AFRRIA Unique National Resource Who

We Are and What We Do

AFRRIA Unique National Resource Who

We Are and What We Do

Michael P. Dempsey, PhD, MT(ASCP)Major, USAF, BSC

SAFMLS 2010San Diego, CA

25 MARCH 2010

Michael P. Dempsey, PhD, MT(ASCP)Major, USAF, BSC

SAFMLS 2010San Diego, CA

25 MARCH 2010

• Provide introduction to AFRRI’s history and mission

• Introduce on-going scientific research and education at AFRRI

• Describe AFRRI assignment opportunities for DoD laboratory scientists

Presentation Objectives

Established 1960

• AFRRI is the only medical nuclear/radiological defense Research and Development Institute in DoD.

• We are strategically aligned with the Uniformed Services University of Health Sciences.

Background

The Threat

• Accidents

• Radiological dispersal devices

• Radiation-emitting devices

• Nuclear weapons

AFRRI MissionMedical R&D • Conduct radiobiology research and develop

medical countermeasures for DoD.

Medical Education • Train medical personnel in ionizing radiation

countermeasures.

Medical Emergency Response Team • Advise JCS (J-4 Medical), Combatant

Commands, and others on radiological matters.

Consultation• Answer questions from federal agencies and

participate with them as subject matter experts.

AFRRI Research in Radiation Biology

Hematopoietic(1-6 Gy)

Gastrointestinal (5-20 Gy)

Cardio-vascular

Subclinical

CNS(>20 Gy)

Increasing Dose

Medical Consequences ofAcute Radiation Injury

Detonation Casualties

1 KT 10 KT

Prompt fatalities: >7k >13k

Expectant (>830 cSv): ~18k ~114k

Intensive care (IC) ward (530-830): ~19k ~90k

IC/minimum care ward (300-530): ~33k ~141k

Minimum care ward (150-300): ~66k ~150k

Outpatient (70-150): ~83k ~159k

Health monitoring (25-70): ~106k ~128k

Worried well (<25): >150k >212k

Research Gaps in Medical Preparedness

• Countermeasures– The only FDA-approved radioprotectant, amifostine, has toxicity that

makes military use not feasible

• Biodosimetry– Biodosimetric tools for triage are limited in speed and physiologic

predictive power

• Mitigation/Therapy– No drug has an FDA-approved indication for radiation-induced

hematopoietic or GI injury

Prophylaxis

SURVIVAL

Assessment• Biodosimetry: Cytogenetics and Molecular markers

• Automation

Protection• Candidate screening

• Drug evaluation

Treatment• Drug evaluation

• Combined effects

• Heavy metal toxicity

Scientific Program

Therapy

Biological Dosimetry

Radiation Neutralization

Internal Contamination

& Metal Toxicity

RadiationCountermeasures

RadiationCombined Injury

Radiation Sciences

Department

Research Support Group

Veterinary Sciences

Department

Scientific Director

Scientific Research

Department

Scientific Functional Schematic

Radiation Sciences Department

Unique Radiation Facilities

1 Megawatt TRIGA Mark F Reactor

450,000 Curie Co-60 Panoramic Irradiator

Chronic Irradiation Facility

Phillips Industrial X-Ray Facility

100 Curie Cesium Calibration Facility

Provides researchers large,uniform gamma-ray fieldswith a wide array ofexposure configurations

450 K curies

• Animal facility designed to support radiation & surgery studies (32K ft2)

• State-of-the-art environmental controls & monitoring

• Histopathology, Microbiology, and Clinical Pathology Labs

• AAALAC-accredited animal care and use program

• Supports AFRRI, SOM, WRAMC, and Navy studies

• All protocols IACUC approved

Veterinary Sciences DepartmentResources

Veterinary Sciences Department

Staffing• Currently 27 Staff Members

• 4 Veterinarians: Department Head, Laboratory Animal

Medicine Resident, Contract Clinical Veterinarian, and

Veterinary Pathologist

• 5 Veterinary Technicians: 3 Army and 2 Civilian

• 12 Government Husbandry Personnel

• 3 Pathology Lab Staff (2 GS; 1 Air Force)

• 3 Administrative Personnel

Some of AFRRI’s Extramural Sponsors NIAID, DTRA, NIH, NASA, DARPA, BARDA

Research Support Group

The Research Support Group (RSG) •Formed per recommendations from the Scientific Directorate (SD) strategic planning team. •The transformation to a principal investigator-centric organization allows RSG to provide support to our scientists and minimize the growing demands that keep them out of the laboratory.

