Anthony Macherone, Ph.D.Agilent TechnologiesThe Johns Hopkins University School of Medicine

Genetics Exposure

Wild CP. Cancer Epidem Biomarkers Prev, 2005;14:1847-1850Wild CP. Int J Epidem 2012;41:24–32

Christopher P. Wild (2005) defined the exposome

Rappaport SM, J Expo Sci Environ Epidemiol, 2011;21

The Internal Exposome Metabolism, Inflammation, Xenobiotics, Preexisting disease, Oxidative stress, Gut microflora

The External ExposomeIonizing radiation, food, air and water,

pollution, diet, drugs, stress, infections, behavior and lifestyle

Exposomics is the application of “omics” tools to study the exposome◦ The heart of exposomics is epigenetics◦ The goals are: Identify and differentiate geomarkers and biomarkers Determine disease causality Develop better predictive and preventative measures in

healthcare – personalized medicine

Rappaport, SM. J Expos Sci Env Epidemiol, 2011;21:5–9*Vineis P & Perera F. Cancer Epidemiol Biomarkers Prev, 2007;16:1954–1965

Meet in the middle

Identify the overlap between exposure markers and predictive biomarkers of disease:

Causal link of exposure and disease

Environmental Analysis

Data or knowledge driven

Targeted, non-targeted or hybrid multi-class, multi-

residue

Measure the exposome

EWAS

LC, GC and other technologies

Correlate environmental contaminate risk potential

with exposome data

Identify relevant disease biomarkers

Align, annotate, refine

Submit to the Wikisome

GIS mapping

Prioritization

to account for variation, multiple sampling (natural and biological) and technical replicates are needed

1 3 5 7 9 11 13 15 17 19 21 23 25 27 29 31 33 35 37 39

TCDD

1 3 5 7 9 11 13 15 17 19 21 23 25 27 29 31 33 35 37 39

PCB 81

1 3 5 7 9 11 13 15 17 19 21 23 25 27 29 31 33 35 37 39

TCDF

Property Value TCDD TCDF PCBRotatable bonds >=1 0 0 1Rotatable bonds >=2 0 0 0Rotatable bonds >=3 0 0 0Rotatable bonds >=4 0 0 0Rotatable bonds >=5 0 0 0Molecular weight >100 1 1 1Molecular weight >200 1 1 1Molecular weight >300 1 1 1Molecular weight >400 0 0 0Molecular weight >500 0 0 0Molecular weight >600 0 0 0LogP >-1 1 1 1LogP >0 1 1 1LogP >1 1 1 1LogP >2 1 1 1LogP >3 1 1 1LogP >4 1 1 1LogP >5 1 1 1LogP >6 0 0 1Polar surface area >50 0 0 0Polar surface area >60 0 0 0Polar surface area >70 0 0 0Polar surface area >80 0 0 0Polar surface area >90 0 0 0Polar surface area >100 0 0 0H-bond acceptors >=1 1 1 0H-bond acceptors >=2 1 0 0H-bond acceptors >=3 0 0 0H-bond acceptors >=4 0 0 0H-bond acceptors >=5 0 0 0H-bond acceptors >=6 0 0 0H-bond acceptors >=7 0 0 0H-bond acceptors >=8 0 0 0H-bond acceptors >=9 0 0 0H-bond acceptors >=10 0 0 0H-bond doners >=1 0 0 0H-bond doners >=2 0 0 0H-bond doners >=3 0 0 0H-bond doners >=4 0 0 0H-bond doners >=5 0 0 0

Exposome-wide association studies◦ Discover components of the exposome that cause

complex chronic diseases◦ Establish a molecular basis for environmental

causes of disease Carefully conducted EWAS will:◦ Differentiate biomarkers of exposure from

biomarkers of disease◦ Determine disease causality◦ Offer diagnostic reference points◦ Guide treatment and patient care

Coordinated effort to define and develop exposome research◦ Currently building a consortium, “think tank” Define exposomics roadmap

Public repository for exposome data / information◦ Controlled, regulated and reviewed◦ Multi-disciplinary, multi-technique, open access

GIS mapping of environmental, exposome and public health data◦ Spiders to pull data into a single source Digimine, radian6

Environmental screening and

annotation

Exposome measurements

Identify geomarkers and biomarkers:

Causal and Reactive links

Fluorotelomer alcohols induce hepatic vitellogenin through activation of the estrogen receptor in male medaka (Oryzias latipes)◦ Vitellogenin: biomarker for estrogen

EDC exposure◦ Dose-dependent interaction between

medaka estrogen receptor alpha (ERalpha) and coactivator TIF2

Ishibashi H, Yamauchi R, Matsuoka M, et al. Chemosphere 2008;71(10):1853-9.

