The circular RNA landscape in specific peripheral blood mononuclear cells of critically ill patients with sepsis
Hina N. Khan,
Xanthe Brands,
Simona Aufiero,
Arie J. Hoogendijk,
Augustijn M. Klarenbeek,
Tjitske S. R. van Engelen,
Bastiaan W. Haak,
Lonneke A. van Vught,
Janneke Horn,
Marcus J. Schultz,
Aeilko H. Zwinderman,
Tom van der Poll,
Brendon P. Scicluna
Affiliations
Hina N. Khan
Center for Experimental Molecular Medicine, Amsterdam University Medical Centers, location Academic Medical Center, University of Amsterdam
Xanthe Brands
Center for Experimental Molecular Medicine, Amsterdam University Medical Centers, location Academic Medical Center, University of Amsterdam
Simona Aufiero
Department of Clinical Epidemiology, Biostatistics and Bioinformatics, Amsterdam University Medical Centers, location Academic Medical Center, University of Amsterdam
Arie J. Hoogendijk
Center for Experimental Molecular Medicine, Amsterdam University Medical Centers, location Academic Medical Center, University of Amsterdam
Augustijn M. Klarenbeek
Center for Experimental Molecular Medicine, Amsterdam University Medical Centers, location Academic Medical Center, University of Amsterdam
Tjitske S. R. van Engelen
Center for Experimental Molecular Medicine, Amsterdam University Medical Centers, location Academic Medical Center, University of Amsterdam
Bastiaan W. Haak
Center for Experimental Molecular Medicine, Amsterdam University Medical Centers, location Academic Medical Center, University of Amsterdam
Lonneke A. van Vught
Center for Experimental Molecular Medicine, Amsterdam University Medical Centers, location Academic Medical Center, University of Amsterdam
Janneke Horn
Department of Intensive Care Medicine, Amsterdam University Medical Centers, location Academic Medical Center, University of Amsterdam
Marcus J. Schultz
Department of Intensive Care Medicine, Amsterdam University Medical Centers, location Academic Medical Center, University of Amsterdam
Aeilko H. Zwinderman
Department of Clinical Epidemiology, Biostatistics and Bioinformatics, Amsterdam University Medical Centers, location Academic Medical Center, University of Amsterdam
Tom van der Poll
Center for Experimental Molecular Medicine, Amsterdam University Medical Centers, location Academic Medical Center, University of Amsterdam
Brendon P. Scicluna
Center for Experimental Molecular Medicine, Amsterdam University Medical Centers, location Academic Medical Center, University of Amsterdam
Abstract Background Dysregulation of the host immune response is a pathognomonic feature of sepsis. Abnormal physiological conditions are understood to shift efficient linear splicing of protein-coding RNA towards non-canonical splicing, characterized by the accumulation of non-coding circularized (circ)RNA. CircRNAs remain unexplored in specific peripheral blood mononuclear cells (PBMCs) during sepsis. We here sought to identify and characterize circRNA expression in specific PBMCs of patients with sepsis due to community-acquired pneumonia (CAP) relative to healthy subjects. Methods The study comprised a discovery cohort of six critically ill patients diagnosed with sepsis due to community-acquired pneumonia and four (age, gender matched) healthy subjects. PBMCs were isolated, and fluorescence-activated cell sorting was used to purify CD14+ monocytes, CD4+, CD8+ T cells, and CD19+ B cells for RNA sequencing. CD14+ monocytes from independent six healthy volunteers were purified, and total RNA was treated with or without RNase R. Results RNA sequencing of sorted CD14+ monocytes, CD4+, CD8+ T cells, and CD19+ B cells from CAP patients and healthy subjects identified various circRNAs with predominantly cell-specific expression patterns. CircRNAs were expressed to a larger extent in monocytes than in CD4+, CD8+ T cells, or B cells. Cells from CAP patients produced significantly higher levels of circRNA as compared to healthy subjects. Considering adjusted p values, circVCAN (chr5:83519349-83522309) and circCHD2 (chr15:93000512-93014909) levels in monocytes were significantly altered in sepsis. Functional inference per cell-type uncovered pathways mainly attuned to cell proliferation and cytokine production. In addition, our data does not support a role for these circRNAs in microRNA sequestration. Quantitative PCR analysis in purified monocytes from an independent group of healthy volunteers confirmed the existence of circVCAN and circCHD2. Conclusions We provide a benchmark map of circRNA expression dynamics in specific immune cell subsets of sepsis patients secondary to CAP. CircRNAs were more abundant in immune cells of sepsis patients relative to healthy subjects. Further studies evaluating circRNA expression in larger cohorts of sepsis patients are warranted.