RSG staff (5 employees)•Cooperateve Research and Development Agreements (CRDA)•Intramural/extramural proposals (pre & post award), budgets, and Gov’t credit cards. •Works with contracts, equipment/service agreements as well as Good Laboratory Practices (GLP) development.

Scientific Program Areas

Biological Dosimetry Program

William F. Blakely, PhDProgram Advisor

• Automation and field deployability for rapid battlefield dose assessment•Enhancement of reference biodosimetry capability• Identification and validation of radiation specific biomarkers for early phase, organ-specific, and late effects.

Mission: To develop rapid, high-precision methods that determine radiation exposure of radiological casualties for use in triage and medical management addressing:

Website for research program: http://www.afrri.usuhs.mil/research/biodos.htmWebsite for biodosimetry tools: http://www.afrri.usuhs.mil/outreach/biodostools.htm

Focus Areas and Staffing• Identification, optimization, and validation of integrated biodosimetry

assays for military and civilian applications with the following emphases:

• Development of medical recording and software-based applications to manage radiation dose and injury assessment

• Characterization of prodromal clinical signs and symptoms for radiation exposure diagnostics

• Automation of cytogenetic assays for radiation dose assessment

• Identification of radiation bioassays using i) nucleic-acid based methodology and ii) for early-expressed and persistent radiation late effects

• Combined use of hematology biomarkers with clinical signs/symptoms for radiation dose assessment

• Validation of blood proteomic, metabeolomic, and urinary biomarkers for early-phase and organ-specific radiation injury and dose assessment

• Integration of multi-parameter early-phase clinical biodosimetry

• Staff: 3 primary and 3 associated PIs, 1 doctoral level Investigator, 2 Programmers, 2 Postdocs, 2 masters level Research Associates, 2 primary and 2 associated Laboratory Technicians, 2 volunteers (Total: 19)

Recent Accomplishments• Patents and IDE

• U.S. patent filed (March 2009); international patent filed (October 2009)

• Pre-IDE meeting with FDA (regulation of automated sample processing for cytogenetic dose assessment)

• Other• Medical Communications for Combat Casualty Care (MC4) deployed AFRRI’s BAT

software application to CONUS and OCONUS (see website: www.mc4.army.mil).

• In 2010, 3 of 9 BARDA contract awardees collaborating with AFRRI scientists on a radiation-responsive proteomic diagnostic concept for rapid & early-phase biodosimetry device(s).

• Participation in international cytogenetic biodosimetry inter-comparison study and the development of a cytogenetic network concept to enhance sample analysis from a mass-casualty radiological incident.

• In 2009 AFRRI BD scientists established a nonhuman-primate radiation dose-response model at AFRRI for the purpose of validating candidate novel and multiple biodosimetry assays, critical for obtaining necessary FDA approvals.

• Additional products and accomplishments of the AFRRI’s BD Research Group are available at website: www.afrri.usuhs.mil/research/biodos.htm

Selected Recent Publications• Ossetrova NI et al. (2010) Combined approach of hematological biomarkers and plasma protein

SAA for improvement of radiation dose assessment in triage biodosimetry applications. Health Physics 98(2):204–8.

• Prasanna PGS et al. (2010). Triage dose assessment for partial-body exposure: Dicentric analysis. Health Physics 98(2):244–51.

• Blakely WF, Ossetrova NI et al. (2010) Multiple parameter radiation injury assessment using a nonhuman primate radiation model—biodosimetry applications. Health Physics 98(2):153–9.

• Prasanna PGS, Blakely WF et al. (2010) Synopsis of partial-body radiation diagnostic biomarkers and medical management of radiation injury workshop. Radiation Research 173(2):245–53.

• Ossetrova NI, Blakely WF (2009) Multiple blood-proteins approach for early-response exposure assessment using an in vivo murine radiation model. International Journal of Radiation Biology 85(10):837–850.

• Blakely WF, Carr Z et al. (2009) WHO 1st Consultation on the Development of a Global Biodosimetry Laboratories Network for Radiation Emergencies (BioDoseNet). Radiation Research 171(1):127–139.

• Waller E, Millage K, Blakely WF et al. (2009) Overview of hazard assessment and emergency planning software of use to RN first responders. Health Physics, 97(2):145–156.

• Pathak R, Ramakumar A, Subramanian U, Prasanna PG. (2009) Differential radio-sensitivities of human chromosomes 1 and 2 in one donor in interphase- and metaphase-spreads after 60Co gamma-irradiation. BMC Medical Physics 9:6 8-pages (open access journal, website: www.biomedcentral.com/1756-6649/9/6).