L. Levi, I. Pekarski, et al. BMC genomics, Retrieved May 10, 2013 from http://www.readcube.com/articles/10.1186/1471-2164-10-141

+ EIC(227.0087) Scan Exracted Solvent 3.d

Acquisition Time (min)4.3 4.32 4.34 4.36 4.38 4.4 4.42 4.44 4.46

Cou

nts 4x10

-0.250

0.250.5

0.751

1.251.5

1.752

2.252.5

2.753

3.253.5

3.75

4.377 min.4:2 FTOH - 1 Levels, 1 Levels Used, 1 Points, 1 Points Used, 2 QCs

Concentration (pg/ml)-4 -2 0 2 4 6 8 10 12 14 16 18 20 22 24 26 28 30 32 34 36 38 40 42 44 46 48 50 52 54

Res

pons

es 5x10

0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

0.9

y = 1896.210189 * x + 1.192093E-011R^2 = 1.00000000

Acquisition Time (min)4.3 4.32 4.34 4.36 4.38 4.4 4.42 4.44 4.46

2x10

-0.1

0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

0.9

1

1.1

1.2

227.0087 , 265.0253Ratio = 77.0 (97.6 %)

Peak m/z Res Abund Abund%1 580.9846 18933 411754 1002 581.9854 15792 40352 9.83 600.9901 19048 403510 984 601.9916 16046 40915 9.94

Acronym Formula Exact Mass + H Observed Mass  ppm4:2 FTOH C6H5F9O 265.0269 265.0270 ‐0.3773

6:2 FTOH C8H5F13O 365.0206 365.0206 0.00008:2 FTOH C10H5F17O 465.0142 465.0140 0.430110:2 FTOH C12H5F21O 565.0078 565.0078 0.00007:2 sFTOH C9H5F15O 415.0174 415.0190 ‐3.8553

5:1 FTOH C6H3F11O 301.0081 301.0079 0.66446:1 FTOH C7 H3 F13 O 351.0049 351.0050 ‐0.28497:1 FTOH C8 H3 F15 O 401.0017 401.0016 0.24948:1 FTOH C9 H3 F17 O 450.9985 450.9985 0.00009:1 FTOH C10 H3 F19 O 500.9953 500.9956 ‐0.598810:1 FTOH C11 H3 F21 O 550.9921 550.9922 ‐0.181511:1 FTOH C12 H3 F23 O 600.9889 600.9896 ‐1.1647MeFOSE C11 H8 F17 N O3 S 558.0026 558.0042 ‐2.8674EtFOSE C12H10F17NO3S 572.0183 572.0167 2.7971

No IRM (Lock Mass)

Acronym Precursor m/z CE Transition m/z ppm Loss10:2 FTOH 565 10V 526.9918 ‐1.3215 ‐H2O, ‐HF

F

F

F F

F

F

F

F

F

OH2+

-F2

F

F

F F

F

F

F

OH2+

CH2+

F

F

F F

F

F

F F-H2O -HF

Exact Mass: 265.0269

Exact Mass: 38.0168Exact Mass: 227.0102

Exact Mass: 227.0301

C+

F

F

F F

F

F

F F

Exact Mass: 37.9968

Acronym Observed Base Peak m/z Molecular ion ‐F2 m/z ppm Molecular ion ‐H20, ‐HF m/z ppm4:2 FTOH 227.0104 227.0301 86.7726 227.0102 ‐0.8810

Exposure to, Air Pollution, pesticides, iron Ties Autism to influences in the womb

H. M. Skip Kingston, PhDProfessor of Analytical and Bioanalytical ChemistryBayer School of Natural and Environmental SciencesDuquesne University

