Neonatal Pulmonology 3: BPD Clinical and Translational
277 - Whole Lung Proteomics in Bronchopulmonary Dysplasia
Sunday, April 30, 2023
3:30 PM – 6:00 PM ET
Poster Number: 277 Publication Number: 277.341
Andrew M. Dylag, University of Rochester, Rochester, NY, United States; Ravi Misra, Golisano Children's Hospital at The University of Rochester Medical Center, Rochester, NY, United States; Gautam Bandyopadhyay, University of Rochester School of Medicine and Dentistry, Rochester, NY, United States; Cory Poole, University of Rochester, Tarrytown, NY, United States; Heidie L. Huyck, URMC, Rochester, NY, United States; JEANNIE HAAK, UNIVERSITY OF ROCHESTER, ROCHESTER, NY, United States; Gail Deutsch, University of Washington School of Medicine, Seattle, WA, United States; Heather M. Olson, PNNL, Richland, WA, United States; Philip J. Katzman, University of Rochester School of Medicine and Dentistry, Rochester, NY, United States; Jeffrey M. Purkerson, University of Rochester School of Medicine and Dentistry, Rochester, NY, United States; Jongmin Woo, University of North Carolina at Greensboro, Kannapolis, NC, United States; Joshua N. Adkins, Pacific Northwest National Laboratory, Richland, WA, United States; Thomas J. Mariani, University of Rochester School of Medicine and Dentistry, Rochester, NY, United States; Geremy C. Clair, Pacific Northwest National Laboratory, Richland, WA, United States; Gloria Pryhuber, University of Rochester School of Medicine and Dentistry, Rochester, NY, United States
Associate Professor University of Rochester University of Rochester Rochester, New York, United States
Background: Bronchopulmonary Dysplasia (BPD) is a disease of prematurity related to the arrest of normal lung development. Whole lung proteomics was performed to identify individual proteins and pathways differentially regulated in BPD after the birth hospitalization. Objective: To perform whole lung proteomics and identify individual proteins and pathways differentially regulated in BPD after the birth hospitalization. Design/Methods: Pediatric human donors aged 1.3 to 3 years were classified based on history of prematurity and histopathologic severity of BPD (N=3 “Recovered” BPD [rBPD] and N=3 “Established” BPD [eBPD]) with respective full term (N=6) age-matched controls. Lung tissues were biopsied (proteomics) and/or sectioned for imaging analyses. Proteins were identified by tandem mass spectroscopy requiring >2 unique peptide fragments for confirmation and validated using Western Blot. Highly multiplexed (18 antibody panel) immunofluorescence (MxIF) microscopy was performed on lung sections to enumerate cell types and validate proteomics results. Protein abundances and MxIF cell frequencies in the different groups were compared using ANOVA (p< 0.01 significant). Cell type and ontology enrichment was performed using an in-house developed enrichment tool and/or EnrichR. Results: Proteomics detected 5746 unique proteins, 186 upregulated and 534 downregulated in eBPD vs. Control, with less proteins differentially expressed in rBPD. Cell type enrichment analyses (EnrichRcell type enrichment using the HubMAP Anatomical Structures, Cell Types, plus Biomarkers tables) suggested a loss of alveolar Type I (decreased SCEL, AQP4), alveolar Type II (decreased ABCA3, SFTPC), endothelial/capillary (decreased PECAM1, FCN3, FLT1, ACE, ERG, EDNRB), and lymphatics (decreased ADGRG3), and increase in smooth muscle (TAGLN, LDB3, PODN, ISLR) and fibroblasts (MFAP5) that were confirmed by MxIF. Immunohistochemistry also supported protein GO pathway analyses predictions of upregulated ferroptosis and coagulation in eBPD vs. Control. Finally, several extracellular matrix proteins were different (Col8A1, Col14A1, Col15A1 Col18A1) in eBPD vs. Controls. Comparative MxIF analysis demonstrates variability in BPD.
Conclusion(s): There are several dysregulated proteins and pathways consistent with abnormal lung development in eBPD. rBPD has simplified lung parenchyma but fewer differentially expressed proteins. Highly multiplexed molecular analysis of human tissue provides critical contributions to elucidation of the factors after birth that define the trajectories of BPD that are important to preventing and treating their underlying causes.