Radiation Countermeasures Program

Mark H Whitnall, PhDProgram Advisor

Mission: To develop pharmacological countermeasures to radiation injury that can be used by• Military personnel and • Emergency responders

Focus Areas and Staffing• Developing radiation countermeasure drugs in 8 individual research

programs with the following emphases:• Gamma-Tocotrienol, Intracellular Signaling, Screening Drugs with In Vivo Mouse Model

• Ex-Rad, Gamma-Tocotrienol, Intracellular Signaling

• Genistein, BIO300, In Vivo Hematopoiesis

• Cancer prevention, Intracellular Signaling

• Alpha-Tocopherol Succinate, CBLB502, Myeloid Progenitor Cells, Cytokine Expression

• Delta-Tocotrienol, SOD Mimetic, TPO Mimetic, Screening Drugs with In Vivo Mouse Model, Mitochondrial Metabolism

• In Vitro Hematopoietic Niche Model, Intracellular Signaling, Neutron/Gamma Fields, Minipig Model for Drug Development

• In Vitro Hematopoietic Niche Model, Delta-Tocotrienol, Intracellular Signaling, In Vivo Hematopoiesis

• Staff: 8 PIs, 11 Doctoral Level Investigators, 1 Masters Level Investigator, 25 Laboratory Technicians, 1 Intern, 2 Administrative (Total: 48)

A: 30 day mouse survival after whole-body gamma-irradiation: Effect of a thrombopoietin (TPO) mimetic (sc, 24 h before irradiation). B: Intracellular signaling pathways modulated by Delta-Tocotrienol (DT3). C: Mouse bone marrow, 8 days after whole-body gamma irradiation (7 Gy). D: As in C, but mouse treated with Alpha-Tocopherol Succinate (sc, 24 h before irradiation).

A B

C D

TPO Mimetic Enhances Survival in Mice

Su

rviv

al (

%)

Time Post-Irradiation (Days)

Intracellular Signaling Pathways: DT3

AKTERK

S6K

S6

Protein translationCell survival and growth

4EBPP

mTOR

eIF4E

Radiation

DT3

X

• Patent approval: Landauer et al., “Phytoestrogenic isoflavone compositions … for protection against and treatment of radiation injury”

• Other• Developed minipig model for advanced radiation

countermeasure development• Integrated mechanistic and applied research with advanced drug

development• Working with companies to pursue advanced development of

countermeasure candidates (large animal trials, attend FDA meetings, etc.)

• All four radiation countermeasures with FDA IND status are AFRRI products

Recent Accomplishments

Selected Recent Publications

• Merlot R, Huang G, Houpert H, Miller AC, Lison P. The Inhibitory Action of Epigallocatechin gallate, a DNA Methyltransferase Inhibitor, on Neoplastic Transformation. Anti-Cancer Research. Epub Mar 1, 2010

• Singh VK, Brown DS, Kao TC: Alpha-tocopherol succinate protects mice from gamma-radiation by induction of granulocyte-colony stimulating factor. Int J Rad Biol. 86: 12-21, 2010.

• Ghosh SP, Kulkarni S, Hieber K, Toles R, Romanyukha L, Kao TC, Hauer-Jensen M, Kumar KS. Gamma-tocotrienol, a tocol antioxidant as a potent radioprotector. Int J Radiat Biol 85: 598-606, 2009.

• Singh VK, Grace MB, Parekh VI, Whitnall MH, Landauer MR: Effects of genistein administration on cytokine induction in whole-body gamma irradiated mice. Int Immunopharmacol. 9: 1401-1410, 2009.

• Xiao M, Inal CE, Parekh VI, Li XH, Whitnall MH. Role of NF-kappaB in hematopoietic niche function of osteoblasts after radiation injury. Exp Hematol 37: 52-64, 2009.

• Srinivasan V, Doctrow S, Singh VK, Whitnall MH. Evaluation of EUK-189, a synthetic superoxide dismutase/catalase mimetic as a radiation countermeasure. Immunopharmacol Immunotoxicol 30: 271-290, 2008.

Radiation Combined Injury Program

Juliann G. Kiang, PhDProgram Advisor

Mission: To develop medical treatments for irradiated personnel whose exposure is compounded by traumatic

wounds, burns, hemorrhage, and/or infection. Treatment strategies under investigation include biological response modifiers, new antimicrobial

agents, probiotics, and stem cells, used individually or in combination.

