Computational Gene Expression Modeling Identifies Salivary Biomarker Analysis that Predict Oral Feeding Readiness in the Newborn

Abstract

Objective-To combine mathematical modeling of salivary gene expression microarray data and systems biology annotation with RT-qPCR amplification to identify (phase I) and validate (phase II) salivary biomarker analysis for the prediction of oral feeding readiness in preterm infants.Study design-Comparative whole transcriptome microarray analysis from 12 preterm newborns pre-and post-oral feeding success was used for computational modeling and systems biology analysis to identify potential salivary transcripts associated with oral feeding success (phase I). Selected gene expression biomarkers (15 from computational modeling; 6 evidencebased; and 3 reference) were evaluated by RT-qPCR amplification on 400 salivary samples from successful (n=200) and unsuccessful (n=200) oral feeders (phase II). Genes, alone and in combination, were evaluated by a multivariate analysis controlling for sex and post-conceptional age (PCA) to determine the probability that newborns achieved successful oral feeding.Results-Advancing post-conceptional age (p < 0.001) and female sex (p = 0.05) positively predicted an infant's ability to feed orally. A combination of five genes, NPY2R (hunger signaling), AMPK (energy homeostasis), PLXNA1 (olfactory neurogenesis), NPHP4 (visual behavior) and WNT3 (facial development), in addition to PCA and sex, demonstrated good accuracy for determining feeding success (AUROC = 0.78). Publisher's Disclaimer: This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final citable form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.The authors declare no conflicts of interest. Conclusions-We have identified objective and biologically relevant salivary biomarkers that noninvasively assess a newborn's developing brain, sensory and facial development as they relate to oral feeding success. Understanding the mechanisms that underlie the development of oral feeding readiness through translational and computational methods may improve clinical decision making while decreasing morbidities and health care costs. NIH Public AccessPreterm births affect an estimated 11.5% of all pregnancies in the United States resulting in medical costs exceeding $26 billion annually 1 . Prior to discharge, each infant must demonstrate mature oral feeding skills in accordance to the American Academy of Pediatrics' guidelines 2 . The determination of oral feeding readiness in the preterm newborn remains a significant clinical challenge 3 . Oral feeding is a complex developmental task requiring maturation and integration of the nervous, gastrointestinal, sensory, skeletal muscular and hypothalamic systems 4 . Disruption or delayed maturation in one or several of these developmental syst...

Pediatric Feeding/Swallowing: Yesterday, Today, and Tomorrow

“…43,44 Another area under investigation involves the use of objective salivary biomarkers related to the neonate's developing brain and sensory and facial development in relation to oral feeding success. 45 In the future, these biomarkers may enable clinicians to determine when brain maturation supports the initiation of oral feeding and may increase our understanding of the mechanisms in the developing brain that underlie oral feeding readiness. 45 Investigations are needed to determine the long-term outcomes associated with feeding interventions as new technologies emerge.…”

“…45 In the future, these biomarkers may enable clinicians to determine when brain maturation supports the initiation of oral feeding and may increase our understanding of the mechanisms in the developing brain that underlie oral feeding readiness. 45 Investigations are needed to determine the long-term outcomes associated with feeding interventions as new technologies emerge. Development of objective and reproducible noninvasive measures that enhance the clinical assessment is desperately needed.…”

Oral colostrum priming shortens hospitalization without changing the immunomicrobial milieu

“…Breast milk feeding is associated with an altered oral and nasopharyngeal microbiota later in infancy 6, 7 . However, oral feeding is not practical in most preterm infants due to developmental immaturity 8 . Recently, the practice of early (as soon as produced by the mother) oral administration of mother’s own colostrum was proposed as a protective strategy 9 .…”

The Neonatal Salivary Transcriptome

“…Current standards of care mandate the use of cue-based feeding protocols that ultimately lack the necessary specificity and sensitivity required to avoid placing infants at risk for choking, desaturation, and feeding aversion. Prior research has shown that for a newborn to successfully feed they must integrate numerous cranial nerves and muscles, along with their sensory, neurodevelopmental, and hunger signaling pathways (Barlow 2009;Maron et al 2015). Interestingly, in the original neonatal salivary transcriptome study, computational and systems biology reviews of gene targets highlighted biological pathways associated with feeding maturity.…”

“…Computational analysis and systems biology review of the microarray data identified a panel of genes (n ¼ 24) that were involved in the diverse biological systems required in oral feeding maturation including sensory integration, facial development, neurodevelopment, hunger signaling, and digestive system development (Maron et al 2015). A large prospective validation phase testing each of these biomarkers, alone and in combination, on 400 saliva samples resulted in the further identification of five genes (NPY2R, WNT3, PLXNA1, NPHP4, and AMPK), that when combined with postconceptional age and sex had good accuracy at predicting feeding success in the newborn (area under the receiving operator curve ¼ 0.78).…”