Abstract
Oral feeding competency is a major determinant of length of stay in the neonatal intensive care unit. An infant must be able to consistently demonstrate the ability to take all required enteral nutrition by mouth before discharge home. Most infants born prematurely (<37 weeks) will require days, if not weeks, to master this oral feeding competency skill. Inappropriately timed feeding attempts can lead to acute and long-term morbidities, prolonged hospitalizations, and increased health-care costs. Previously, a panel of five genes involved in essential developmental pathways including sensory integration (nephronophthisis 4, Plexin A1), hunger signaling [neuropeptide Y2 receptor (NPY2R), adenosine-monophosphate-activated protein kinase (AMPK)], and facial development (wingless-type MMTV integration site family, member 3) required for oral feeding success were identified in neonatal saliva. This study aimed to translate these five transcriptomic biomarkers into a rapid proteomic platform to provide objective, real-time assessment of oral feeding skills, to better inform care, and to improve neonatal outcomes. Total protein was extracted from saliva of 10 feeding-successful and 10 feeding-unsuccessful infants matched for age, sex, and post-conceptional age. Development of immunoassays was attempted for five oral feeding biomarkers and two reference biomarkers (GAPDH and YWHAZ) to normalize for starting protein concentrations. Normalized protein concentrations were correlated to both feeding status at time of sample collection and previously described gene expression profiles. Only the reference proteins and those involved in hunger signaling were detected in neonatal saliva at measurable levels. Expression patterns for NPY2R and AMPK correlated with the gene expression patterns previously seen between successful and unsuccessful feeders and predicted feeding outcome. Salivary proteins associated with hunger signaling are readily quantifiable in neonatal saliva and may be utilized to assess oral feeding readiness in the newborn. This study lays the foundation for the development of an informative, rapid, proteomic platform to assess neonatal oral feeding maturation.
Highlights
Oral feeding competency is a discharge requirement from the neonatal intensive care unit (NICU), there is currently a paucity of objective assessment tools to determine oral feeding maturity in this population [1, 2]
Salivary gene expression analyses on hundreds of premature infants at both pre- and post-oral feeding success, identified five genes involved in oral feeding maturity that were differentially expressed between successful and unsuccessful oral feeders [12]
No compatible antibody pairs were found for nephronophthisis 4 (NPHP4) and Plexin A1 (PLXNA1)
Summary
Oral feeding competency is a discharge requirement from the neonatal intensive care unit (NICU), there is currently a paucity of objective assessment tools to determine oral feeding maturity in this population [1, 2]. Salivary gene expression analyses on hundreds of premature infants at both pre- and post-oral feeding success, identified five genes involved in oral feeding maturity that were differentially expressed between successful and unsuccessful oral feeders [12] These genes included Plexin A1 (PLXNA1), neuropeptide Y2 receptor (NPY2R), adenosine-monophosphateactivated protein kinase (AMPK), wingless-type MMTV integration site family, member 3 (WNT3), and nephronophthisis 4 (NPHP4). While this prior study demonstrated the feasibility of utilizing saliva as a non-invasive biofluid to detect transcriptomic biomarkers associated with neonatal developmental milestones, the ability to monitor these markers in a timely manner to inform care remains a challenge. This study aimed to translate the previously described gene expression panel to a salivary proteomic platform to rapidly and objectively assess oral feeding readiness to limit neonatal morbidities and improve outcomes
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