Focus Areas and Staffing

• Developing radiation countermeasure drugs in 6 individual research programs with the following emphases:• 17-DMAG, Intracellular Signaling, Screening Drugs with Mouse Model and In

Vitro Model

• ARA290, S-TDCM combined with levofloxacin, and Bone Marrow Mesenchymal Stem Cells, Screening Drugs with Mouse Model

• A Novel In Vivo Combined Injury Model to be established

• Levofloxacin, Pharmacodynamics and Pharmacokinetics

• Staff: 3 PIs, 3 military officers, 5 Doctoral Level Investigators, 1 Master Level Investigator, 6 Laboratory Technicians, 3 Interns (Total: 21)

Recent Accomplishments• Two patents and one MTA

A. B6D2F1/J mice received 8.5 or 9.75 Gy Co-60 gamma irradiation followed by 15% total body surface area wounding. The survival in % 30 days after irradiation or irradiation+wounding: Sham -100%; Wound - 100%; 8.5 Gy - 100%; 8.5 Gy+Wounding -70%; 9.75 Gy – 60%; 9.75 Gy+wounding - 10%

0 5 10 15 20 25 30

0

20

40

60

80

100

120SHAMWound

9.75 Gy

9.75 Gy+Wound

8.5 Gy

8.5 Gy+Wound

Days after RI or CI

Su

rviv

al (%

)

B. Wound C. CI

D. IL-6

IL-6

(p

g/m

l)

Postirradiation (Days)

*** *

A. Survival

Postirradiation (Days)

% S

urv

ival

B6D2F1 mice received 9.75 Gy Co-60 gamma irradiation followed by body surface wounding. B. A good size of the healing bud in wounded area. C. Irradiation reduces the healing bud in wounded area, causes degeneration of adipocytes, and eliminates neutrophils and macrophages. D. Wounding enhances irradiation-induced increases in IL-6 levels.

Selected Recent Publications• Kiang JG, Garrison BR, Gorbunov NV. Radiation combined injury: DNA damage,

apoptosis, and autophagy. Adapt Med 2: 000-000, 2010 (in press).

• Kiang JG, Jiao W, Cary L, Mog SR, Elliott TB, Pellmar TC, Ledney GD. Wound trauma increases radiation-induced mortality by increasing iNOS, cytokine concentrations, and bacterial infections. Radiate Res 173: 000-000, 2010 (in press).

• Kiang JG, Smith JA, and Agravante NG. Geldanamycin analog 17-DMAG inhibits iNOS and caspases in gamma irradiated human T cells. Radiat Res 172: 321-330, 2009.

• Gorbunov NV, Kiang JG. Up-regulation of Autophagy in the Small Intestine Paneth Cell in Response to Total-Body γ-Irradiation. J Pathol 219: 242-252, 2009.

• Jiao W, Kiang JG, Cary L, Elliott TB, Pellmar TC, Ledney GD. COX-2 inhibitors are contraindicated for therapy of combined injury. Radiat Res 172: 686-697, 2009.

• Ledney GD, Elliott TB. Combined injury: factors with potential to impact radiation dose assessments. Health Phys 98:145-152, 2009.

Internal Contamination and Metal Toxicity Program

John F. Kalinich, Ph.D.

Program Advisor

Mission: To determine whether the short-term and long-term radiological and toxicological risks of embedded metals warrant changes in the current combat and postcombat fragment removal policies for military personnel and, in the case of internalized radiological hazards, investigate treatment strategies to enhance elimination of these metals from the body.

Focus Areas and Staffing

• Health effect of embedded metal fragments, including DU and tungsten alloy, with a focus on carcinogenicity and transgenerational effects.

• Identification of biomarkers of exposure to internalized radionuclides and development of high-throughput analytical systems.

• Decorporation procedures for the enhanced elimination of radionuclides from the body as a result of a nuclear explosion or “dirty bomb” event.

Staff: 2 PIs, 2 Research Associates, 2 Lab Technicians

Photomicrograph of J774 cells untreated (Panel A) or treated with depleted uranium (Panel B) for 24 h then stained with 2-(5-bromo-2-pyridylazo)-5-diethylaminophenol to indicate uranium deposits.

