1. Introduction
Methods
Results
Prospects
Using Microarrays to Monitor Gene
Expression Induced by Outdoor Airborne
Particulate Matter
B. Rey de Castro, Sc.D.
Westat
Rockville, Maryland USA
March 25, 2010
reyDecastro@westat.com Genomic Exposure Assessment @ Gradient

2. Introduction
Methods
Results
Prospects
Outline
1 Introduction
2 Methods
3 Results
4 Prospects
reyDecastro@westat.com Genomic Exposure Assessment @ Gradient

3. Introduction
Methods
Results
Prospects
Outline
1 Introduction
2 Methods
3 Results
4 Prospects
reyDecastro@westat.com Genomic Exposure Assessment @ Gradient

4. Introduction
Methods
Results
Prospects
Genomic Exposure Assessment
reyDecastro@westat.com Genomic Exposure Assessment @ Gradient

5. Introduction
Methods
Results
Prospects
Particulate Matter
PM2.5: ≤ 2.5µm aerodynamic diameter
”Fine” fraction
Carbonaceous combustion by-product
Deeply respirable
Vehicle for adsorbed pollutants
Organics, metals
Health effects
Cause?
reyDecastro@westat.com Genomic Exposure Assessment @ Gradient

6. Introduction
Methods
Results
Prospects
Motivation
reyDecastro@westat.com Genomic Exposure Assessment @ Gradient

7. Introduction
Methods
Results
Prospects
Aims
Hypothesis
The organic fraction of PM2.5 alters expression of
genes related to detoxification, stress response,
inflammatory response, apoptosis in normal human
epithelial cells of the small airway
Simple pilot project
Observe gene expression profiles induced by PM2.5
collected in Boston
Identify groups of co-expressed genes
Correlate gene expression profiles with season,
temperature, dew point, PM mass, PAH concentration
profile
reyDecastro@westat.com Genomic Exposure Assessment @ Gradient

8. Introduction
Methods
Results
Prospects
Outline
1 Introduction
2 Methods
3 Results
4 Prospects
reyDecastro@westat.com Genomic Exposure Assessment @ Gradient

9. Introduction
Methods
Results
Prospects
High-Volume Cascade Impactor
reyDecastro@westat.com Genomic Exposure Assessment @ Gradient

10. Introduction
Methods
Results
Prospects
PUF Sample
reyDecastro@westat.com Genomic Exposure Assessment @ Gradient

11. Introduction
Methods
Results
Prospects
Small Airway Epithelial Cells (SAEC)
reyDecastro@westat.com Genomic Exposure Assessment @ Gradient

12. Introduction
Methods
Results
Prospects
Design Overview
reyDecastro@westat.com Genomic Exposure Assessment @ Gradient

13. Introduction
Methods
Results
Prospects
DNA Microarray
reyDecastro@westat.com Genomic Exposure Assessment @ Gradient

14. Introduction
Methods
Results
Prospects
Outline
1 Introduction
2 Methods
3 Results
4 Prospects
reyDecastro@westat.com Genomic Exposure Assessment @ Gradient

15. Introduction
Methods
Results
Prospects
Summer vs. Field Blank: SAMs RMA
reyDecastro@westat.com Genomic Exposure Assessment @ Gradient

16. Introduction
Methods
Results
Prospects
Cytochrome P450 Genes
Gene Fold-Change 90 % CI
CYP1B1 (dioxin-inducible) 4.18 [3.24 : 5.60]
CYP1A1 (aromatic-inducible) 3.88 [2.84 : 5.39]
CYP2C9 1.36 [1.14 : 1.60]
reyDecastro@westat.com Genomic Exposure Assessment @ Gradient

17. Introduction
Methods
Results
Prospects
Inflammation Genes
Gene Fold-Change 90 % CI
interleukin-1β 1.63 [1.48 : 1.80]
calgranulin A 1.62 [1.28 : 2.08]
interleukin-1 receptor antagonist 1.50 [1.34 : 1.70]
calgranulin B 1.48 [1.22 : 1.79]
interleukin-8 -1.64 [-1.17 : -2.25]
reyDecastro@westat.com Genomic Exposure Assessment @ Gradient

18. Introduction
Methods
Results
Prospects
Summary: Gene Expression Response
Scope: 100 genes
Detoxification: strongest
Increased CYP1B1, CYP1A1, CYP2C9
Inflammation: moderate
Increased interleukin-1β, calgranulins A and B
Increased interleukin-1 receptor antagonist
Decreased interleukin-8
reyDecastro@westat.com Genomic Exposure Assessment @ Gradient

19. Introduction
Methods
Results
Prospects
Outline
1 Introduction
2 Methods
3 Results
4 Prospects
reyDecastro@westat.com Genomic Exposure Assessment @ Gradient

20. Introduction
Methods
Results
Prospects
Genomic Exposure Assessment
Correlate gene expression with PM exposure
When, Where
Sources
Constituents
Other phenomena
Genes-Environment interactions
Airborne biologic agents?
Surveillance
reyDecastro@westat.com Genomic Exposure Assessment @ Gradient

21. Introduction
Methods
Results
Prospects
Genomic Exposure Assessment
reyDecastro@westat.com Genomic Exposure Assessment @ Gradient

22. Introduction
Methods
Results
Prospects
Contact
On SlideShare: http://cli.gs/microarrayGradient
B. Rey de Castro, Sc.D.
410-929-3583
reyDecastro@westat.com Genomic Exposure Assessment @ Gradient