AFRRI
A Unique National Resource
Who We Are and What We Do
Michael P. Dempsey, PhD, MT(ASCP)
Major, USAF, BSC
SAFMLS 2010
San Diego, CA
25 MARCH 2010
Presentation Objectives
• 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
Established 1960
Background
• 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.
The Threat
• Accidents
• Radiological
dispersal devices
• Radiation-emitting
devices
• Nuclear weapons
AFRRI Mission
Medical 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
Medical Consequences of
Acute Radiation Injury
Gastrointestinal (5-20 Gy)
Hematopoietic
(1-6 Gy)
Cardiovascular
CNS
(>20 Gy)
Subclinical
Increasing Dose
Detonation Casualties
1 KT
10 KT
>7k
>13k
~18k
~114k
Intensive care (IC) ward (530-830): ~19k
~90k
Prompt fatalities:
Expectant (>830 cSv):
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 radiationinduced hematopoietic or GI injury
Scientific Program
Prophylaxis
Therapy
Protection
Assessment
Treatment
• Candidate screening
• Biodosimetry:
Cytogenetics and
Molecular markers
• Drug evaluation
• Drug evaluation
• Automation
S
U
R
V
I
V
A
L
• Combined effects
• Heavy metal toxicity
Scientific Functional Schematic
Scientific Director
Scientific
Research
Department
Radiation
Countermeasures
Biological
Dosimetry
Veterinary
Sciences
Department
Radiation
Sciences
Department
Radiation
Neutralization
Research
Support
Group
Internal
Contamination
& Metal Toxicity
Radiation
Combined Injury
Radiation Sciences Department
Unique Radiation Facilities
1 Megawatt TRIGA Mark F Reactor
Phillips Industrial X-Ray
Facility
100 Curie Cesium
Calibration Facility
Chronic Irradiation
Facility
450,000 Curie Co-60
Panoramic Irradiator
450 K curies
Provides researchers large,
uniform gamma-ray fields
with a wide array of
exposure configurations
Veterinary Sciences Department
Resources
• 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 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
Research Support Group
AFRRI FY09 Research Funding
The Research Support Group (RSG)
• Formed per recommendations from the Scientific
Directorate (SD) strategic planning team.
• The transformation to a principal investigatorcentric 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.
FY09
Intramural
$2,146,000
FY09
Extramural
$9,028,254
Some of AFRRI’s Extramural Sponsors
NIAID, DTRA, NIH, NASA, DARPA, BARDA
Scientific Program Areas
Biological Dosimetry
Program
William F. Blakely, PhD
Program Advisor
Mission: To develop rapid, high-precision methods that
determine radiation exposure of radiological casualties for
use in triage and medical management addressing:
• 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.
Website for research program:
http://www.afrri.usuhs.mil/research/biodos.htm
Website 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 masscasualty 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, PhD
Program 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 TPO Mimetic Enhances Survival in Mice B Intracellular Signaling Pathways: DT3
DT3
AKT
Survival (%)
ERK
X
P
4EBP
eIF4E
S6K
S6
Protein translation
Cell survival and growth
Time Post-Irradiation (Days)
C
mTOR
D
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 DeltaTocotrienol (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).
Recent Accomplishments
• 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
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 gammaradiation 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, PhD
Program 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. Survival
(%)
Survival
% Survival
120
100
SHAM
Wound
80
8.5 Gy
60
8.5 Gy+Wound
40
9.75 Gy
9.75 Gy+Wound
20
0
0
5
10
15
20
25
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%
30
Days after RI (Days)
or CI
Postirradiation
C. CI
IL-6 (pg/ml)
B. Wound
D. IL-6
* *
**
Postirradiation (Days)
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 longterm 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 highthroughput 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
A
B
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.
Selected Recent Publications
• 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)
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
Operational Support
Training
Medical 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)
AFRRI CIVILIAN PERSONNEL SUMMARY
PAYGRADE TYPES
Current
Onboard*
Civilian Federal
(t=66)
GS
50
WG/WL
16
Contractors
HJF
(t=62)
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 Billets
FY 09
Onboard*
% Manning
Officers
17
15
88%
Enlisted
13
11
85%
Officers
7
5
71%
Enlisted
11
11
100%
Officers
9
3
33%
Enlisted
7
5
71%
64
50
75%
Civilian
Military
128
50
Army
Navy
Air Force
TOTAL
Personnel Type
Personnel Totals
*Onboard totals as of 3/1/10
178
Types of Military and Civilian Positions
Position Titles
Civilian
Military
RESEARCH BIOCHEMIST
x
x
RESEARCH BIOLOGIST
x
x
RESEARCH MICROBIOLOGIST
x
x
RESEARCH CHEMIST
x
RESEARCH PHYSICIST
x
RESEARCH PHYSIOLOGIST
x
BIO STATISTICIAN
x
BIOLOGICAL SCIENCE TECH
x
x
HEALTH PHYSICIST
x
x
HISTOTECHNOLOGIST
(COMPARATIVE PATHOLOGY)
x
NUCLEAR ENGINEER
x
x
ADVANCED LAB TECH
x
x
VETERINARIAN
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
Acknowledgements
COL Patricia Lillis-Hearne
COL Donald Hall
Dr. Christopher Lissner
CDR John Gilstad
MAJ Larry Shelton
Mr. Steve Miller
LT Matthew Deshazo
Director
Deputy Director
Interim Scientific Director
Head, Scientific Research Dept.
Head, Veterinary Science Dept.
Head, Radiation Science Dept.
Head, Admin Support Dept.
SAFMLS Presentation Team:
Maj Michael Dempsey
CPT Nabil Latif
CPT Anthony Kang
HM1 Neal Agravante
HM2 Sergio Gallego
For more information go to: www.afrri.usuhs.mil
AFRRI STAFF