Shotgun and quantitative proteomics characterization of altered carbohydrate metabolism in preterm infants

Location

Medical Education Building, LSUHSC-NO

Presentation Date

10-10-2019 10:00 AM

End Date

10-10-2019 12:00 PM

Description

noted to occur in preterm infants who, at birth, exhibit immature gut development. It is unclear whether such common conditions are associated with practices in the neonatal intensive care unit that may contribute to gastrointestinal development, such as frequency of enteral or parenteral feeding, and/or from polymorphic variation that is age- and individual-dependent. Objective: The human gut contains a dense, complex, and diverse set of proteins for digestion and for chemical communication with the resident microbiome. Our hypothesis is that enzymes involved in host carbohydrate biochemistry can undergo a biochemical switch as a function of development. Its significance is that isoform differences would underlie altered composition of metabolic intermediates in preterm infants with immature gut development would be expected to result in hyperglycemia and other medical concerns that may be age-dependent. Design, setting, and participants: In this initial presented work, we performed an exploratory proteomic analysis of human gut lumen proteins shed in stool of preterm infants who did not have any gut inflammatory or disease conditions. Preterm infants were prospectively enrolled at Children’s Hospital and Touro Infirmary. For this analysis, gestational age of infants was 26, 27, and 32 weeks; male:female ratio was 1:2. Stool from discarded diapers were stored at 4oC at hospital specimen refrigerators until couriered to research laboratory for sample processing. Exposures: Infant stool samples were collected between 24-40+ weeks postconceptual age (PCA). As this is an observational study, enrolled infants underwent testing, management, and treatment at physician and hospital site discretion. Main outcomes and measures: We used non-targeted, bottom-up mass spectrometry-based proteomics for the first deep proteome measurements of human preterm infant fecal samples at 32 weeks postconceptual age. Second, our discovery-based quantitative proteomic strategy employed tandem mass tag (TMT) labeling, in conjunction with liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS), to compare the gut lumen proteomes of 10 patient samples (from 3 patients) who were not suspected or diagnosed with gut inflammation or disease. Results: We identified 635 human proteins in the neonatal gut lumen protein profile. Of these, 65 proteins were isoforms not found in previously published adult gut proteome. Human proteins needed for carbohydrate metabolism and energy production were highly represented. Using the 10- plex TMT labeling approach, we identified proteins that have differences in abundance in the gut lumen between control infants. Conclusions: Evaluation of metabolic proteins in human development is crucial to understanding their function in health and in disease. Discovery that 10% of human proteins found in preterm infant gut are non-canonical isoforms have broad impacts in determining nutritional management in the neonatal intensive care unit.

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Mentor: Sunyoung Kim PhD (Dept. Biochemistry & Molecular Biology)

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Oct 10th, 10:00 AM Oct 10th, 12:00 PM

Shotgun and quantitative proteomics characterization of altered carbohydrate metabolism in preterm infants

Medical Education Building, LSUHSC-NO

noted to occur in preterm infants who, at birth, exhibit immature gut development. It is unclear whether such common conditions are associated with practices in the neonatal intensive care unit that may contribute to gastrointestinal development, such as frequency of enteral or parenteral feeding, and/or from polymorphic variation that is age- and individual-dependent. Objective: The human gut contains a dense, complex, and diverse set of proteins for digestion and for chemical communication with the resident microbiome. Our hypothesis is that enzymes involved in host carbohydrate biochemistry can undergo a biochemical switch as a function of development. Its significance is that isoform differences would underlie altered composition of metabolic intermediates in preterm infants with immature gut development would be expected to result in hyperglycemia and other medical concerns that may be age-dependent. Design, setting, and participants: In this initial presented work, we performed an exploratory proteomic analysis of human gut lumen proteins shed in stool of preterm infants who did not have any gut inflammatory or disease conditions. Preterm infants were prospectively enrolled at Children’s Hospital and Touro Infirmary. For this analysis, gestational age of infants was 26, 27, and 32 weeks; male:female ratio was 1:2. Stool from discarded diapers were stored at 4oC at hospital specimen refrigerators until couriered to research laboratory for sample processing. Exposures: Infant stool samples were collected between 24-40+ weeks postconceptual age (PCA). As this is an observational study, enrolled infants underwent testing, management, and treatment at physician and hospital site discretion. Main outcomes and measures: We used non-targeted, bottom-up mass spectrometry-based proteomics for the first deep proteome measurements of human preterm infant fecal samples at 32 weeks postconceptual age. Second, our discovery-based quantitative proteomic strategy employed tandem mass tag (TMT) labeling, in conjunction with liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS), to compare the gut lumen proteomes of 10 patient samples (from 3 patients) who were not suspected or diagnosed with gut inflammation or disease. Results: We identified 635 human proteins in the neonatal gut lumen protein profile. Of these, 65 proteins were isoforms not found in previously published adult gut proteome. Human proteins needed for carbohydrate metabolism and energy production were highly represented. Using the 10- plex TMT labeling approach, we identified proteins that have differences in abundance in the gut lumen between control infants. Conclusions: Evaluation of metabolic proteins in human development is crucial to understanding their function in health and in disease. Discovery that 10% of human proteins found in preterm infant gut are non-canonical isoforms have broad impacts in determining nutritional management in the neonatal intensive care unit.