Jane E. Gallagher Ph.D. - EPA ArchivesJane E. Gallagher Ph.D. Human Studies Division ORD NHEERL EPA September 24-25 2007 Public Health Applications of Human Biomonitoring Mechanistic

Jane E. Gallagher Ph.D.Jane E. Gallagher Ph.D.Human Studies DivisionHuman Studies DivisionORD NHEERL EPAORD NHEERL EPASeptember 24September 24--25 2007 25 2007 Public Health Applications of Human Public Health Applications of Human BiomonitoringBiomonitoring

Mechanistic Indicators of Childhood Asthma (MICA) -Integrating Environmental, Clinical and Susceptibility Markers to Improve the Impact of Human Air Pollution Studies.

Advances in biomarker development haveimproved our ability to detect early changes at the molecular and cellular level.

C lin ic a lD is e a s e

E x p o s u re

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Battery of endpoints Battery of endpoints capturing Net effectcapturing Net effect––over over several mechanisms of several mechanisms of

action/ classesaction/ classesComet assayComet assayP53 P53 FISHFISH11--OH OH pyrenepyreneCholinesterase Cholinesterase inhibition inhibition CrossCross--linking metals linking metals formaldehyde formaldehyde

HPRTHPRTDNA adducts DNA adducts methodsmethodsMutagen sensitivity Mutagen sensitivity Oxidative damage Oxidative damage MutagenicityMutagenicity

Integrated measure of dose across classes of chemicals

Environment Health Scientists

Clinicians

EvolvingTechnologies

C l i n i c a lD i s e a s e

E x p o s u r e

?

? ?

•Identify Genetic variants that increase susceptibility

Environmental Studies Benefit from Knowledge Gained from Clinical Disease Studies (and visa versa)

•What factors affect a persons risk for a number of health conditions.

•Determine whether the effect of genetic variants that increase risk is different in the presence of

environmental exposures

•Early indicators/detection of disease


E x p o s u re

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Biological Fluids/cells

MICA

Change in research paradigm

Work across disciplines Give up data for the greater

good communicate

NHEERL

Westat

Henry Ford HealthSystem

South West Research Institut

RTI

Johns HopkinsMichigan State University nMercy College UNC at Chapel Hill

Rutgers University Harvard U.

NERL RTP

NERLCincinnati.

ECD ETDHSD

Genomics Core

National Center for Computational Toxicology

Region 5

Contracts

S. Edwards

MICA-Change Research Paradigm

Labcorp

EAWestat

MICA MICA A childhood asthma study and Parallel A childhood asthma study and Parallel rodent studyrodent study

A NHEERL and NCCT Computational A NHEERL and NCCT Computational Toxicology study Toxicology study

Combines and Integrates biomarkers of Combines and Integrates biomarkers of exposure effects and susceptibility in the exposure effects and susceptibility in the context of clinical measurements and context of clinical measurements and disease (asthma) outcome. disease (asthma) outcome.


GoalsImprove linkages in the source to outcome paradigm

•Provide predictive models for Hazard ID

•Improve Quantitative Risk assessment Dose, species, chemical class

7 New Starts ---MICA

SOURCE HEALTH OUTCOME

? ??

INTERNALDOSE

INTEGRATEDDOSE

EARLYRESPONSES

RNA--- Blood Gene expression DNA ----11 genes 55 SNP

Asthma

urine bloodFingernails

Vacuum dust and

Passive monitoring

BMI Blood PressureO2 SaturationCell differentialsBlood ChemistryCytokinesCreatininecotinineMedications

Lung functionNO ex

QuestionnaireDietTime activity

Odor test

MICA

MICAMICA

Increase our understanding of asthma by assessing the complex gene/environmental relationships through the combined use of innovative methods to manage and analyze multifactorial data

Objective

11 polymorphic genes

55 SNP

MICA RODENT Phase 1 Childhood asthma Phase 2

MICA Nested Within Detroit MICA Nested Within Detroit ChildrenChildren’’s Health Study s Health Study

Questionnaire

Lung FunctionLung Function

BiomarkersBiomarkers

MICAMICA AIR

Detroit Exposure Aerosol Research Study “DEARS”

National Exposure and Research Laboratory

MICAMICA200 children ( asthma and no asthma) 9200 children ( asthma and no asthma) 9--1212

years of age years of age 100 families participated in self monitoring100 families participated in self monitoring(indoor outdoor) as part of MICA air (indoor outdoor) as part of MICA air Vacuum Dust and medication list brought to Vacuum Dust and medication list brought to clinicclinicEducational and Educational and ““station walk throughstation walk through””presentation to provide context to the studypresentation to provide context to the studyConsent assent and Questionnaire Consent assent and Questionnaire Lung function, NO ex and odor testingLung function, NO ex and odor testingBlood, urine, fingernails collected Blood, urine, fingernails collected

> 90 percent of subjects provided samples at > 90 percent of subjects provided samples at each station.each station.

