Orphan Nuclear Receptor Family 4A (NR4A) Members NR4A2 and NR4A3 Selectively Modulate Elements of the Monocyte Response to Buffered Hypercapnia

Hypercapnia occurs when the partial pressure of carbon dioxide (CO₂) in the blood exceeds 45 mmHg. Hypercapnia is associated with several lung pathologies and is transcriptionally linked to suppression of immune and inflammatory signalling through poorly understood mechanisms. Here we propose Orphan Nuclear Receptor Family 4A (NR4A) family members NR4A2 and NR4A3 as potential transcriptional regulators of the cellular response to hypercapnia in monocytes. Using a THP-1 monocyte model, we investigated the sensitivity of NR4A family members to CO₂ and the impact of depleting NR4A2 and NR4A3 on the monocyte response to buffered hypercapnia (10% CO₂) using RNA-sequencing. We observed that NR4A2 and NR4A3 are CO₂-sensitive transcription factors and that depletion of NR4A2 and NR4A3 led to reduced CO₂-sensitivity of mitochondrial and heat shock protein (Hsp)-related genes, respectively. Several CO₂-sensitive genes were, however, refractory to depletion of NR4A2 and NR4A3, indicating that NR4As regulate certain elements of the cellular response to buffered hypercapnia but that other transcription factors also contribute. Bioinformatic analysis of conserved CO₂-sensitive genes implicated several novel putative CO₂-sensitive transcription factors, of which the ETS Proto-Oncogene 1 Transcription Factor (ETS-1) was validated to show increased nuclear expression in buffered hypercapnia. These data give significant insights into the understanding of immune responses in patients experiencing hypercapnia.