Scott Faber, MDDevelopmental PediatricianThe Children’s InstituteThe Children’s Institute/Duquesne University Autism Study Team

> 300K data points

Automobile Exhaust

Solvents

Flame Retardants (PBDE)

Pesticides

Phthalate

Aroclors

Percentage of children tested who displayed detectable levels of each compound

0 20 40 60 80 100

XylenePCB 28

PBDE 99PBDE 47

PCB 52PCB 138

ChlopyrifosPCB 153Benzene

PCEHexane

DEHPAcetachlor

MetalochlorPendimethalin

Toluene

0.00001

0.0001

0.001

0.01

0.1

1

10lo

lg (c

once

ntra

tion)

ug/

g

POP Organic Toxin Concentration Profile for a Father and Son

Dr. Berger Ben BergerFather Son

Exposomics is:◦ A nascent field ripe with opportunity◦ Interdisciplinary / multi-technique◦ Bioinformatics

Exposomic markets:◦ Epidemiology, pathology, “omics,” integrated biology,

clinical chemistry, toxicology, environmental, food safety, pharma & biopharma, public health…

Exposomic assays:◦ Urinary organic acids, VOA, amino acids, fatty acids,

lipids, eicosanoids from n-3 fatty acids, emerging contaminates, endocrine disruptors, fluorotelomer alcohols, estrogens & androgens, prostaglandins, acylglycines, α-keto acids, isotopomer flux, kynurenines…

May 1, 2013 (Vol. 33, No. 9)

Exposing the Exposome

Autism data◦ H. M. Skip Kingston, PhD. Duquesne University◦ Scott Faber, MD. The Children’s Institute

Agilent◦ Tim Conjelko, Jim Yano, Monty Benefiel

Johns Hopkins◦ Gabriele V. Ronnett, MD, PhD◦ David R. Graham, PhD

More, if time permits◦ The G x E relationship◦ Current studies & funding

Genetics Exposure

∝ …

China: National Basic Research Program of China J. Trans. Med. 10:A41 (2012)

EC: €17.3 million, Four year study. Nature 491:647 (2012)

UK Biobank: Long-term follow-up of health (500,000 people ages 40 –69US: CDC, NIOSH, NIEHS, EPA,

FDA

May 24, 2013: $4M Grant Awarded to Emory University, Georgia Tech to

Create Exposome Center

The Phenome Center:Imperial College and

King's College

Lead Scientist Institution NameJoseph Beckman Oregon State University Reducing Susceptibility to Environmental Stress

Throughout the Life SpanMax Costa New York University School of Medicine Research in Environmental Health SciencesDouglas Dockery Harvard University School of Public Health HSPH-NIEHS Center for Environmental HealthDavid Eaton University of Washington Center for Ecogenetics and Environmental HealthCornelius Elferink University of Texas Medical Branch at Galveston Cellular Response Mechanisms to Environmental

ChallengeThomas Gasiewicz University of Rochester Environmental Agents as Modulators of Disease

ProcessesFrank Gilliland University of Southern California Environmental Exposures, Host Factors and

Human DiseaseJohn Groopman Johns Hopkins University Johns Hopkins Center in Urban Environmental

HealthMichael Aschner Vanderbilt University Center in Molecular ToxicologyShuk-mei Ho University of Cincinnati Center for Environmental GeneticsHoward Hu University of Michigan at Ann Arbor Lifestage Exposures and Adult DiseaseDavid Johnson University of Texas MD Anderson Cancer Center Mechanisms and Prevention of Environmental

DiseaseSerrine Lau University of Arizona Southwest Environmental Health Sciences CenterTrevor Penning University of Pennsylvania Center of Excellence in Environmental ToxicologyDavid Petering University of Wisconsin Milwaukee Children's Environmental Health Sciences Core

CenterJohn Essigman Massachusetts Institute of Technology MIT Center for Environmental Health SciencesRegina Santella Columbia University Health Sciences Center for Environmental Health in Northern

ManhattanJames Swenberg University of North Carolina Chapel Hill UNC-CH Center for Environmental Health and

SusceptibilityPeter Thorne University of Iowa Environmental Health Sciences Research CenterHelmut Zarbl University of Medicine and Dentistry of New Jersey-Robert Wood

Johnson Medical SchoolResearch Center in Environmental