dso - Harvard University Department of Physicsusers.physics.harvard.edu/~wilson/pmpmta/papers_for... · against Biological threats ... Ł Experimental/Theoretical Characterization

Dr. Mildred A. DonlonProgram Manager,Defense Advanced Research Projects AgencyDefense Sciences Office3701 North Fairfax DriveArlington, VA [email protected]

http://www.darpa.mil/dso

All slides have been cleared for public release

22

Director, Tony Tether

Defense SciencesMichael Goldblatt

Steven Wax

Bio Warfare Defense TechnologiesBiologyMaterials & DevicesMathematics

Information ProcessingRonald BrachmanZachary Lemnios

Architectures & DesignsComputer Processing & StorageNetworks Human Computing

Interfaces

MicrosystemsTechnologyRobert Leheny

John Zolper

ElectronicsOptoelectronicsMEMSCombined

Microsystems

Tactical TechnologyAllen Adler Art Morrish

Air/Space/Land PlatformsUnmanned SystemsSpace OperationsLaser SystemsFuture Combat SystemsPlanning / Logistics

Advanced TechnologyDavid HoneyLarry Stotts

Assured C3ISRMaritimeEarly Entry/Special Forces

Special ProjectsAmy AlvingJoe Guerci

Chem/Bio Def SystemsCounter Underground FacilitiesSpace Sensors/StructuresNavigation/Sensors/ Signal Processing

Information ExploitationStephen WelbyRobert Tenney

Sensors

Exploitation Systems

Command & Control

Information AwarenessJohn Poindexter

Robert Popp

Asymmetric Threat

Prediction

Behavior Modeling

DARPA OrganizationDARPA Organization

33

DARPA AccomplishmentsDARPA Accomplishments

Taurus LaunchVehicle

Global Hawk

MALD

Predator

MEMSTMR

Uncooled IR

BAT

JSF

19901990

UCAV

20002000

Vela HotelSaturn

M-16 Rifle

GroundSurveillance Radar

JSTARS

ATACMS

19601960

19701970

ArpanetStealth Fighter

Sea Shadow

19801980

GPS

44

Harvesting Biology for Revolutionary Technologies at DARPA

Harvesting Biology for Revolutionary Technologies at DARPA

EnhancedHuman Performance�Making People More Than They Can Be�

Biocomputation

New Materials

Genomics &

proteomics

The bio-silico interface

Brain Technology

Bioinformatics

Behavior

Energy Transduction

Infrastructure

Enhanced System Performance

�Using Biology to Enhance

Defense Systems�

Protecting Human Assets

�Defending against

Biological threats�

55

Biology is a major driver for new math and physics

Biology is a major driver for new math and physics

• Interrogate and manipulate biological systems with modern physical devices

• Analyze, model, simulate & design with a new arsenal of math and computational tools

10 nmout

in

+++ ++

+

77

88

NEAR MID FAR

Scie

nce

& T

echn

olog

y $

Service S&T

Fundamental Research

DARPA Role in Science and TechnologyDARPA Role in Science and Technology

99

NEAR MID FAR

Scie

nce

& T

echn

olog

y $

Service S&T

Fundamental Research

DARPA

DARPA Role in Science and TechnologyDARPA Role in Science and Technology

1010

NEAR MID FAR

DARPA OFFICES

Defense SciencesInformation ProcessingMicrosystems Technology

Advanced TechnologySpecial ProjectsTactical Technology

Information ExploitationInformation Awareness

Internal Technology FlowInternal Technology Flow

1111

BioWarfare ThreatBioWarfare Threat

Update: The Trend Lines

• The growing threat of BW is generated by three disturbing trends:� The number of players possessing biological

weapons or seeking to acquire a biological warfare capability is increasing.

� Biological agents with increasing lethality are being developed.

� Detection of BW programs and the acquisition of BW-related capabilities is difficult.

• It happened here

National Intelligence Council Report: The Biological Warfare Threat, Jan 2001.

