BiodefenseBiodefense / Human / Human Threats and Mass Threats and Mass

Spectrometry Spectrometry ApplicationsApplicationsGerman German HenostrozaHenostroza MDMD

Division of Infectious Diseases and Division of Infectious Diseases and International MedicineInternational Medicine

SARS(SevereSARS(Severe Acute Respiratory Acute Respiratory Syndrome)Syndrome)

ColeraColera

ColeraColeraGram negative bacteria causes Gram negative bacteria causes

severe watery diarrhea.severe watery diarrhea.Severe dehydration.Severe dehydration.Incubation period 24 hoursIncubation period 24 hoursJanuary 1991 to September January 1991 to September

1994 1994 -- Outbreak in Outbreak in South South AmericaAmerica, apparently initiated , apparently initiated when a ship discharged ballast when a ship discharged ballast water. Beginning in water. Beginning in PeruPeru there there were 1.04 million identified were 1.04 million identified cases and almost 10,000 deaths.cases and almost 10,000 deaths.

Plague : The Black DeathPlague : The Black Death

Ebola /MarburgEbola /Marburg

SmallpoxSmallpoxErradicatedErradicated ; 05/1980; 05/1980

Last case Birmingham , England after Last case Birmingham , England after which all isolates were sent to reference which all isolates were sent to reference labs for destruction.labs for destruction.

In 2003 Smallpox scabs found in Civil In 2003 Smallpox scabs found in Civil War Medicine book in Santa Fe, New War Medicine book in Santa Fe, New Mexico, no cases reported.Mexico, no cases reported.

? used as biological weapon in French ? used as biological weapon in French and Indian wars, American and Indian wars, American Revolutionary war and perhaps World Revolutionary war and perhaps World War II (research).War II (research).

Incubation period 10 days , if severe Incubation period 10 days , if severe disease death may happen in 3disease death may happen in 3--5 days.5 days.

Vaccine available but hypothesized that Vaccine available but hypothesized that virus might be genetically modified for virus might be genetically modified for use as use as bioweaponbioweapon..

DETECTION OF BIOATTACK OR OUTBREAK

RELEASE

PROGRESSION

SYNDROME Disease SURVEILLANCEBioSense ProMed

PATHOGEN in AIR PATHOGEN in PEOPLEBioWatch

Human:

Bug:

DETECTION OF BIOATTACK OR OUTBREAK

RELEASE

PROGRESSION

SYNDROME Disease SURVEILLANCEBioSense ProMed

PATHOGEN in AIR PATHOGEN in PEOPLE

PRE-SYMPTOMATIC HOST-BASED DETECTION

Advantages of HostAdvantages of Host--BasedBasedPrePre--Symptomatic DetectionSymptomatic Detection

Treat to PreventTreat to Prevent

Containment of OutbreakContainment of Outbreak

Triage Based on DiagnosisTriage Based on Diagnosis

Solution to AGENT X ProblemSolution to AGENT X Problem

DUAL USE TECHNOLOGYDUAL USE TECHNOLOGY

Biosignature Pattern Recognition in Human DiseasesHost-based Presymptomatic Detection of Events

Genes

Environment

Age

Sex

Serum/Cell components

State of Health

Center for Innovations in Medicine

Doc In Box: BioSignatures

prognosis

treatment strategy

diagnosis

PresymptomaticDiagnosis

clinical validation

drugs/vaccines

ILL

NOT ILLILL

NOT ILL

Signal

Signal

DiagnosticDevice

Center for Innovations in Medicine

Upper Respiratory DiseaseUpper Respiratory DiseaseIncubation PeriodsIncubation Periods

RhinovirusesRhinoviruses

InfluenzaInfluenza

ParainfluenzaParainfluenza

Respiratory Respiratory SyncytialSyncytial Virus (RSV)Virus (RSV)

PertussisPertussis

AdenovirusAdenovirus

EpsteinEpstein--Barr virus (EBV)Barr virus (EBV)

I I I I I I I I I I I I I I I I I I I I I I I I I I I 1 5 10 15 20 Days

4-6 Weeks

5-8 Days

7-10 Days

7 Days

4 Days

1-4 Days

5 Days

Sources: http://www.emedicine.com/med/topic2339.htmhttp://virus.stanford.edu/adeno/adeno.htmlhttp://www.cdc.gov/flu/professionals/diagnosis/

Goal:Goal:Characterize the pathogen and/or host cell proteome, Characterize the pathogen and/or host cell proteome,

identifying proteins associated with biology of identifying proteins associated with biology of microbes, mechanisms of microbial pathogenesis, microbes, mechanisms of microbial pathogenesis, and host response to infection.and host response to infection.

