title: Identification of benzopyrone as a common structural feature in compounds with anti-inflammatory activity in a zebrafish phenotypic screen.
abstract: Neutrophils are essential for host defence and are recruited to sites of inflammation in response to tissue injury or infection. For inflammation to resolve, these cells must be cleared efficiently and in a controlled manner, either by apoptosis or reverse migration. If the inflammatory response is not well-regulated, persistent neutrophils can cause damage to host tissues and contribute to the pathogenesis of chronic inflammatory diseases, which respond poorly to current treatments. It is therefore important to develop drug discovery strategies that can identify new therapeutics specifically targeting neutrophils, either by promoting their clearance or by preventing their recruitment. Our recent in vivo chemical genetic screen for accelerators of inflammation resolution identified a subset of compounds sharing a common chemical signature, the bicyclic benzopyrone rings. Here, we further investigate the mechanisms of action of the most active of this chemical series, isopimpinellin, in our zebrafish model of neutrophilic inflammation. We found that this compound targets both the recruitment and resolution phases of the inflammatory response. Neutrophil migration towards a site of injury is reduced by isopimpinellin and this occurs as a result of PI3K inhibition. We also show that isopimpinellin induces neutrophil apoptosis to drive inflammation resolution in vivo using a new zebrafish reporter line detecting in vivo neutrophil caspase-3 activity and allowing quantification of flux through the apoptotic pathway in real time. Finally, our studies reveal that clinically available 'cromones' are structurally related to isopimpinellin and have previously undescribed pro-resolution activity in vivo These findings could have implications for the therapeutic use of benzopyrones in inflammatory disease.
authors.affiliation: The Bateson Centre, University of Sheffield, Sheffield, S10 2TN, UK Department of Infection, Immunity and Cardiovascular Disease, University of Sheffield, Sheffield, S10 2TN, UK Stem Cell Program and Division of Hematology/Oncology, Children's Hospital Boston, Howard Hughes Medical Institute, Harvard Stem Cell Institute, Harvard Medical School, Boston, 02115 MA, USA.
authors.forename: Anne L
authors.initials: AL
authors.lastname: Robertson
authors.affiliation: The Bateson Centre, University of Sheffield, Sheffield, S10 2TN, UK Department of Infection, Immunity and Cardiovascular Disease, University of Sheffield, Sheffield, S10 2TN, UK.
authors.forename: Nikolay V
authors.initials: NV
authors.lastname: Ogryzko
authors.affiliation: The Bateson Centre, University of Sheffield, Sheffield, S10 2TN, UK Department of Infection, Immunity and Cardiovascular Disease, University of Sheffield, Sheffield, S10 2TN, UK.
authors.forename: Katherine M
authors.initials: KM
authors.lastname: Henry
authors.affiliation: The Bateson Centre, University of Sheffield, Sheffield, S10 2TN, UK Department of Infection, Immunity and Cardiovascular Disease, University of Sheffield, Sheffield, S10 2TN, UK.
authors.forename: Catherine A
authors.initials: CA
authors.lastname: Loynes
authors.affiliation: Department of Infection, Immunity and Cardiovascular Disease, University of Sheffield, Sheffield, S10 2TN, UK Department of Chemistry, University of Sheffield, Sheffield, S10 2TN, UK.
authors.forename: Matthew J
authors.initials: MJ
authors.lastname: Foulkes
authors.affiliation: Department of Chemistry, University of Sheffield, Sheffield, S10 2TN, UK.
authors.forename: Marco M
authors.initials: MM
authors.lastname: Meloni
authors.affiliation: Wishtech Medical Technology, Weihai, Shandong, 264200, China.
authors.forename: Xingang
authors.initials: X
authors.lastname: Wang
authors.affiliation: Department of Musculoskeletal Biology, Institute of Ageing and Chronic Disease, University of Liverpool, Liverpool, L7 9TX, UK MRC Arthritis Research UK Centre for Integrated Research into Musculoskeletal Ageing, University of Liverpool, Liverpool, L7 9TX, UK.