1212

Protecting Human AssetsProtecting Human Assets

Genome Sequencing

Sensors Protection Collective/

Building

Consequence Management

Advanced Diagnostics

Medical Countermeasures

Decontamination

Years Seconds to minutes

Minutes to hours

Minutes to hours

Hours to days

Hoursto days

Daysto years

1313

Simbiosys ProgramAnantha Krishnan, DSO

Novel Tools (Devices, Models, Databases, Algorithms ) for High Performance Biological/Chemical Sensing and

Processing

+

Simbiosys

Novel Processes, Experimental Methods, Computational Models and Design Tools that

significantly improve speed, sensitivity, specificity and efficiency of chemical/biological

processing and analysis

Novel Processes, Experimental Methods, Computational Models and Design Tools that

significantly improve speed, sensitivity, specificity and efficiency of chemical/biological

processing and analysis

Challenges� Experimental/Theoretical Characterization of the elements of a Bio-Molecular System :

�Molecular Recognition Elements (sensitivity, selectivity and speed)�Signal Transduction Elements (Signal Amplicationwith High SNR)�Bio-Fluidic Transport Elements (High Efficiency, Low Power Pumping, Valvingand Mixing Methods)

� Demonstration of device modelson design of Bio-Microsystems

Interface Between Biology and Micro/Nano-Technology

1414

p - AlGaNAlInGaN MQW

n - AlGaN

GaN buffer layer

Sapphire substrate

n - electrode

Grading Layer

n+ - GaNp+ - GaN

n - electrode

SUVOS - Semiconductor Ultraviolet Optical Sources John Carrano, Program Manager, MTO

Develop Solid-State UV Sources down to 280 nmCompared to conventional technology• Power consumption reduced by 50X• Size & weight reduced by 100X

LEDs andLaser Diodes:The Next Step

in efficientUV powergeneration

Enabling new classes of bio-agent detection systems

Emitter -Detector

sensor arrays

Bio-agentfluorescent signatures

Bio-Alertin every office

Visible LightUltraviolet

high energy Wavelength (nm) low energy200 300 400 500 600 700 800 900

Aerosol samplers �small and efficientfor MAV deployment

Low-co

st

mass p

roduc

ible

SUVO

S so

urce

s

100X

sm

aller

50X

mor

e ef

ficien

t

UV LED

Infrared

1515

M O S A I CMolecular Observation, Spectroscopy And Imaging using Cantilevers

Atomic Force Microscopyatomic scale, non-destructive,control

but only surface

Magnetic Resonance Imaging3D, non-destructive

but macroscale

Magnetic Resonance Force Microscopy3D, atomic scale, non-destructive, and with control

Technical hurdles� Signal processing� Nanofabrication� Readout

1616

BIOSBIOS

Techbase program to make plug & play (PnP) biocircuits that enable engineering of organisms for use as sentinel devices

1717

Bacteria enterthe cell in body (lung) or sensor

Bacteria invade alveolar cells

Bacteria internalize and multiply

Macrophage

Activity Based Detection: A Conceptual Framework

Alan Rudolph, DSO

Cellular or Tissue activity can be used as diagnostic or to engineer markers for sensors

Evaluate Activated Pathways

for Sensor

Report Infection

for Medical

Diagnostic

In vitro

In vivo

1818

Signs/Symptoms

100

Joint Pain

Cough

Chills

Vomiting

AppetiteLoss

Fever

Headache

Frequency (%)0 10 20 30 40 50 60 70 80 90

{{{{{{{{

Muscle Pain

TularemiaTreatment: Antibiotics -May be fatal if untreated.

Advanced Diagnostics�Why We Need It

Advanced Diagnostics�Why We Need It

No treatment currently available - Incapacitating (can cause severe disease at high doses).

SEB Toxicity

Treatment: Antibiotics -Incapacitating, not usually fatal.