MICA Childhood Study MICA Childhood Study Multiple Risk FactorsMultiple Risk Factors

Asthma

3

Obesity

CardiovascularRisk

1

2

Asthma studies by race

MICA 85% African American

Study Design

Biomarkers of Integrated Dose:Metals (Lead, Mercury),

Metabolites of Polycyclic Aromatic Hydrocarbons, Cotinine, and Creatinine

Biomarkers of Early Effect:Autoantibodies, Mutagenicity

Gene Expression Inflammatory Markers

Clinically-relevant Outcomes:

Allergies, Asthma, Respiratory Symptoms,

Lung FunctionNOex VOC

Asthmatic and nonasthmatic Children n=200

Rodents: Blood and Lung Tissue

Gene ExpressionProfiles

InflammatoryMarkers

Susceptibility Factors: Genetic Variation-RNA, DNA

AmbientLevels

Internal Dose

EffectiveDose

Early Biological

Effect

ClinicalDisease

Pre-ClinicalEffects

Particulate matter (PM)

(concentrated to 200-600µg/m3)

Ambient Levelsof PM and air

toxics

Detroit-areaUrban Air

Rodent Study Detroit AIRConcentrated

Air Particles Exposures

MSU Mobile Air Research Laboratory (AirCARE 1)

J Harkema

Presenter

Presentation Notes

Harvard-like concentrator

Air Particle Concentratorand Inhalation Exposure System for Laboratory Rodents

Animals are exposed to concentrated fine and ultrafine particles in specially designed shoebox cages

MICA (I) MICA (I) Evaluate Utility of Rodent models for Evaluate Utility of Rodent models for analyzing gene expression data in analyzing gene expression data in

childhood asthma study childhood asthma study

Brown Norway Rats

Air/Saline Air/Ova CAPS/Saline

CAPS/Ova

MSU

Study Design

Biomarkers of Integrated Dose:Metals (Lead, Mercury),

Metabolites of Polycyclic Aromatic Hydrocarbons, Cotinine, and Creatinine

Biomarkers of Early Effect:Autoantibodies, Mutagenicity

Gene Expression Inflammatory Markers

Clinically-relevant Outcomes:

Allergies, Asthma, Respiratory Symptoms,

Lung FunctionNOex VOC

Asthmatic and nonasthmatic Children n=200

Rodents: Blood and Lung Tissue

Gene ExpressionProfiles

InflammatoryMarkers

Susceptibility Factors: Genetic Variation-RNA, DNA

AmbientLevels

Internal Dose

EffectiveDose

Early Biological

Effect

ClinicalDisease

Pre-ClinicalEffects

Particulate matter (PM)

(concentrated to 200-600µg/m3)

Ambient Levelsof PM and air

toxics

Detroit-areaUrban Air

MICA

INTERNAL DOSE

INTEGRATED DOSE

Health EffectAsthma

AIR/DUST

Lung functionNOexAntioxidantsAllergen skin testingPlasma ROSCytokines Blood panel Cell surface markers

ROSCholinesteraseMutagenicity1 OH Pyrene Antibodies to nervous system Cotinineproteins Coagulation factors

Metals Heavy MetalsPAH metabolitesETS Pb HgNapthols Phenanthrols

Gene expression - RNA Genotyping –DNA ( 11 genes 55 SNP )

Indoor outdoorPAH VOC No2O3 AllergensMolds

Passive monitoring: N02, PAHs VOC, (indoor and outdoors)Vacuumdust

Metals, PAHs,aero-allergens mold, endotoxin)

Biomarkers--- Clinical and EnvironmentalUrine Cotinine Creatinine

Urine Metals: Mercury, Cadmiun, Arsenic, Chromium, Manganese, Nickel

Urine 1- hydroxpyrene (1-OH pyrene) Napthols, Phenanthrols, Hydrocarbon Metabolites phthalates

Urine Mutagenicity Assays

SerumAutoantibodies for nervous system proteins, Blood Chemistry, Total IgE and specific antibodies to common aero-allergens (multiscreen inhalant and food antibody series)- *dust mite, Cockroach , Mouse, Rat Urine Protein,

Plasma Reactive Oxygen Species, cytokines (IL4, 6 IL13), tumor necrosis factor- alpha, c-reactive protein, fibrinogen

WholeBlood

hematology panel, lead and mercury, Gene expression (RNA), glycosolated hemoglobin

