From Spirochetes to Select
Agents: Thoughts on an
Ongoing Science Career
Andrea Keane-Myers, Ph.D.
Vaccines and Medical Countermeasures
Biological Defense Research Directorate
Naval Medical Research Center
Overview
• What have I done?
• Why did I make these career choices?
• What am I currently doing?
• Job market/ funding
• What are my trainees are currently doing?alternative career paths
• Work/Life Balance
What have I done?
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School
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Post Doctoral Fellowships
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Post Doc- Pulmonary Physiology (JHMI)
Post Doc- Ocular Immunology (Harvard)
Junior Faculty
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B.S. in Biology/ Studio Art (University of Richmond)
Ph.D. in Molecular Microbiology and Immunology (Johns Hopkins Bloomberg School of
Public Health)
Instructor- Ocular Immunology (Harvard)
Staff Scientist- Laboratory of Immunology (NEI/ NIH)
“Real Jobs”
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Chief, Allergic Inflammation Section (NIAID/NIH)
Head, Vaccines and Medical Countermeasures (BDRD/ NMRC)
School-
What do I want to be when I grow up?
B.S.
Biology/ Studio Art
(University of Richmond)
M.D.?
Marine Biology?
Hives*  Immunology Research
Spicule formation- SEM
Ph.D.
Molecular Microbiology and Immunology
Johns Hopkins Bloomberg School of Public
Health
Lyme Disease*
Medical Illustration?
Borrelia burgdorferi infection
* Personal experience
Post Docs-
90 degrees from previous training
Pulmonary Physiology/
Immunology
Johns Hopkins Medical Intuitions
Asthma*
Pulmonary
function testing
Ocular/ Molecular Immunology
Schepens Eye Research
Institute/ Harvard
Allergic Conjunctivitis*
Mucosal response
Transgenic mice
* Personal experience
Junior FacultyWorking towards finding a “Real Job”
Instructor
Ocular Immunology
Schepens Eye Research Institute/ Harvard
T cell Tetramer
Allergic Conjunctivitis*
Cat Dander Response*
Staff Scientist
Laboratory of Immunology
NEI/NIAID
Clinical Allergic Conjunctivitis*
Clinical allergic eye disease
Cat dander- Fel d1
T cell tetramer
* Personal experience
“Real Jobs- NIH”
Chief, Allergic Inflammation Section
NIAID/NIH
Asthma*
Conjunctivitis*
Parasite Infection- Ascaris #
PBS
A. suum-infected
*Personal experience
# Diseases I haven’t had (and don’t want)
Helminth Infection and Allergic Disease
Allergy
Th2
Ascaris
infection
Th2
Ascaris infects 1.5 billion people a year (#1 geohelminth)
Ascaris epidemiology: more parasites less allergic disease
Can Ascaris products induce regulation of inflammation?
Can we exploit the Ascaris regulatory components and develop
therapeutics to prevent allergic inflammation?
2) Receptor?
3) Cellular activation/
signal transduction
MHC
CD86
CD80
TLR4
(other PRR?)
DC
NF-kB
ERK
CD40
IL-10
1) Biochemical
analysis
PCF
IL-12
ABA-1
x
heme
-binding
Glycolipids
CD4+
T effector
cell
IL-4
IL-5
IL-13
IL-10
6) Use PCF components
therapeutically
5) T cell migration
CD25T reg cell
CD25+
T reg cell
“Real Jobs- BDRD/NMRC”
Head, Vaccines and Medical Countermeasures
Biologic Defense Research Directorate
Naval Medical Research Center
Vaccines for Bacterial Select Agents ##
Job: To design
vaccines and
therapeutics to
protect the
Warfighter from
bacterial select
agents so they can
complete their
mission.
Bacillus anthracis
Yersinia pestis
Francieslla tularensis
Burkholderia pseudomallei
## Diseases I haven’t had (and really, really don’t want)
BDRD Vaccines and Medical Countermeasures
Target discovery
Therapeutics
Rapid vaccine delivery systems
Inhaled and oral delivery
Nanoparticles (OSU)
Spores
Biology
Identification of Novel Pathogen targets
Iron regulated Membrane Proteins (Sidephores- Syntiron)
Conserved Genes from Intracellular Pathogens
T and B cell epitopes
DNA-based vaccines and therapies
Pre-exposure protection – human antibodies (IQ)
Electroporation (Ichor)
Human monoclonal antibodies
Post-exposure protection
Genomics
Innate and adaptive immune agonists/adjuvants
Innate immune modulators
TLR agonists, PAMP targets
Innovative Vaccine Protein Production
Caterpillars (Chesapeake Perl)
Tobacco Plants (Fraunhofer)
In house protein production- saves $ millions
Immunology
Development of animal models of human disease
BDRD Vaccines and Medical
Countermeasures
 30+ years experience in immunology, microbiology
 Access to select agents (including a large library of
environmental isolates)
 BSL-2 and BSL-3 level laboratories and animal
facilities (IACUC-approved protocols)
 Established rodent models for in vivo challenge with
select agents
 Internal production of recombinant microbial
proteins has saved DoD 5+ million dollars
 Collaboration with Navy Medical Health community
for transition of products to the clinic
For Official Use Only- Not for Distribution
Projects and Partnerships
• BDRD (Vaccine and Genomics Departments)- miRNA
analysis post select agent exposure
• Syntiron- Iron Acquisition Receptor Vaccines
• Ohio State- Nanoparticle Vaccine Delivery Platforms
• Epivax-T cell Epitope Based Vaccines
• Ichor- Electroporation Delivery of DNA Vaccine
Platforms
• Biological Mimetics Incorporated- Immune refocusing
technology
•
The Vaccines and Medical Countermeasures Group is always looking for new
Teaming possibilities
For Official Use Only- Not for Distribution
Ac-DEX microparticles encapsulating
bacterial lysate
Scanning electron micrograph of representative Ac-DEX microparticles encapsulating F.