Discover targets for potential candidates for the next Discover targets for potential candidates for the next generation of vaccines, therapeutics, and generation of vaccines, therapeutics, and diagnostics.diagnostics.

Proteomic Technology DevelopmentProteomic Technology Development

Albert Einstein College of MedicineAlbert Einstein College of MedicinePI: PI: Ruth Ruth AngelettiAngelettiPathogens:Pathogens: ToxoplasmaToxoplasma gondiigondii, Cryptosporidium , Cryptosporidium

parvumparvum

Myriad Genetics, IncMyriad Genetics, IncPI: PI: Jerry Jerry LanchburyLanchburyPathogens:Pathogens: Bacillus Bacillus anthracisanthracis, , YersiniaYersinia pestispestis, ,

FrancisellaFrancisella tularensistularensis, , vacciniavaccinia, , variolavariola

The Scripps Research InstituteThe Scripps Research InstitutePI:PI: Peter KuhnPeter KuhnPathogens:Pathogens: SARSSARS--CoVCoV

University of MichiganUniversity of MichiganPI:PI: Phillip HannaPhillip HannaPathogen:Pathogen: Bacillus Bacillus anthracisanthracis

Harvard Medical SchoolHarvard Medical SchoolPI: PI: Joshua Joshua LaBaerLaBaerPathogens: Pathogens: Bacillus Bacillus anthracisanthracis, , VibrioVibrio choleraecholerae

CaprionCaprion Pharmaceuticals, Inc Pharmaceuticals, Inc PI: PI: Eustache Eustache ParamithiotisParamithiotisPathogen: Pathogen: BrucellaBrucella abortusabortus

Pacific Northwest National LaboratoryPacific Northwest National LaboratoryPI:PI: Richard SmithRichard SmithPathogens: Pathogens: OrthopoxOrthopox ((vacciniavaccinia and and

monkeypoxmonkeypox),), Salmonella Salmonella typhimuriumtyphimurium andandSalmonella Salmonella typhityphi

Administrative Resource CenterAdministrative Resource CenterPI:PI: Margaret MooreMargaret Moore

www.niaid.nih.gov/dmid/genomes/prc/default.htm

Overall SummaryOverall Summary--May 2006May 2006

Proteins Identified:Proteins Identified:

6 structures solved for SARS6 structures solved for SARS--CoVCoV; 282 human proteins that interact with ; 282 human proteins that interact with Y.Y.pestispestis(60), (60), B.B. anthracisanthracis(50), and vaccinia(172) proteins; 176 novel proteins (50), and vaccinia(172) proteins; 176 novel proteins identified from identified from B. B. abortusabortus, and 360 from , and 360 from T. T. gondiigondii..

Genes Identified:Genes Identified:

493 differentially expressed 493 differentially expressed murinemurine macrophage genes found upon infection macrophage genes found upon infection with anthraxwith anthrax

Reagents GeneratedReagents Generated

2983 2983 V. V. choleraecholerae Gateway entry clones made and deposited in an NIAID Gateway entry clones made and deposited in an NIAID repository for use by the communityrepository for use by the community

Progress AchievedProgress Achieved ––May 2006May 2006

Proteins IdentifiedProteins Identified

360 membrane and 360 membrane and cytoskeletalcytoskeletal proteins identified in proteins identified in T. T. gondiigondii

176 outer membrane proteins associated with virulence identified176 outer membrane proteins associated with virulence identified in in B. B. abortusabortus

282 human proteins found to interact with 282 human proteins found to interact with vacciniavaccinia (172), (172), B. B. anthracisanthracis (50), and (50), and Y.Y.pestispestis(60) proteins(60) proteins

6 SARS6 SARS--CoVCoV structures solvedstructures solved

2121/2343 proteins identified in virulent/2121/2343 proteins identified in virulent/avirulentavirulent S. S. typhimuriumtyphimurium

263 early response and 329 late response proteins identified in 263 early response and 329 late response proteins identified in murinemurine macrophages macrophages infected with anthraxinfected with anthrax

Speciation/IdentificationSpeciation/Identification

18% of GNP on Health Care in 2006 Projected to be 25% - 30% by 2015

Average annual healthcare Average annual healthcare expenditures by age, 2005expenditures by age, 2005

$0

$500

$1,000

$1,500

$2,000

$2,500

$3,000

$3,500

$4,000

$4,500

All ages Under25

25-34 35-44 45-54 55-64 65+ 65-74 75+

Age

Source: U.S. Bureau of Labor Statistics, U.S. Department of Labor. Consumer Expenditures in 2005. Report 998, Feb 2007.