A B

• Military Medicine 174: 265-269 (2009) • Biochemie 91: 1328-1330 (2009) • Toxicology In Vitro 23: 356-359 (2009) • Military Medicine 173: 754-758 (2008) • Radiation Measurements 42: 1029-1032 (2007) • Reviews on Environmental Health 22: 75-89 (2007)• Environmental Health Perspectives 113: 729-734 (2005)• Molecular and Cellular Biochemistry 279: 97-104 (2005)

Selected Recent Publications

Radiobiology Education and Training Opportunities

• USU Department of Radiation Biology – PhD program coming

• PhD in Molecular and Cellular Biology: Radiation Biology track

• Operational Support Training: MEIR

Medical Effects of Ionizing Radiation CourseMedical Effects of Ionizing Radiation Course

• CME / CNE /CHE credit

• Required training for contingency personnel

• Target Audience

• Physicians

• Nurses

• Medical Response Personnel

• 30 - 40 courses presented worldwide annually with approximately 1000 individuals trained

• Courses are customizable (1-day focused courses)

Operational Support Training

AFRRI CIVILIAN PERSONNEL SUMMARY

PAYGRADE TYPES Current Onboard*

Civilian Federal (t=66)

GS 50

WG/WL 16

Contractors (t=62)

HJF 48**

Corbin 2

JCS 2

Eagle Security 10

TOTAL 128

*Onboard totals as of 3/1/10**30 under contract, 18 under grants

AFRRI Authorized Military BilletsFY 09 Onboard* % Manning

Army

Officers 17 15 88%

Enlisted 13 11 85%

Navy

Officers 7 5 71%

Enlisted 11 11 100%

Air Force

Officers 9 3 33%

Enlisted 7 5 71%

TOTAL 64 50 75%

*Onboard totals as of 3/1/10

Personnel Type Civilian Military

Personnel Totals 128 50 178

Types of Military and Civilian Positions

Position Titles Civilian MilitaryRESEARCH BIOCHEMIST x xRESEARCH BIOLOGIST

x xRESEARCH MICROBIOLOGIST

x xRESEARCH CHEMIST

xRESEARCH PHYSICIST

xRESEARCH PHYSIOLOGIST

xBIO STATISTICIAN

xBIOLOGICAL SCIENCE TECH

x xHEALTH PHYSICIST

x xHISTOTECHNOLOGIST (COMPARATIVE

PATHOLOGY) xNUCLEAR ENGINEER

x xADVANCED LAB TECH

x xVETERINARIAN x x

VETERINARIAN TECH x x

Perspectives of an AFRRI Enlisted Military Lab

Technician

HM2(SW) Sergio Gallego

AFFRI’s Opportunities

Opportunity to contribute to the DoD’s effort in finding means to counter the effects of ionizing radiation.

Be part of developing new protocol/testing procedures to be used in the future, out in the field environment or in combat setting.

Learn how to perform research and understand the different stages of research while working with world renowned scientists.

AFRRI’s Opportunities (con’t)

• Learn new laboratory procedures and operate highly sophisticated equipment not normally available in basic clinical lab setting:

- Protein assays / Western Blots

- PCR/Molecular biology procedure – Bio Rad IQ5

- Cell counting / Flow Cytometry – Guava 8HT

- Automated RNA isolation & purification – Qiagen Bio Robot 8000

- Immunohistochemical staining

- Stem Cell Isolation and culture (bone marrow)

AFRRI’s Uniqueness

• Perform various veterinary procedures such as obtaining tissue/fluid samples and administering therapeutics.

• Perform animal survival studies and irradiate experimental animals using different radiation sources.

• See the stages of your work as it progresses from start to end.

Operations Bushmaster and Kerkesner

• This field exercise is unique to USU's medical school curriculum and is designed to ensure that USU graduates are well prepared to practice "good medicine in bad places."

• Students are trained to deal with wartime casualties, national disasters, emerging infectious diseases and other public health emergencies.

Why AFRRI?

• Tri-Service Experience

• Joint Service/DoD Awards

• Continuing Education Opportunities

• USUHS Affiliation

• WRNMMC as MHS epicenter in 2011

• DC and surrounding area, including NIH

COL Patricia Lillis-Hearne DirectorCOL Donald Hall Deputy DirectorDr. Christopher Lissner Interim Scientific DirectorCDR John Gilstad Head, Scientific Research Dept.MAJ Larry Shelton Head, Veterinary Science

Dept.Mr. Steve Miller Head, Radiation Science Dept.LT Matthew Deshazo Head, Admin Support Dept.

SAFMLS Presentation Team:Maj Michael DempseyCPT Nabil LatifCPT Anthony KangHM1 Neal AgravanteHM2 Sergio Gallego

For more information go to: www.afrri.usuhs.mil

Acknowledgements

AFRRI STAFF