Professor Edwin Chilvers

Research

The aims of my current research are to determine the molecular mechanisms underlying neutrophil priming, the functional role of the gamma and delta isoforms of phosphoinositide 3OH-kinase in myeloid cells, and the ability of hypoxia and inflammatory cytokines to inhibit neutrophil apoptosis. These studies aim to provide new insights into the mechanisms of inflammatory disease in the lung and other organs and information about the cellular and molecular processes regulating neutrophil priming, de-priming and apoptosis.

Neutrophil priming is an event whereby the response of the cell to an activating stimulus is augmented greatly by prior exposure to a priming agent (eg. LPS, TNF-alpha) and is a critical determinant of the pathogenic capacity of these cells in vivo. We have demonstrated a cardinal role for phosphoinositide 3-kinase (PI3-kinase) activation and PtdIns(3,4,5)P3 generation in triggering superoxide anion release in neutrophils and that up-regulation of this pathway is a central mechanism underlying priming.

In collaboration with Drs Hawkins and Stephens at the Babraham Institute, we have shown that the myeloid restricted Gbeta/gamma-regulated PI3-kinase (p101/PI3-kinase-gamma) is the principal source of agonist-stimulated PtdIns(3,4,5)P3 in neutrophils, work that involved the generation of a unique p101/ PI3-kinase-gamma mouse knockout. These studies also revealed the mechanism whereby PtdIns(3,4,5)P3 activates the NADPH oxidase namely via a direct interaction between PtdIns(3)P and the PX domain of the p40phox oxidase subunit. Our lab also revealed for the first time that neutrophil priming is reversible and that these cells can participate in a complete cycle of priming, de-priming and re-priming. This observation refuted the view that priming is a terminal event and suggested that neutrophils primed in vivo can be retained in the pulmonary circulation and allowed to de-prime before re-entering the circulation. This has been confirmed using laser trapping of neutrophils where we have preliminary data to suggest that de-priming can be accelerated by repetitive mechanical stretch.

To test the above hypothesis we have developed techniques using 111-Indium and 99m-Technicium-labelled autologous granulocytes (both neutrophils and eosinophils), which have allowed us to make the first measurements of single pass neutrophil transit across the lung in man; these confirm major initial retention of ex-vivo primed cells by the pulmonary circulation followed by slow but complete release. These studies have now been extended into studying human eosinophil kinetics in vivo, where we hope to develop clinically useful eosinophil scanning and techniques to quantify lung-specific eosinophil uptake. We have also demonstrated that priming and hypoxia have profound effects on neutrophil longevity. These studies have shown that in both human and murine cells that Class I PI3-kinases and NF-kappaB play dominant roles in this survival effect and have provided the first demonstration of hypoxic sensing in neutrophils together with a molecular explanation for the profound survival effect of hypoxia in neutrophils. These studies predict that the myeloid enriched PI3-kinase-gamma and the hypoxia-PHD-HIF-alpha pathways represent important pharmacological targets in granulocytic inflammation. These studies also underpin our current interests in using radiolabelled autologous human cells to study granulocyte trafficking in vivo.

Publications

Key publications:

Sowerby JM, Thomas DC, Clare S, Espeli M, Guerrero JA, Hoenderdos K, Harcourt K, Marsden M, Abdul-Karim J, Clement M, Antrobus R, Umrania Y, Barton P, Juss JK, Flint S, Condliffe AM, Lyons PA, Humphreys I, Chilvers ER, Ouwehand W, Dougan G, Smith KGC. NBEAL2 is required for neutrophil and NK cell function and pathogen defence. J Clin Invest 2017; in press.

Mitrofan CG, Appleby SL, Nash GB, Mallat Z, Chilvers ER, Upton PD, Morrell NW. Bone morphogenetic protein (BMP) 9 and BMP10 enhance TNFα-induced monocyte recruitment to the vascular endothelium predominantly via activin receptor-like kinase 2. J Biol Chem 2017; in press. pii: jbc.M117.778506. doi: 10.1074/jbc.M117.778506.

Walmsley SR, Thompson AA, Marriott HM, Shaw G, Parmar S, Sabroe I, Dockrell DH Taylor CT, Schneider M, Chilvers ER, Pugh CW, Ratcliffe PJ, Maxwell PH, Mazzone M, Carmeliet P, Whyte MKB. Enhancement of glycolysis and glycogen stores by myeloid-specific PHD2 inactivation leads to excessive neutrophilic responses. J Clin Invest 2017; in press.

Porter LM, Farahi N, Cowburn AS, Deighton, Juss JK, Farrow SN, Condliffe AM, Chilvers ER. Hypoxia induces cytokine secretion and a quasi-corticosteroid resistant state in human eosinophils. Clin Exp Allergy 2017; 47(6): 770-784. doi: 10.1111/cea.12877.