authors.forename: Christopher
authors.initials: C
authors.lastname: Ford
authors.affiliation: Department of Musculoskeletal Biology, Institute of Ageing and Chronic Disease, University of Liverpool, Liverpool, L7 9TX, UK MRC Arthritis Research UK Centre for Integrated Research into Musculoskeletal Ageing, University of Liverpool, Liverpool, L7 9TX, UK.
authors.forename: Malcolm
authors.initials: M
authors.lastname: Jackson
authors.affiliation: Institute of Molecular and Cell Biology, 61 Biopolis Drive, Proteos, Singapore, 138673, Singapore.
authors.forename: Philip W
authors.initials: PW
authors.lastname: Ingham
authors.affiliation: The Bateson Centre, University of Sheffield, Sheffield, S10 2TN, UK Department of Infection, Immunity and Cardiovascular Disease, University of Sheffield, Sheffield, S10 2TN, UK.
authors.forename: Heather L
authors.initials: HL
authors.lastname: Wilson
authors.affiliation: Institute of Human Development, University of Manchester, Manchester, M13 9PL, UK.
authors.forename: Stuart N
authors.initials: SN
authors.lastname: Farrow
authors.affiliation: Faculty of Medicine, National Heart and Lung Institute, Imperial College London, Norfolk Place, London, W2 1NY, UK.
authors.forename: Roberto
authors.initials: R
authors.lastname: Solari
authors.affiliation: William Harvey Research Institute, Queen Mary University of London, Charterhouse Square, London, EC1M 6BQ, UK.
authors.forename: Roderick J
authors.initials: RJ
authors.lastname: Flower
authors.affiliation: Department of Chemistry, University of Sheffield, Sheffield, S10 2TN, UK.
authors.forename: Simon
authors.initials: S
authors.lastname: Jones
authors.affiliation: MRC/UoE Centre for Inflammation Research, University of Edinburgh, The Queen's Medical Research Institute, Edinburgh, EH16 4TJ, UK.
authors.forename: Moira K B
authors.initials: MK
authors.lastname: Whyte
authors.affiliation: The Bateson Centre, University of Sheffield, Sheffield, S10 2TN, UK Department of Infection, Immunity and Cardiovascular Disease, University of Sheffield, Sheffield, S10 2TN, UK MRC Arthritis Research UK Centre for Integrated Research into Musculoskeletal Ageing, University of Liverpool, Liverpool, L7 9TX, UK s.a.renshaw@sheffield.ac.uk.
authors.forename: Stephen A
authors.identifier: 0000-0003-1790-1641
authors.initials: SA
authors.lastname: Renshaw
mesh_terms: D000818:Animals; D000893:Anti-Inflammatory Agents / Q000737:chemistry* / Q000494:pharmacology*; D000975:Antioxidants / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects; D003374:Coumarins / Q000737:chemistry* / Q000494:pharmacology*; D004205:Cromolyn Sodium / Q000737:chemistry / Q000494:pharmacology; D004353:Drug Evaluation, Preclinical; D011564:Furocoumarins / Q000737:chemistry / Q000494:pharmacology; D007249:Inflammation / Q000473:pathology; D020556:Neutrophil Infiltration / Q000187:drug effects; D009504:Neutrophils / Q000166:cytology / Q000187:drug effects; D010641:Phenotype; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D013329:Structure-Activity Relationship; D015027:Zebrafish / Q000378:metabolism*
chemical_list: D000893:Anti-Inflammatory Agents; D000975:Antioxidants; D003374:Coumarins; D011564:Furocoumarins; C015304:isopimpinellin; D019869:Phosphatidylinositol 3-Kinases; D004205:Cromolyn Sodium
keywords: Benzopyrone; Chromone; Inflammation; Neutrophil apoptosis; Zebrafish