Total Population vs. 65+ Population, 1950Total Population vs. 65+ Population, 1950--20502050

0

100,000,000

200,000,000

300,000,000

400,000,000

500,000,000

U.S

. Pop

ulat

ion

(n)

1950

1960

1970

1980

1990

2000

2004

2005

2010

2020

2030

2040

2050

All ages65 years and over

Source: U.S. Bureau of the Census, multiple Census years.

20.7% of the total population in 2050 is estimated to be over 65 years old.

*Only 8.1% of the total population in 1950 was over 65 years old.

What are the Costs of Medicine?What are the Costs of Medicine?

$2.26T/Year*Drugs

DiagnosticsCare

Source: Centers for Medicare & Medicaid Services, Office of the Actuary, 2007. Healthcare spending projections are based on the 2005 version of the National Healthcare Expenditure data released by the CMS in January 2007. Borger C, et al. “Health spending projections through 2015: changes on the horizon.” Health Affairs. March/April 2006; 25(2): 61-73.

*Estimated health expenditures for 2007

The Cost of PostThe Cost of Post--symptomatic Medicinesymptomatic Medicine

~$2 Trillion in Direct Medical Costs per Year

Patient Care88%

Drugs10%

Diagnostics2%

Two Options to Solve HC CrisisTwo Options to Solve HC Crisis

REDUCE CARE PROVIDEDREDUCE CARE PROVIDED

oror

TRANSITION TO PRETRANSITION TO PRE--SYMPTOMATIC SYMPTOMATIC DIAGNOSIS AND PREVENTATIVE DIAGNOSIS AND PREVENTATIVE MEDICINEMEDICINE

1960 1965 1970 1975 1980 1985 1990 1995 2000 2005 2010 2015 2020 2025 2030 2035 2040

Time

Risk

BioRevolution

Natural Pathogens

AntibioticResistance

DecisionDriven Δ

Valley of theShadow ofDeath

GenomeSequencing

DNA Shuffling

Chemical GeneSynthesis

Reverse Viral Genetics

RNAi

RecombinantDNA

Changing Spectrum of Changing Spectrum of BioThreatBioThreat RiskRisk

TuberculosisTuberculosis

TB TimelineTB Timeline

Exposure to Infectiouspatients

No infection (70%)

Infection (30%)

Early progression (5%)

Containment (95%)

Late progression (5%)

Continued containment (90%)

Nonimmunologicdefenses

Immunologicdefenses

Immunologicdefenses

Diagrammatic representation of the consequences of exposure to M. tuberculosis. The percentages shown are derived from epidemiologic data in persons without recognized immune response

•Three important steps:

•Phagosome formation

•Granuloma formation

•Failure of containment

TB Pathogenesis in a “Nutshell”

• Recruitment of cells induced by CC and CXC production.

•TNF-α dominant CC. Produced by infected macrophages and induced production of other CC and CXC.

• Subsequent recruitment of cells and production of IFN-γ

Cytokine / Chemokine Storm

Cell Mediated Immunity

TUBERCULOSIS TUBERCULOSIS AND PROTEOMICSAND PROTEOMICS

0.0 5.0k 10.0k 15.0k 20.0k

0

1000

2000

3000

4000

5000

6000

Nor

mal

ized

Rel

ativ

e In

tens

ities

m/z

Control TB TB/HIV

13.80k 13.85k 13.90k 13.95k

0

200

m/z

11.2k 11.4k 11.6k 11.8k 12.0k

0

m/z

4.92k 4.95k 4.97k 5.00k0

1000

m/z

11.2k 11.4k 11.6k 11.8k 12.0k

0

Nor

mal

ized

Rel

ativ

e In

tens

ities

m/z

Control TB TB/HIV

13.80k 13.85k 13.90k 13.95k

0

200

Nor

mal

ized

Rel

ativ

e In

tens

ities

m/z

Control TB TB/HIV

4.92k 4.95k 4.97k 5.00k0

1000

Nor

mal

ized

Rel

ativ

e In

tens

ities

m/z

Control TB TB/HIV

2007

3312

3953

4051

4151

4208

4465

4787

7764

8125

8125

8601

8931

928711

52511

68112

45013

879

0

5000

10000

15000

20000

25000

30000

35000

40000

45000ControlTBTB/HIV

m/z

Nor

mal

ized

Rel

ativ

e In

tens

ities

MDS

MDS - TB Vs. TB/ HIV

MDS - Control Vs. TB/ HIV

MDS - Control Vs. TB

Medicine: Art to ScienceMedicine: Art to ScienceSir William Osler 1892

“If it were not for the great variability among individuals, medicine might be a science, not an art”

Because of our new ability to measure variability among individuals, medicine now can become a science rather than an art.

Thank youThank you