title: Acute Respiratory Distress Syndrome Neutrophils Have a Distinct Phenotype and Are Resistant to Phosphoinositide 3-Kinase Inhibition.
abstract: Acute respiratory distress syndrome is refractory to pharmacological intervention. Inappropriate activation of alveolar neutrophils is believed to underpin this disease's complex pathophysiology, yet these cells have been little studied.
  
  
  
  To examine the functional and transcriptional profiles of patient blood and alveolar neutrophils compared with healthy volunteer cells, and to define their sensitivity to phosphoinositide 3-kinase inhibition.
  
  
  
  Twenty-three ventilated patients underwent bronchoalveolar lavage. Alveolar and blood neutrophil apoptosis, phagocytosis, and adhesion molecules were quantified by flow cytometry, and oxidase responses were quantified by chemiluminescence. Cytokine and transcriptional profiling were used in multiplex and GeneChip arrays.
  
  
  
  Patient blood and alveolar neutrophils were distinct from healthy circulating cells, with increased CD11b and reduced CD62L expression, delayed constitutive apoptosis, and primed oxidase responses. Incubating control cells with disease bronchoalveolar lavage recapitulated the aberrant functional phenotype, and this could be reversed by phosphoinositide 3-kinase inhibitors. In contrast, the prosurvival phenotype of patient cells was resistant to phosphoinositide 3-kinase inhibition. RNA transcriptomic analysis revealed modified immune, cytoskeletal, and cell death pathways in patient cells, aligning closely to sepsis and burns datasets but not to phosphoinositide 3-kinase signatures.
  
  
  
  Acute respiratory distress syndrome blood and alveolar neutrophils display a distinct primed prosurvival profile and transcriptional signature. The enhanced respiratory burst was phosphoinositide 3-kinase-dependent but delayed apoptosis and the altered transcriptional profile were not. These unexpected findings cast doubt over the utility of phosphoinositide 3-kinase inhibition in acute respiratory distress syndrome and highlight the importance of evaluating novel therapeutic strategies in patient-derived cells.
authors.affiliation: 1 Department of Medicine, University of Cambridge, Cambridge, United Kingdom.
authors.forename: Jatinder K
authors.initials: JK
authors.lastname: Juss
authors.affiliation: 2 Refractory Respiratory Inflammation Discovery Performance Unit and.
authors.forename: David
authors.initials: D
authors.lastname: House
authors.affiliation: 2 Refractory Respiratory Inflammation Discovery Performance Unit and.
authors.forename: Augustin
authors.initials: A
authors.lastname: Amour
authors.affiliation: 2 Refractory Respiratory Inflammation Discovery Performance Unit and.
authors.forename: Malcolm
authors.initials: M
authors.lastname: Begg
authors.affiliation: 3 Cambridge University Hospitals NHS Foundation Trust, Cambridge, United Kingdom.
authors.forename: Jurgen
authors.initials: J
authors.lastname: Herre
authors.affiliation: 1 Department of Medicine, University of Cambridge, Cambridge, United Kingdom.
authors.forename: Daniel M L
authors.initials: DM
authors.lastname: Storisteanu
authors.affiliation: 1 Department of Medicine, University of Cambridge, Cambridge, United Kingdom.
authors.forename: Kim
authors.initials: K
authors.lastname: Hoenderdos
authors.affiliation: 4 Target Sciences, GlaxoSmithKline, Stevenage, United Kingdom; and.
authors.forename: Glyn
authors.initials: G
authors.lastname: Bradley
authors.affiliation: 4 Target Sciences, GlaxoSmithKline, Stevenage, United Kingdom; and.
authors.forename: Mark
authors.initials: M
authors.lastname: Lennon
authors.affiliation: 1 Department of Medicine, University of Cambridge, Cambridge, United Kingdom.
authors.forename: Charlotte
authors.initials: C
authors.lastname: Summers
authors.affiliation: 2 Refractory Respiratory Inflammation Discovery Performance Unit and.
authors.forename: Edith M
authors.initials: EM
authors.lastname: Hessel
authors.affiliation: 1 Department of Medicine, University of Cambridge, Cambridge, United Kingdom.
authors.forename: Alison
authors.initials: A
authors.lastname: Condliffe
authors.affiliation: 1 Department of Medicine, University of Cambridge, Cambridge, United Kingdom.
authors.forename: Edwin R
authors.initials: ER
authors.lastname: Chilvers
mesh_terms: D001992:Bronchoalveolar Lavage Fluid / Q000166:cytology; D039481:CD11b Antigen / Q000378:metabolism; D016207:Cytokines / Q000378:metabolism; D005260:Female; D005434:Flow Cytometry; D020869:Gene Expression Profiling; D006801:Humans; D019041:L-Selectin / Q000378:metabolism; D008297:Male; D008875:Middle Aged; D009504:Neutrophils / Q000187:drug effects* / Q000378:metabolism / Q000502:physiology; D020411:Oligonucleotide Array Sequence Analysis; D010641:Phenotype; D000081082:Phosphoinositide-3 Kinase Inhibitors; D012128:Respiratory Distress Syndrome / Q000188:drug therapy / Q000276:immunology*
chemical_list: D039481:CD11b Antigen; D016207:Cytokines; C509295:ITGAM protein, human; D000081082:Phosphoinositide-3 Kinase Inhibitors; D019041:L-Selectin
keywords: acute respiratory distress syndrome; neutrophil function; phosphoinositide 3-kinase