Variants in the Hippo Signaling Pathway are Associated with Craniofacial Skeletal Form and BMI
Authors: Dema Agilda , Yip Kayla , Spencer Nicolette , Ro Eugene , Ehrlich Daniel , Padala Soumya , Bjork Bryan , Miller Steven 
Published: April 2020

Childhood obesity rates in the United States have demonstrated a marked increase since the late 20th century. Current research has suggested that obesity can cause increases in bone density and an acceleration of skeletal growth. While studies have shown that obesity may be associated with altered craniofacial form, the mechanism and associated factors through which this interaction operates remains poorly understood. This study explores a potential link between obesity and craniofacial skeletal form as a function of the Hippo signaling pathway, an important regulator of organ size, tissue development, and craniofacial development.

Lateral cephalograms from a sample of n=64 patients between the ages 7 to 17 were obtained from the Midwestern University Multispecialty Clinic. BMI percentile was computed for all children in the study following CDC guidelines (normal weight n=37, overweight n=9, obese n=18). 45 2D landmarks were registered via Generalized Procrustes Analysis (GPA) in order to facilitate a geometric morphometric (GM) analysis. Healthy Eating Index (HEI) scores obtained from Block Food Frequency Questionnaire (FFQ) surveys were used to control for the effects of diet in all subsequent analyses. Shape residuals were submitted to Principal Components Analysis (PCA) and Canonical Variate Analysis (CVA) to identify which aspects of skeletal form differentiate individual phenotypes based on BMI. Finally, associations between these phenotypes and 23 single nucleotide polymorphisms (SNPs) in seven genes within the Hippo signaling pathway were explored.

Principal Components (PCs) accounting for at least 1% of facial shape variation were examined in order to determine components that differentiate between normal and obese groups. Based on this, suggestive/significant differences were found in PC 8 (p=0.056) and PC12 (p=0.012) based on BMI category. PC 8 (accounting for 3.6% of the total shape variation) indicates that chin projection, nose length, and mandibular morphology differentiate between normal and obese children. SNPs in TEAD3 (p=0.04) and FAT4 (p = 0.02) were significantly associated with shape variation along PC8. PC 12 (2.6% of the total shape variation) showed significant variation in chin projection and decreased incisor size in the obese group relative to the normal weight group. Additionally, a SNP within LATS2 (p = 0.04) was significantly correlated with shape variation along PC12. Finally, results from the CVA identified two components with CV2 indicating a significant relationship with LEF1 (p= 0.02). CV2 demonstrates increased facial projection in individuals with obesity.

Across all analyses, we found a consistent pattern in mandibular length, facial projection, and incisor size that differentiated between normal and obese individuals. SNPs within key genes in the Hippo signaling pathway were associated with these phenotypes, indicating a potential causative relationship. Future studies will explore this relationship further.