Journal Article

Glucocorticoid Receptor ChIP-Seq Identifies PLCD1 as a KLF15 Target that Represses Airway Smooth Muscle Hypertrophy

Sasse, Sarah K., Panettieri, Reynold A., Phang, Tzu L., Kadiyala, Vineela, Danhorn, Thomas, Gerber, Anthony N.

2017

Overview

Glucocorticoids exert important therapeutic effects on airway smooth muscle (ASM), yet few direct targets of glucocorticoid signaling in ASM have been definitively identified. Here, we show that the transcription factor, Krüppel-like factor 15 (KLF15), is directly induced by glucocorticoids in primary human ASM, and that KLF15 represses ASM hypertrophy. We integrated transcriptome data from KLF15 overexpression with genome-wide analysis of RNA polymerase (RNAP) II and glucocorticoid receptor (GR) occupancy to identify phospholipase C delta 1 as both a KLF15-regulated gene and a novel repressor of ASM hypertrophy. Our chromatin immunoprecipitation sequencing data also allowed us to establish numerous direct transcriptional targets of GR in ASM. Genes with inducible GR occupancy and putative antiinflammatory properties included IRS2, APPL2, RAMP1, and MFGE8. Surprisingly, we also observed GR occupancy in the absence of supplemental ligand, including robust GR binding peaks within the IL11 and LIF loci. Detection of antibody-GR complexes at these areas was abrogated by dexamethasone treatment in association with reduced RNA polymerase II occupancy, suggesting that noncanonical pathways contribute to cytokine repression by glucocorticoids in ASM. Through defining GR interactions with chromatin on a genome-wide basis in ASM, our data also provide an important resource for future studies of GR in this therapeutically relevant cell type.

• Publisher: American Thoracic Society
• Subject: Adenoviridae - genetics / Airway management / Airway Remodeling - genetics / Asthma / Binding sites / Cells, Cultured / Chromatin / Chromatin Immunoprecipitation / Cytokines / Dexamethasone / Dexamethasone - pharmacology / DNA-directed RNA polymerase / Gene expression / Gene Expression Regulation - drug effects / Gene Expression Regulation - physiology / Genes, Reporter / Genomes / Glucocorticoids / Humans / Hypertrophy / Immunoprecipitation / Interleukin 1 / Interleukin 11 / Kruppel-Like Transcription Factors - physiology / Ligands / Metabolism / Muscle, Smooth - metabolism / Muscle, Smooth - pathology / Musculoskeletal system / Nuclear Proteins - physiology / Original Research / Phospholipase C / Phospholipase C delta - genetics / Phospholipase C delta - physiology / Plasmids / Primary Cell Culture / Receptors, Glucocorticoid - physiology / Recombinant Fusion Proteins - metabolism / Respiratory System - cytology / Respiratory tract / RNA polymerase / RNA Polymerase II - metabolism / Rodents / Sequence Analysis, RNA / Smooth muscle / Transcriptome / Transduction, Genetic / Transforming Growth Factor beta - pharmacology