Gene Expression Profiling Identifies Cell Proliferation and Inflammation as the Predominant Pathways Regulated by Aryl Hydrocarbon Receptor in Primary Human Fetal Lung Cells Exposed to Hyperoxia.
Exposure to hyperoxia contributes to the development of bronchopulmonary dysplasia (BPD) in premature infants. We observed that aryl hydrocarbon receptor (AhR) signaling protects newborn mice and primary fetal human pulmonary microvascular endothelial cells (HPMECs) against hyperoxic injury. Additionally, a recent genome-wide transcriptome study in a newborn mouse model of BPD identified AhR as a key regulator of hyperoxia-induced gene dysregulation. Whether the AhR similarly deregulates genes in HPMEC is unknown. Therefore, the objective of this study was to characterize transcriptome level gene expression profile in AhR-sufficient and -deficient HPMEC exposed to normoxic and hyperoxic conditions. Global gene expression profiling was performed using Illumina microarray platform and selected genes were validated by real-time RT-PCR. AhR gene expression and hyperoxia independently affected the expression of 540 and 593 genes, respectively. Two-way ANOVA further identified 85 genes that were affected by an interaction between AhR expression and exposure to hyperoxia. Kyoto Encyclopedia of Genes and Genomes (KEGG), Gene Ontology, and Reactome pathway analysis identified cell proliferation, immune function, cytokine signaling, and organ development as the major pathways affected in AhR-deficient cells. The biological processes that were significantly enriched by hyperoxia included metabolic process, stress response, signal transduction, cell cycle, and immune regulation. Cell cycle was the predominant pathway affected by the combined effect of AhR knockdown and hyperoxia. Functional analysis of cell cycle showed that AhR-deficient cells had decreased proliferation compared with AhR-sufficient cells. These findings suggest that AhR modulates hyperoxic lung injury by regulating the genes that are necessary for cell proliferation and inflammation.
*Department of Pediatrics, Section of Neonatal-Perinatal Medicine; *Department of Pediatrics, Section of Neonatal-Perinatal Medicine; shivanna@bcm.edu.
Binoy
B
Shivanna
Department of Molecular and Cell Biology.
Suman
S
Maity
*Department of Pediatrics, Section of Neonatal-Perinatal Medicine;
Shaojie
S
Zhang
*Department of Pediatrics, Section of Neonatal-Perinatal Medicine;
Ananddeep
A
Patel
*Department of Pediatrics, Section of Neonatal-Perinatal Medicine;
Weiwu
W
Jiang
*Department of Pediatrics, Section of Neonatal-Perinatal Medicine;
Lihua
L
Wang
*Department of Pediatrics, Section of Neonatal-Perinatal Medicine;
Stephen E
SE
Welty
Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, Texas 77030.
John
J
Belmont
Department of Molecular and Cell Biology.
Cristian
C
Coarfa
*Department of Pediatrics, Section of Neonatal-Perinatal Medicine;
Bhagavatula
B
Moorthy
D051792:Basic Helix-Loop-Helix Proteins / Q000235:genetics* / Q000378:metabolism; D001997:Bronchopulmonary Dysplasia / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D049109:Cell Proliferation; D002478:Cells, Cultured; D019295:Computational Biology; D030541:Databases, Genetic; D020869:Gene Expression Profiling / Q000379:methods; D005786:Gene Expression Regulation; D053263:Gene Regulatory Networks; D006801:Humans; D018496:Hyperoxia / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D008168:Lung / Q000378:metabolism* / Q000473:pathology; D020411:Oligonucleotide Array Sequence Analysis; D011014:Pneumonia / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D034622:RNA Interference; D060888:Real-Time Polymerase Chain Reaction; D018336:Receptors, Aryl Hydrocarbon / Q000235:genetics* / Q000378:metabolism; D015398:Signal Transduction; D013997:Time Factors; D059467:Transcriptome; D014162:Transfection
D051792:Basic Helix-Loop-Helix Proteins; D018336:Receptors, Aryl Hydrocarbon; C111235:AHR protein, human
aryl hydrocarbon receptor; fetal human lung cells; gene expression profile; hyperoxia; organ development.