Nails Mercury, Cadmium, Arsenic, Chromium, Manganese, Nickel

Serum IgE-inducing proteins associated with fungal exposures

11 polymorphic genes

55 SNP

X D a ta0 1 2 3 4 5 6 7 8 9 1 0 1 1 1 2 1 3 1 4 1 5 1 6

Y D

ata

0

5 0

1 0 0

1 5 0

2 0 0

X Data0 1 2 3 4 5 6 7 8 9 10 11

Y D

ata

0

50

100

150

200

250

300

% A1C vs diastolic p <.05

4.0

4.5

5.0

5.5

6.0

6.5

7.0

7.5

0 20 40 60 80 100 120 140

X D a t a0 1 2 3 4 5 6 7 8 9 1 0 1 1

Y D

ata

0

5 0

1 0 0

1 5 0

2 0 0

2 5 0

3 0 0

X Data

0 1 2 3 4 5

Y D

ata

-10

-5

0

5

10

15

20

25

Plot 1

s y s t o l ic D ia s to l ic b y g e n d e r ( f v s m ) a n d a g e

X D a t a

0 2 4 6 8 1 0 1 2 1 4 1 6 1 8 2 0 2 2

Y D

ata

0

2 0

4 0

6 0

8 0

1 0 0

1 2 0

1 4 0

1 6 0

X D a ta0 1 2 3 4 5 6 7 8 9 1 0 1 1 1 2 1 3 1 4 1 5 1 6

Y D

ata

0

5 0

1 0 0

1 5 0

2 0 0

relationship between % neut vs % eosin

R2 = 0.1321

0

20

40

60

80

100

120

0 5 10 15

eos % vs neut y

Linear (eos % vs neut y )

X Data

0 1 2 3

Y D

ata

0

2

4

6

8

10

12

14asthma med types taken on daily basis

Singular, 16%

albuterol , 42%

Flovent , 3%

advair, 29%

pulmicort , 10%

050

010

0015

0020

00To

tal S

erum

IgE

Low Moderate High Asthma severity

020

4060

8010

0ph

adia

top


010

2030

fx5e

food

scre

en


020

4060

80de

rmpt

erdu

stm

ite


020

4060

8010

0de

rmfa

rindu

stm

ite


010

2030

4050

catd

ande

repi

thel


010

2030

40do

gdan

der


02

46

8ge

rman

cock

roac

h


0.5

11.

52

mou

seur

inep

rot


05

1015

2025

ratu

rinep

rot


010

2030

aspe

rgill

fum

igat

us


05

1015

20cla

dosp

orhe

rbar

um


020

4060

80al

tern

aria

alte

rnat

a


05

1015

20pe

nici

llnot

atum


Male (Blue); Female (Green)

BMI vs systolic

80

90

100

110

120

130

140

150

160

10 20 30 40 50

% A1C vs diastolic p <.05

4.0

4.5

5.0

5.5

6.0

6.5

7.0

7.5

0 20 40 60 80 100 120 140

X Data

0 1 2 3

Y D

ata

0

2

4

6

8

10

12

14

NOW WHAT ???

CINCINNATI CHILDRENS HOSPITALStable

AsthmaticsUnStable

AsthmaticsNNORMAL

NASALEPITHELIAL

CELLS

HEAT MAP MICA

Biomarkers Exposure/Clinical indicators

Sub

ject

sDavid ReifNationalCenter for ComputationalToxicology

Elaine Hubal

Biomarker NeedsBiomarker NeedsExposure biomarkers in the context of clinical Exposure biomarkers in the context of clinical health indicators health indicators Mechanistic information test biological Mechanistic information test biological plausibility in rodents plausibility in rodents Validation of surrogate cells with target tissue Validation of surrogate cells with target tissue responsesresponses

Archiving of biological and environmental samples and measurements as new

technologies advance


E x p o s u re

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• High-data content technologies, elucidating the genetic and environmental basis for

toxicity and disease

Summary

Gene expression arrays

genes, pathways, and networks

computational

bioinformatic

statisticalanalysis.

Integration of Diverse Set of exposure, effects and susceptibility

AcknowledgementsAcknowledgements


Stephen EdwardsElaine Hubal

David Reif

National Health Exposureand Effects Laboratory

PHASE II MICA

Westat

Henry Ford Health System Clinic Henry Ford Health System-lab


MICA-HSD Ed HudgensGina Andrews Brooke HeidenfelderJeff InmonMary JohnsonDanelle LobdellPauline MendolaJim PrahScott RhoneyElizabeth SamsDCHS Lucas NeasAnn WilliamsNCCTElaine HubalDavid ReifAdministrativeWalter BreyerEdward StrubbleMike Ray QADebra WalshKay Williams

NERLShaibal MukergeeHaluk OzkaynakRon Williams Dan ValleroNERL CincinnatiStephen Vesper

Mercy CollegeH. El FawalJohns HopkinsRobert HamiltonJohn WisemanCarol SchultzHarvard UniversityRutgers UniversityTina FainUNC Stephen RappaportSuryamya W.SWRI David CaamanRTIFrank Weber Peter GrosheWestatKurt PatriziHenry Ford Health SystemClinical and Lab

Contracts Robin Harris Jennifer HillLenora Hilliard Student ContractorsChris GarlingtonChrissy LinSharon MyersPeter Stone

Genomics core Susan Hester Chris CortonNHEERL Office of the Director Stephen Edwards ECD David DeMariniRegion 5George BollwegJackie Nwai

IRB Richard HermanMonica Nees

ETD Gary HatchMarsha Ward et alKay CrissmanMcGee

Nurses HSDMary Ann .BassettDeb LevinTracy. MontillaWestatAndrea Ware

Michigan State U.J. HarkemaLori Bramble


Documents

Jane E. Gallagher Ph.D. - EPA ArchivesJane E. Gallagher Ph.D. Human Studies Division ORD NHEERL EPA September 24-25 2007 Public Health Applications of Human Biomonitoring Mechanistic