Oral microbiome development & early childhood caries:

a case-control study
Freida Blostein1; Deesha Bhaumik1; Elizabeth Salzman1; Dr. Daniel McNeil2;
Dr. Mary Marazita2; Dr. Betsy Foxman1
1 University of Michigan School of Public Health Department of Epidemiology
2 Center for Oral Health Research in Appalachia West Virginia University School of Dentistry

1
Background: Ecologic model of dental caries Important findings & significance
Early childhood caries (ECC): multifactorial disease
catalyzed by oral bacteria. - Co-occurring taxa are strongly correlated with age and dental decay and less strongly correlated with pH, sugared-beverage consumption  Are
host genetics and immunity more important risk factors?  Next steps: incorporate host genomic information.
Old model: S. mutans  disease
- Low abundance, non-acidogenic taxa which play important roles in biofilm structure were network hubs.  Can these taxa be targeted for
New model: Structure and function of the microbial intervention at critical periods?  Next steps: examine how hub bacteria influence ecologic succession of oral cavity over time.
community as whole  dysbiosis and disease.

Image from Pitt et al1

Methods Results
Study design & study population Figure 2: Modules correlate with host characteristics including tooth decay
Prenatal visit 2 months First tooth 1 year 2 years 3 years 4 years 5 years

(+) Correlation

- Subsample of Center for Oral Health Research in Appalachia (COHRA) cohort: N=189 Caucasian children
- At regular dental exams exam, saliva samples taken. Telephone interviews with parents about: child’s dental hygiene, child’s diet, and
sociodemographic characteristics.

Bioinformatics & network analysis

0.100

0.075

(-) Correlation

Relative Abundance
V4 region Mothur: Sequences cleaned MED: Sequences binned to Network analysis: Performed in 0.050

16S rRNA oligotypes WGCNA

- Network analysis: Correlation between oligotypes used to calculate topological overlap matrix (TOM). Oligotypes clustered by TOM dissimilarity. 0.025
Figure 3: Hub taxa may be related to biofilm formation
- Dynamic tree cut to extract clusters of co-occurring oligotypes. Modules correlated with sample characteristics and network hubs identified. 3a: Turquoise module hub Cornyebacterium matruchotti has higher relative abundance in those with dental decay & in later time
0.000
points; forms “corn-cob”
~0 ~structures
3 ~ 9 in plaque
~ 12 ~ 24 ~ 37 ~ 49 ~ 60
Average Age (Months)

0.100 as.factor(CurrentCase) Current case Eventual case Never case

Results
Figure 1: Genus composition of co-occurring taxa clusters (‘modules’) identified from network analysis 0.075
Relative Abundance

0.100
0.050

0.075
Relative Abundance

0.025

0.050

0.000
Image from Mark Welch et al2 : . Fluorescence in situ
~0 ~3 ~9 ~ 12 ~ 24 ~ 37 ~ 49 ~ 60
hybridization, Cornyebacterium matruchotti shown in pink
0.025 Average Age (Months) with cocci bacteria in green
3b Blue module hub Current
Fusobacterium
case periodonticum
Eventual case Never case
as.factor(CurrentCase) 3c Fusobacterium oligotypes assigned to
has higher
0.000relative abundance in those without different modules: may reflect strain preference
dental decay ~ 0 ~3 ~9 ~ 12 ~ 24
Average Age (Months)
~ 37 ~ 49
for soft tissues vs teeth3
~ 60

as.factor(CurrentCase) Current case Eventual case Never case

0.10
Relative Abundance

0.05

*S. mutans was assigned to the Brown module 0.00

~0 ~3 ~9 ~ 12 ~ 24 ~ 37 ~ 49 ~ 60
Average Age (Months)

References as.factor(CurrentCase) Current case Eventual case

Acknowledgements
Never case

1Pitts NB, Zero DT, Marsh PD, et al. Dental caries. Nat Rev Dis Prim. 2017;3(May):17030. doi:10.1038/nrdp.2017.30.
2Mark Welch JL, Rossetti BJ, Rieken CW, Dewhirst FE, Borisy GG. Biogeography of a human oral microbiome at the micron scale. Proc Natl Acad Sci U S A.
Mentor: Dr. Betsy Foxman
2016;113(6):E791–E800. doi:10.1073/pnas.1522149113
3Eren AM, Borisy GG, Huse SM, Mark Welch JL. Oligotyping analysis of the human oral microbiome. Proc Natl Acad Sci. 2014;111(28):E2875 LP-E2884.
Sources of Funding: T32 HG00040 (Trainee grant); R01DE014899 (COHRA grant; PI Dr. Bets
doi:10.1073/pnas.1409644111. Foxman & Dr. Mary Marazita)