Craniofacial Dysmorphism & Fetal Alcohol Exposure Tatiana Foroud and Peter Hammond (PIs) Leah Wetherill, Mike Suttie

Update on Image Collection Site As of Jan 2013 Images (# subjects)1 As of April 2014 Images As of February 2015 Images San Diego 239 (215) 349 (273) 416 (312) UCLA/USC 52 (52) 75 (67) 104 (88) Atlanta 147 (147) 203 (203) 248 (248) Minneapolis 50 (50) 174 (139) 238 (236) Ukraine 47 (34) 159 (95) South Africa (Jacobson) 223 (223) 517 (312) South Africa (May) 37 (37) South Africa (PASS) 1,738 (1,221) 2,361 (1,221) Totals 2,533 (1,979) 3,763 (2,286) 4,080 (2,549) 1 Some subjects have longitudinal image collection

Racial Distribution of Subjects Alcohol Exposure

Update on Saliva Collection Site As of Jan 2013 As of April 2014 As of Feb 2015 San Diego 131 287 421 (246) UCLA/USC 45 110 159 (108) Atlanta 28 194 353 (227) Minneapolis 170 349 (194) Ukraine 01 South Africa (Jacobson) 225 South Africa (PASS) 1 Collection of samples done as part of parent protocol

Use of DNA for Research GWAS completed in 240 individuals 700,000 SNPs genotyped (Illumina OmniExpress) Variable alcohol exposure and phenotype Data used for candidate gene studies Genotype x alcohol interaction Developmental Project completing GWAS in another 327 subjects 2 million SNPs being genotyped (Illumina MEGA)

FYN philtrum pc2 philtrum pc9 executive function CC mid posterior

New Directions Develop multidimensional phenotypes that can be used for analysis Focus of the RSA symposium in June 2015

Analysis Objectives Develop a screening tool that would utilize the data from the 3D facial images and could be widely used to accurately identify individuals with a high likelihood of alcohol exposure Recruit & analyze facial imaging data from very young populations to develop a screening tool that accurately identifies high risk individuals for future intervention Combine face images, neurobehavioral data & brain images to identify common pathways & improve diagnosis of prenatal alcohol exposure Extend existing and develop novel techniques and associated software to cope with demands of larger datasets and more diverse comparison of controls, alcohol exposed and other developmentally delayed subjects while accommodating multiple anatomical images per subject Extend preliminary genetic studies through collection of DNA samples for new subjects and focused analysis to replicate candidate genes identified in basic science components.

COMPLETED TATIANA Analysis Objectives Develop a screening tool that would utilize the data from the 3D facial images and could be widely used to accurately identify individuals with a high likelihood of alcohol exposure Recruit & analyze facial imaging data from very young populations to develop a screening tool that accurately identifies high risk individuals for future intervention Combine face images, neurobehavioral data & brain images to identify common pathways & improve diagnosis of prenatal alcohol exposure Extend existing and develop novel techniques and associated software to cope with demands of larger datasets and more diverse comparison of controls, alcohol exposed and other developmentally delayed subjects while accommodating multiple anatomical images per subject Extend preliminary genetic studies through collection of DNA samples for new subjects and focused analysis to replicate candidate genes identified in basic science components. COMPLETED TATIANA

Face–cognitive impairment in exposed children not diagnosable as FAS/PFAS HE (Suttie et al, Pediatrics, 2013)

HE1FAS-like HE2 CTRL like Face–cognitive impairment in exposed children not diagnosable as FAS/PFAS (Suttie et al, Pediatrics, 2013) HE1 FAS WISC IQ 65.5 65.4 CVLT-C1 40.0 42.7 CVLT-C2 84.3 88.5 HE2 HC WISC IQ 73.3 CVLT-C1 47.3 45.8 CVLT-C2 93.7 93.2 HE1FAS-like HE2 CTRL like

N=192 (JACOBSEN) Cape- Coloured N=227 (CIFASD) Caucasian Recapitulate Pediatrics paper analysis from Cape Coloured to CIFASD Caucasian Suttie et al (2013) Pediatrics, 131 (3), e779-e788 HC = 69 FAS = 22 HE = 75 HC = 69 FAS/PFAS = 48 HE = 75 N=192 (JACOBSEN) 5yr-15 yr Cape- Coloured HC = 121 FAS = 34 HE = 74 N=227 (CIFASD) 5yr-15+ yr Caucasian