novicida lysate. Scale bar is 10 microns.
Particle Set
F. novicida Lysate
Resiquimod
Wt loading (mg/100mg AcDEX)
1.70 ± 0.14
0.22 ± 0.00037
%Efficiency
47.3 ± 3.8
7.4 ± 0.012
DARPA Live Fire Exercise-
Protection against an unknown select agent in 7 days
0
40
0
Prime
7
Boost
14
1st
Challenge
1.49x106 CFUs
For Official Use Only- Not for Distribution
Live Fire Exercise
Survival Curves- B. pseudomallei
Figure 4: Survival of mice (n=25) vaccinated with various formulations (day -14, -7) and then challenged ip with
live fire agent. NP = nano/microparticle; Protein = live fire lysate; Resiq = resiquimod; /NP = encapsulated in AcDEX microparticle.
For Official Use Only- Not for Distribution
Electroporation with Ichor’s TriGrid Array
Electroporation with DNA Protects Mice from Anthrax
Challenge as well as recombinant PA vaccination
*Recombinant protein vaccinations were
performed with 10 µg of protein per
mouse.
Combination DNA vaccines protect mice from
plague challenge
*Recombinant protein vaccinations were
performed with 10 µg of protein per
mouse.
BDRD Vaccines and Medical
Countermeasures
Naval Biomedical Reasearch Funding is all
soft money GRANT WRITING!!!
Know how to write a good grant
Most grant submissions will not be fundedlearn from your mistakes and resubmit
Funding cont.
• NIH funding lines- 7-10%
• DTRA funding lines- 4%
• Funding is becoming more
difficult and requires
tenacity
• Think outside the box for
potential funders
Funding- how are you going to be paid?
Post doc fundingNational Eye Institute, N.I.H., Individual National Research Service
EYO6844-02.
Award, P.I. "The role of the transcription factor CP2 in Allergic Eye Disease",
Junior faculty fundingNational Eye Institute, N.I.H., RO1 grant entitled "An Analysis of Antigen Presentation in a Mouse Model of Allergic Eye Disease" 1RO1EY12523-01.
NIH funding2000-2008. National Institute of Allergy and Infectious Diseases, N.I.H. Intramural funding.
BDRD/NMRC funding2008-2009 Defense Threat Reduction Agency (DTRA)/ Ichor. Electroporation for the Delivery of a Genetic Vaccine for Anthrax and Plague.
2008-2009 DTRA. Multivalent vaccine delivery platform based on Bacillus cereus spore display.
2008-2009 DTRA. Multi-epitope vaccines for anthrax, plague, melioidosis, Venezuelan equine encephalitis virus, and Ebola virus.
2008-2009 DTRA Development of a multiagent vaccine capable of conferring protection against anthrax, plague, botulinum neurotoxin and Marburg
virus
2008-2009. DTRA. Identification Of Conserved Genes Required For The Survival And/Or Virulence Of Intracellular Pathogens.
2009-2011 DTRA. Use of host miRNA signatures as a diagnostic indicator for the presence of B. anthracis, B. pseudomallei, and F. tularensis
infection.
2009-2011 DTRA. Development of a novel multi-agent vaccine against Bacillus anthracis, Yersinia pestis, and Ricin toxin using a Bacillus
thuringiensis Spore Display Platform
2009-2013 DTRA/ Syntiron Program. Multivalent Bacterial Vaccine Technology.
2010-2012 DARPA/ Ohio State University. Universal Vaccine Microparticulate Carrier that Encapsulates Immune Modifiers and Antigens in a Novel
polymeric Matrix to Passively Target Dendritic Cells
2012-2016 DTRA. Development of a needle –free, multi –formulation nanoparticle vaccine.
2013-2014 DTRA. Strategic Exploratory Development Award (SEED). A novel approach to deriving broadly protective vaccines against Burkholderia
pseudomallei challenge using Immune Refocusing Technology
What are my trainees currently doing?
(Traditional and alternative career paths)
• Academia•
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– Large universities (research with some teaching)
– Small colleges (primarily teaching)
FDA
NIH- grants managers
CIA
Medical Liaison
Private practice (M.D. or D.V.M.)
BSL-3/4 manager
Pharmaceutical Research
Patent Law
Military/ PHS
Work/life balance
Questions?