Q-Fever

Early symptoms of many BW agents are flu-like and indistinguishable

1919

Signatures of ExposureSignatures of Exposure

Program Goal: Determine cell signatures of early exposure to BW pathogens and develop rapid signature diagnostics

Concept: Generate differential expression profiles and cDNA libraries from cells after exposure. Identify differentially expressed transcripts. Create gene reporter elements for diagnostic tests

WRAIR,Georgetown University, Stanford, Umea Sweden, APL

0

50

100

150

200

250

300

350

1 2 3 4

SEBAnthrax

0

100

200

300

400

500

600

700

800

900

1 2 3 4

SEBAnthrax

Changes in Expression of a GBP (unique to SEB vs. other toxins tested)

Changes in Expression of HIF-1 (common to many toxic agents)

SEB

Anthrax

SEB

Anthrax

2020

Unconventional Pathogen CountermeasuresJohn Carney, DSO

Unconventional Pathogen CountermeasuresJohn Carney, DSO

Technical Challenges:! Develop therapeutics effective

against all known and unknown (e.g. engineered) BW pathogens

! Develop effective multivalent vaccines with broad efficacy versus industry standard one bug one drug

! Enhance Immune system in order to eliminate the need for vaccines

Idea: Develop broad spectrum technologies that effectively counter potential BW threats to the US military and civilians

BroadBroad--Spectrum ApproachesSpectrum Approaches

Inhibition of Pathogen GrowthMIC (µM)

12.5MW6-15a

12.5SB28b

12.5SB20e

12.5DW522b

12.5DW523

6.25MW5304

6.25SB28d

106.25SB28c

6.25SB220b

6.25106.25DW520b

256.25DW522

1006.25MWTP

12.56.25MW6-15

12.53.125DW520

3.125103.12528C

Helicobacter pyloriBrucella abortusBacillus anthracisCompound

kills intracellular Brucella

not detrimental to cultured macrophages at >100µm

AntibacterialAntibacterial

Ibis Drug Discovery Strategy for RNA

Gene sequence data

RNA Subdomain Selection

Animal Studies

Screening

MASS Assay System HTS Functional assays

Lead Compound

DARPA Task 1

DARPA Task 2

DARPA Task 3,4

DARPA Task 5

USAMRIID Transition

AntiviralAntiviral

AntiAnti--Genomic Drug Binds to AGenomic Drug Binds to A--T rich T rich Targets and Blocks Essential Pathogen Targets and Blocks Essential Pathogen

GenesGenes

GeneSoft, Inc.

Essential Essential GeneGene




AnthraxAnthraxTularemiaTularemia

Small PoxSmall Pox

PneumoniaPneumonia

MalariaMalaria


AntigenomicsAntigenomics

Plants for Production of Vaccines and Antibodies

Edible plant vaccines stimulate immune responsesNature Medicine 4: 607-609 (1998)

Plant-expressed Abs provide protection in humansNature Medicine 4: 601-606 (1998)

Plants are an economical production system for subunit vaccine proteins to prevent infectious disease, and for antibodies to treat diseases or toxin exposure.

Bacterial or ViralStructural Gene

VaccinesVaccines

Breeding of Cross-Reactive Antigens

Natural evolution

Crea te reactivity aga ins t severa l antigenic variants

Exis ting Varia nts

Combine e pitopes

Ances tra l Molecule

VaccinesVaccines

CpG DNACpG DNA

MPMPDCDC

AdaptiveAdaptive protective immune protective immune responses: antigenresponses: antigen--specific antibodies (B cells), specific antibodies (B cells), killer T cells (killer T cells (slow, slow, enhance vaccinesenhance vaccines))

BBTT

NKNK InnateInnate protective immune protective immune responses: cytokines, responses: cytokines, chemokines chemokines ((immediate protectionimmediate protection))

Kill infected cellsKill infected cells

CpG DNA Activates Broad Spectrum Immune DefensesCpG DNA Activates Broad CpG DNA Activates Broad

