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- W3214826373 abstract "Introduction: The ductus arteriosus (DA) connects the fetal pulmonary artery and aorta, diverting placentally oxygenated blood from the developing lungs to the systemic circulation. The DA constricts in response to increased oxygen (O 2 ) immediately post-partum, resulting in functional DA closure, with anatomic closure ensuing within the first days of life. Failure of DA closure results in patent ductus arteriosus (PDA), a common complication of extreme preterm birth. The DA response to O 2 , though modulated by the endothelium, is intrinsic to DA smooth muscle cells (DASMC). DA constriction is mediated by mitochondria-derived reactive oxygen species, which increase in proportion to PaO 2 . The molecular signature of the DASMC response to O 2 is not well understood. Methods & Results: DASMC were isolated from DA obtained from 10 infants at the time of congenital heart surgery. Cells were purified by flow cytometry, negatively sorted to eliminate fibroblasts (CD90+) or endothelial cells (CD31+). DASMC purity was confirmed by α-smooth muscle actin staining. Cells were grown for 96 hours in hypoxia (2.5% O 2 ) or normoxia (19% O 2 ) and confocal calcium imaging confirmed O 2 -responsiveness of all cell lines: O 2 exposure increased intracellular calcium 18.1%±4.4% and decreased cell length 27%±1.5% relative to hypoxic baseline. RNA sequencing (Seq) of the cells grown in hypoxia and normoxia revealed significant regulation of 1344 genes (corrected p<0.05). Gene Ontology (GO) analysis provided an unbiased assessment of genes differentially regulated by O 2 , revealing enrichment of functional groups including mitochondria, cellular respiration and transcription. Multiple subunits of electron transport chain complex I, NDUF (NADH:ubiquinone oxidoreductase) family genes, are upregulated in hDASMC following normoxia. Upregulated NDUF genes were validated with qPCR (t-test p<0.05): NDUFS5 (Seq = 0.62-fold, PCR = 3.8-fold), NDUFA9 (Seq = 0.8-fold, Seq = 4.3-fold), NDUFA13 (Seq = 0.7-fold, PCR = 6.1-fold), and NDUFS8 (Seq = 0.6-fold, PCR = 7.1-fold). Conclusion: This first examination of the effects of O 2 on human DA transcriptomics supports a putative role for mitochondria as O 2 sensors, with mitochondrial subunits as potential future therapeutic targets for PDA." @default.
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- W3214826373 date "2021-11-16" @default.
- W3214826373 modified "2023-09-27" @default.
- W3214826373 title "Abstract 12927: The Human Ductus Arteriosus Smooth Muscle Cell Transcriptome Indicates a Central Role for Mitochondria in Postnatal Oxygen Sensing" @default.
- W3214826373 doi "https://doi.org/10.1161/circ.144.suppl_1.12927" @default.
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