1. Diagnostic / Prognostic
CIFASD Strategic Objective: ‘Clinically-useful diagnostic criteria for FASD’
FACIAL MORPHOLOGY
Dysmorphology Core
3D Camera Study
BRAIN STRUCTURE
sMRI / DTI
NEUROBEHAVIOR
“BIOMARKERS”
Diagnostic / Prognostic
BRAIN FUNCTION
fMRI / MEG 3

2. Biomarkers of Ethanol Exposure / Effect?
MCV, GGT, CDT ( blood )
FAEEs ( meconium, hair? )
Ethyl Glucuronide ( blood, urine, hair? )
“Novel Biomarkers” ( blood?, placenta? ) Genomics Proteomics Metabolomics microRNAs 3

3. Prenatal Ethanol Exposure Paradigm
VOLUNTARY DRINKING
- 5% Ethanol in 0.066% saccharin water
- Two-bottle choice paradigm
- Intermittent consumption (4 hr/day)
- Peak maternal serum EtOH: Long-Evans rats: mg / dL Black 6/J mice: mg / dL
- Saccharin water, volume-matched control 3

4. Altered Gene Expression
Cannabinoid Receptor S100 Calcium Binding Protein A4
Galanin Receptor Integrin Alpha 7
Gremlin S100 Calcium Binding Protein G
Hemoglobin Epsilon Secreted Frizzle-related Protein 4
Matrix Metalloproteinase 2 S100 Calcium Binding Protein B
Deiodinase Iodothyronine 2 Lipopolysaccharide Binding Protein
Galanin Hemoglobin Delta
Toll-like Receptor Placental Growth Factor
Transforming Growth Factor Alpha
Heart & Neural Crest Derivative 2
11b-Hydroxysteroid Dehydrogenase 2
Transient Receptor Potential Cation Channel 4
Serine Peptidase Inhibitor, Kazal Type 5
EGF-containing Fibulin-like Extracellular Matrix Protein 1 3

5. 2D PAGE Analysis of Placental Proteins – 150 mg samples
Control
Ethanol
 pH  mw 
Courtesy of Prof. Thierry Jouenne & colleagues, University of Rouen

6. Role of Animal Models in Biomarker Development
Rodents - Rapid, cost-effective identification of candidates - Ethanol-dose, timing, frequency characterizations - Other “risk factor” interaction assessments (nicotine, stress, malnutrition, heavy metals, etc.) - Correlation with behavioral / physiologic outcomes
Sheep Model - Validation of biosignature patterns - Functional Implications 3

7. Biomarker Development Strategy
Preclinical Studies Clinical Studies
Identification Maternal Data & Tissue Banking
Characterization Offspring Measures
Validation Biomarker Assessment 3

8. Consensus Protocol Information / Sample Collection
Interview: TWEAK, AUDIT, Maternal Information
Maternal: Blood, Urine, Hair Ultrasound (including Doppler) Placenta, Cord Blood
Neonate: Blood, Meconium, Hair? Neurobehavioral Data 3

9. Consensus Protocol Information / Sample Collection Times
First Prenatal Visit: (1st trimester)
Follow-up Visit: (3rd trimester)
Delivery:
Follow-up: (6 to 12 months of age) 3

10. Prospective Human Biomarker Study Sites
PASS
France
CIFASD - Ukraine SDSU/UCSD, UCLA, Emory, UNM, others ?
Other International Sites ? South Africa Finland Italy Canada Sweden
National Children’s Study New Mexico, California, Maryland, New York, others? 3

13. Decreased Protein Expression
Altered Gene Expression
Cannabinoid Receptor S100 Calcium Binding Protein A4
Galanin Receptor Integrin Alpha 7
Gremlin S100 Calcium Binding Protein G
Hemoglobin Epsilon Secreted Frizzle-related Protein 4
Matrix Metalloproteinase S100 Calcium Binding Protein B
Deiodinase Iodothyronine Lipopolysaccharide Binding Protein
Heart & Neural Crest Derivative 2 Transient Receptor Potential Cation Channel 4
Galanin Serine Peptidase Inhibitor, Kazal Type 5
Hemoglobin Delta Toll-like Receptor 4
Transforming Growth Factor Alpha Placental Growth Factor
11b-Hydroxysteroid Dehydrogenase 2
EGF-containing Fibulin-like Extracellular Matrix Protein 1
Decreased Protein Expression
Proteasome Subunit Beta Type 3 Adenine Phosphoribosyl Transferase
Keratin Desmin
Enolase Alpha Valosin-containing Protein
Protein Disulfide Isomerase Eukaryotic Translation Elongation Factor 2
Stomatin Peroxiredoxin 5 Precursor
Chaperon Protein TCP1 Subunit Hydroxyisobutyrate Dehydrogenase 3

14. Long-Term Strategy Preclinical Clinical Biomarkers Biomarkers
Fetal / Neonatal
Brain Effects
Weanling
Behaviors &
Neurophysiology
Neonatal
Behaviors &
Neurophysiology 3