Spectrum Immune DefensesSpectrum Immune Defenses

IFNIFN--γγ

CpG DNA is detected by immune cells CpG DNA is detected by immune cells (macrophage [MP], dendritic cell [DC], (macrophage [MP], dendritic cell [DC],

natural killer [NK])natural killer [NK])

Once alerted to �danger� by Once alerted to �danger� by injection of CpG DNA, the injection of CpG DNA, the activated immune system activated immune system protects the subject from protects the subject from infections for at least 2 weeksinfections for at least 2 weeks

ImmunotherapyImmunotherapy

PathogenesisPathogenesisRole of Host in Susceptibility to Viral Pathogens

(Why do I not catch foot and mouth disease from my cattle?)

Disease

binding

genome internalization

gene expression

protein processing

viral replication

pathogenic event

morphogenesis & release

Virus

Host

BroadBroad--Spectrum ApproachesSpectrum Approaches

Inhibition of Pathogen GrowthMIC (µM)

12.5MW6-15a

12.5SB28b

12.5SB20e

12.5DW522b

12.5DW523

6.25MW5304

6.25SB28d

106.25SB28c

6.25SB220b

6.25106.25DW520b

256.25DW522

1006.25MWTP

12.56.25MW6-15

12.53.125DW520

3.125103.12528C




AntibacterialAntibacterialInhibition of Pathogen Growth

MIC (µM)

12.5MW6-15a

12.5SB28b

12.5SB20e

12.5DW522b

12.5DW523

6.25MW5304

6.25SB28d

106.25SB28c

6.25SB220b

6.25106.25DW520b

256.25DW522

1006.25MWTP

12.56.25MW6-15

12.53.125DW520

3.125103.12528C




AntibacterialAntibacterial

Ibis Drug Discovery Strategy for RNA

Gene sequence data


Animal Studies

Screening


Lead Compound

DARPA Task 1

DARPA Task 2

DARPA Task 3,4

DARPA Task 5

USAMRIID Transition

AntiviralAntiviralIbis Drug Discovery Strategy for RNA

Gene sequence data


Animal Studies

Screening


Lead Compound

DARPA Task 1

DARPA Task 2

DARPA Task 3,4

DARPA Task 5

USAMRIID Transition

AntiviralAntiviral

AntiAnti--Genomic Drug Binds to AGenomic Drug Binds to A--T rich T rich Targets and Blocks Essential Pathogen Targets and Blocks Essential Pathogen

GenesGenes

GeneSoft, Inc.






Small PoxSmall Pox

PneumoniaPneumonia

MalariaMalaria


AntigenomicsAntigenomicsAntiAnti--Genomic Drug Binds to AGenomic Drug Binds to A--T rich T rich

Targets and Blocks Essential Pathogen Targets and Blocks Essential Pathogen GenesGenes

GeneSoft, Inc.






Small PoxSmall Pox

PneumoniaPneumonia

MalariaMalaria


AntigenomicsAntigenomics

Plants for Production of Vaccines and Antibodies





VaccinesVaccinesPlants for Production of Vaccines and Antibodies





VaccinesVaccines

Breeding of Cross-Reactive Antigens

Natural evolution


Exis ting Varia nts

Combine e pitopes


VaccinesVaccinesBreeding of Cross-Reactive Antigens

Natural evolution


Exis ting Varia nts

Combine e pitopes


VaccinesVaccines

CpG DNACpG DNA

MPMPDCDC


BBTT





IFNIFN--γγ





CpG DNACpG DNA

MPMPDCDC


BBTT





IFNIFN--γγ







Disease

binding


gene expression

protein processing

viral replication

pathogenic event


Virus

Host



Disease

binding


gene expression

protein processing

viral replication

pathogenic event


Virus

Host

Highlights:" 5th year of funding" USAMRIID technology transition partner

(10 transitions to date, 6 under discussion)" Possible development of 5 new

pharmacological therapeutics

2121

MicroVIC® Bio-aerosol Concentrators MicroVIC® Bio-aerosol Concentrators

• Technology:� Virtual Impaction (aerodynamic

sorting and concentrating)� Developer: MesoSystems

• Key Features:� Low power� Compact and versatile

� Wet or dry particle collection� Wet collection� Aerodynamic pre-filtering� Particle pre-concentrator for

triggers� Mass-producible (low cost)