Does HE face predict cognitive impairment ? CAPE CAUCASIAN + HISPANIC FAS HE FAS-like HE control-like

Does HE face predict cognitive impairment ? CAPE CAUCASIAN ONLY FAS HE FAS-like HE control-like

Probability of agreeing with clinical diagnosis of pair of faces – 1 HC & 1 FAS HC vs FAS CAPE   CAUCASIAN CM LDA SVM PROFILE 0.97 0.83 0.85 0.88 FACE 0.95 0.87 MALAR 0.79 0.80 NOSE 0.90 0.92 0.91 0.94 PHILTRUM 0.72 0.78 0.86 Cape FAS Caucasian FAS

Facial growth reduction due to prenatal alcohol exposure (PAE): Cape >> Caucasian

Homogeneity of PAE induced facial dysmorphism : Cape FAS > Caucasian FAS

Mean curvature at point on face surface (not normalised) “concave” “flat” “convex”

Groove & curl at point on philtrum   subnasale lip centre CURL GROOVE

The philtrum: the curl and groove of it low groove/high curl high groove/high curl/ CURL low groove/low curl high groove/low curl GROOVE

Variation in philtrum groove & pillar definition ). Variation in philtrum groove & pillar definition

Mean groove & philtral pillar definition : Cape Control << Caucasian Control CONTROL shape curl groove C A P E C A U

PAE induced effect on philtrum groove/pillars: Cape FAS << Caucasian FAS CONTROL FAS shape curl groove shape curl groove C A P E C A U

Cape vs Caucasian differences explaining discrimination variation with location FAS facial growth delay: Cape > Caucasian FAS facial dysmorphism: Cape more homogeneous HC upper philtrum smoothness: Cape > Caucasian FAS philtrum smoothness: Caucasian > Cape (?) FAS malar region effect: Cape > Caucasian CIFASD Hispanic & African-American analysis to be completed

Measuring volume of philtral groove facet on philtrum facet on lid positive voxel projection plane negative voxel curl

PHILTRUM VOLUME vs AGE: Caucasian controls

Tentative Measure of Philtrum Smoothness “Philtrum Groove Index”   subnasale lip centre Philtrum Groove Index = Philtrum Volume / Philtrum Length corresponds to average cross sectional area

CAUCASIAN:Philtrum Groove Index vs PFL (HC=45; FAS=15; HE=40) * Pearson coeff 0.315, p=0.035 for controls

CAPE:Philtrum Groove Index vs PFL (HC= 64; FAS=17 ) * Pearson coeff 0.356, p < 0.01 for controls

Face screening tool TOOL

3dMD camera vs Canfield H1 hand-held

3dMD CCA386 H1 CCA386

H1 CCA386 3DMD CCA386 H1 vs 3DMD means H1 MEAN SIG 3DMD MEAN SIG

Philtrum Volume 3dMD vs H1

Mean FAS (N=12) vs Mean HC (N=21) Face & CC and Philtrum & CC

Profile+CC closest mean classification Vs Wiscfullscalecomp MEAN HC MEAN FAS

Correlation of Philtrum Groove Index & Corpus Callosum PCA Modes Pearson 0.381** 0.363* 0.349* Sig (2-tailed) 0.010 0.014 0.019 Mode 9 1% variance Mode 29 0.06% variance Mode 2 14.2% variance

Papers Cape – Caucasian differences - DRAFT Screening tool – DRAFT USER DOC Prenatal with PASS – CIFASD report close to draft Face-cognition-brain – DRAFT end of April

CIFASD IV – Future Face Work Automated Landmarking (using 2D and 3D) Automated Analysis with Cloud based models Tablet/Smartphone 2D and 3D image capture Selected PCA modes per individual for accurate face synthesis in dense surface models Non-Caucasian and Ad-mix populations

Analysis of 21 3D U/S : Alison Noble (Oxford) Tom Rackham extracted profiles Shape analysis – PC1 PC1 correlates with Trimester 2 drinking (DM-Stat/PASS) Not adjusted for Gest Age Adjusted for Gest Age drinks per drinking day by trimester and up to date of measurement r=0.42, p=0.0597 r=0.42, p=0.0667 binge vs. no binge by trimester and up to date of measurement (t-test) p=0.031* drinks per drinking week by trimester and up to date of measurement r=0.52, p=0.017* r=0.52, p=0.019*