RNA Sequencing Analysis of Monocytes Exposed to Airway Fluid From Children With Pediatric Acute Respiratory Distress Syndrome

• Type of Material: Article
• Authors: Jocelyn R. Grunwell, Emory University; Min Huang, Emory University; Susan T. Stephenson, Emory University; Mallory Tidwell, Children's Healthcare of Atlanta; Michael J. Ripple, Emory University; Anne M Fitzpatrick, Emory University; Rishikesan Kamaleswaran, Emory University
• Language: English
• Date: 2024-10-04
• Publisher: Wolters Kluwer Health, Inc.
• Publication Version: Final Published Version
• Copyright Statement: © 2024 The Authors.
• License: Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International
• Final Published Version (URL): https://doi.org/10.1097/CCE.0000000000001125
• Title of Journal or Parent Work: Critical Care Explorations
• Volume: 6
• Issue: 10
• Start Page: e1125
• Grant/Funding Agency: National Institutes of Health; Emory University
• Grant/Funding Information: This study was supported in part by the Emory Integrated Genomics Core, which is subsidized by the Emory University School of Medicine and is one of the Emory Integrated Core Facilities. Additional support was provided by the Georgia Clinical & Translational Science Alliance of the National Institutes of Health (NIH) under Award Number UL1TR002378. NIH provided funding grants K23 HL151897 to Dr. Grunwell. Funding was provided by the NIH grant K24 NR018866 to Dr. Fitzpatrick.
• Keywords: transcriptomics; tracheal aspirate; RNA sequencing; pediatric acute respiratory distress syndrome; pediatric intensive care; monocytes
• Subject - Topics: Pediatrics; Genetics

Abstract

OBJECTIVES: Monocytes are plastic cells that assume different polarization states that can either promote inflammation or tissue repair and inflammation resolution. Polarized monocytes are partially defined by their transcriptional profiles that are influenced by environmental stimuli. The airway monocyte response in pediatric acute respiratory distress syndrome (PARDS) is undefined. To identify differentially expressed genes and networks using a novel transcriptomic reporter assay with donor monocytes exposed to the airway fluid of intubated children with and at-risk for PARDS. To determine differences in gene expression at two time points using the donor monocyte assay exposed to airway fluid from intubated children with PARDS obtained 48–96 hours following initial tracheal aspirate sampling. DESIGN: In vitro pilot study carried out using airway fluid supernatant. SETTING: Academic 40-bed PICU. PARTICIPANTS: Fifty-seven children: 44 children with PARDS and 13 children at-risk for PARDS. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: We performed bulk RNA sequencing using a transcriptomic reporter assay of monocytes exposed to airway fluid from intubated children to discover gene networks differentiating PARDS from at-risk for PARDS and those differentiating mild/moderate from severe PARDS. We also report differences in gene expression in children with PARDS 48–96 hours following initial tracheal aspirate sampling. We found that interleukin (IL)-10, IL-4, and IL-13, cytokine/chemokine signaling, and the senescence-associated secretory phenotype are upregulated in monocytes exposed to airway fluid from intubated children with PARDS compared with those at-risk for PARDS. Signaling by NOTCH, histone deacetylation/acetylation, DNA methylation, chromatin modifications (B-WICH complex), and RNA polymerase I transcription and its associated regulatory apparatus were upregulated in children with PARDS 48–96 hours following initial tracheal aspirate sampling. CONCLUSIONS: We identified gene networks important to the PARDS airway immune response using bulk RNA sequencing from a monocyte reporter assay that exposed monocytes to airway fluid from intubated children with and at-risk for PARDS. Mechanistic investigations are needed to validate our findings.