• Commercialization in progress

Bioni� UV trigger byPacific Scientific Instruments

Continuous Air Samplerfor Lockheed Martin

0%

20%

40%

60%

80%

100%

0 2 4 6Particle Diameter (microns)

Sepa

ratio

n Ef

ficie

ncy

Experimental Computational

2222

BioBadge Wearable Bio-dosimetryBioBadge Wearable Bio-dosimetry

• Technology:� MesoSystems Rotating Impactor

combined with SRI International�s UPT Immunoassay

� Field exposure quantification for the individual

• Key Features:� Low power; field operation� Compact and light� 15-30 liters/min flow rate� 40% overall efficiency from Air

# Disk # Liquid sample• Commercialization status

� Laboratory testing in 2002� Field trials in 2003

0%

20%

40%

60%

80%

100%

Biological SamplerEfficiency

Extraction StationEfficiency

System Efficiency

none coating 1 coating 2 coating 3Coating:

2323

Personal Water Purification SystemsPersonal Water Purification Systems

Cryptosporidium parvum Inactivation with MIOX Disinfection Pen

-8-7-6-5-4-3-2-1012

0 10 20 30 40 50 60 70 80 90 100Time in minutes

Log

(Nt/N

o)

Beta Pen

Alpha Pen

Bleach

US EPA Purifier Standard

� New Generation Hydration System� Lightweight/reduced footprint for �on-the-move� operation (backpack)� Produces Gatorade-like nutrient drink from contaminated water via FO bag� MIOX pen/cap inactivates even extremely resistant microorganisms by 99.99%

� Tech Approach:� Effective mixed-oxidant chemistry from table salt� Smart electrochemical cell and circuit � Miniaturization of mechanical filters and purifiers� Pen/pump combo treats any water source� Low cost to produce� Working turbidity filtration issues

MIOX Disinfection Pen/Cap

Reverse Osmosis (RO)Pump / Advanced Membranes

2424

• FY03: Site selection

• FY04: System design and testing; on-site characterization

• FY05: Demonstration

Implement, optimize, and demonstrate a full-scale building protection system at a US military site

Immune Building Demonstration

Immune Building Demonstration

2525

Integrated Systems Experimentation Team

Integrated Systems Experimentation Team

Fort McClellan, Anniston, AL

• Battelle

• Auburn University

• Ensco, Inc.

• Fidelity Engineering Corp.

Contact: James E. Risser, [email protected]: James E. Risser, [email protected]

� Gage-Babcock& Associates

2626

Bio Sensors ContextBio Sensors Context

Antibodies

hours minutes seconds

Surveillance

Det

ectio

n ab

ility agent specific

reducedfalse alarms

bio particles

any particles

threat particles

MassSpec

weeksCapabilitytoday

CulturePCR

APS

?

Goal

Forensics Early warning

2727

Biosensor Technologies

Upconverting Phosphor Technology(UPT)-SRI International (Menlo Park, CA)Handheld Biosensor

-University of Washington -Argonne National Laboratory (ANL)-JHU/APL - Food and Drug Administration (FDA)

(DARE) DARPA Antibody ReplacementEnhancement Team

-University of Alabama-Archemix-University of Texas at Austin-Polytechnic University-University of Missouri-Auburn University

Ion-channel Biosensor Team-Auburn University -Sandia National Laboratory-Rush-Presbyterian -Polytechnic University

Micro Array of Gel-Immobilized CompoundsMAGIChipTM

IdentifyingTechnology

Critical Sensor Technology

ANTIBODY

Documents

dso - Harvard University Department of Physicsusers.physics.harvard.edu/~wilson/pmpmta/papers_for... · against Biological threats ... Ł Experimental/Theoretical Characterization