with annual exercises and developing international networks • Continue research on: • Biomarkers for radiation response • Interphase micronucleus assay by flow cytometry • Other arising novel methods for biodosimetry • Plan workshop “Biodosimetry- present status for emergency response in North America” • Invite US partners • 1-2 days of talks • Bring in policy makers to talk about Canada/US links (ie CRTI, NIH)

22. ACKNOWLEDGEMENTS Health Canada Catherine Ferrarotto Barbara Kutzner Sylvie Lachapelle Nancy Ringuette Dr. James McNamee Pascale Bellier DRDC Hillary Boulay Greene Louise Prud’homme-Lalonde McMaster University Dr. Doug Boreham Dr. Jo-Anna Dolling AECL Farrah Flegal Dale Buchanan Yvonne Devantier Laura Patterson Armed Forces Radiobiology Research Institute Dr. Pat Prasanna Oak Ridge Institute for Science and Education REAC/TS Dr. Gordon Livingston