Koop G, Vrieling M, Storisteanu D, Lok L, Monie T, Ba I, van Wigcheren G, X, Raisen C, Gleadall N, Timmerman NH, Jaap A. Wagenaar JA, Klunder HM, Raghunathan D, Fitzgerald R, Zadoks R, Paterson GK, Torres C, Waller AS, Loeffler A, Loncaric I, Hoet AE, Bergström K, De Martino L, Pomba C, de Lencastre H, Richardson EJ, Chilvers ER, De Haas C, Van Kessel K, van Strijp JAG, Harrison EM, Holmes MA. Equid isolates of Staphylococcus aureus harbour a novel phage encoded leukocidin LukPQ, a potent and specific killer of equine neutrophils. Science Rep 2017; 7: 40660. doi: 10.1038/srep40660.

Thompson AAR, Dickinson RS, Marriott HM, Williams L, Murphy F, Lewis A, Hameed A, Preston JA, Lawrie A, Finisguerra V, Mazzone M, Simon MC, Foster SJ, Chilvers ER, Cowburn AS, Dockrell DH, Johnson RS, Whyte MKB, Walmsley SR. Hypoxia determines survival outcomes of bacterial infection through HIF-1alpha dependent re-programming of leukocyte metabolism. Science Immunol 2017; 2(8). pii: eaal2861. doi: 10.1126/sciimmunol.aal2861.

Thomas DC#, Clare S#, Sowerby JM, Pardo M, Juss JK, Goulding DA, Van der Weyden L, Storisteanu DML, Prakash A, Espeli M, Flint S, Lee JC, Kane L, Hoenderdos K, Adams D, Bateman A, Chaudhry J, Lyons PA, Condliffe AM, Chilvers ER, Dougan G, Smith KGC (#Joint first authors). Eros, a novel trans-membrane protein, controls the reactive oxygen burst and is essential for innate immunity. J Exp Med 2017; 214: 1111-1128. doi: 10.1084/jem.20161382.

Farahi N, Paige E, Balla J, Prudence E, Ferreira RC, Southwood M, Appleby S, Bakke P, Gulscik A, Litonjua G, Sparrow D, Silverman E, Cho M, Paul D, Danesh J, Freitag DF#, Chilvers ER# (# joint senior authors). Neutrophil-mediated IL-6 receptor trans-signaling and the risk of chronic obstructive pulmonary disease and related phenotypes. Hum Mol Gen 2017; 26(8): 1584-1596. doi: 10.1093/hmg/ddx053.

Ekpenyong AE, Toepfner N, Fiddler C, Herbig M, Li W, Cojoc G, Summers C, Guck J, Chilvers ER. Mechanical deformation induces depolarization of neutrophils. Science Adv 2017; 3(6): e1602536. doi: 10.1126/sciadv.1602536.

Appleby SL, Mitrofan C-G, Crosby A, Hoenderdos K, Lodge K, Upton PD, Yates CM, Nash GB, Chilvers ER#, Morrell NM# (#joint senior authors). Bone morphogenetic protein 9 enhances lipopolysaccharide-induced leukocyte recruitment to the vascular endothelium. J Immunol 2016; 197: 3304-3314. doi: 10.4049/jimmunol.1601219.

Juss JK, Herre J, Storisteanu DML, Hoenderdos K, House D, Bradley G, Amour A, Summers C, Begg M, Hesseel EM, Condliffe AM, Chilvers ER. Functional and phenotypic heterogeneity of alveolar and circulating neutrophil subsets in the acute respiratory distress syndrome. Am J Respir Crit Care Med 2016; 194: 961-973. doi: 10.1164/rccm.201509-1818OC.

Cader MZ, Kempster SL, Boroviak K, Zhang Q, Assadi G, Sewell G, Clare S, Saveljeva S, Mukhopadhyay S, Brown KP, Ashcroft JW, Tschurtschenthaler M, Raine T, Chilvers ER, Kaneider NC, Floto RA, d’Amato M, Wakelam MJ, Bradley A, Dougan G, Kaser A. C13orf31 (FAMIN) is a central regulator of immunometabolic function. Nat Immunol 2016; 17: 1046-56. doi: 10.1038/ni.3532

Cowburn AS, Crosby A, Macias D, Branco C, Colaço R, Southwood M, Crotty Alexander LE, Morrell NW, Chilvers ER, Johnson RS. Pulmonary endothelial HIF2α-arginase axis plays an essential role in the development of hypoxia pulmonary hypertension. PNAS 2016; 113(31): 8801-8806. doi: 10.1073/pnas.1602978113.

Radjabova V, Mastroeni P, Skjodt K, Zaccone P, de Bono B, Goodall C, Chilvers ER, Juss J, Jones DC, Trowsdale J, Barrow AD. TARM1 Is a Novel Leukocyte Receptor Complex-Encoded ITAM Receptor That Costimulates Proinflammatory Cytokine Secretion by Macrophages and Neutrophils. J Immunol 2015; 195: 3149-59. doi: 10.4049/jimmunol.1401847.

Cowburn AS, Crotty-Alexander LE, Nizet V, Chilvers ER, Johnson RS. Epidermal deletion of HIF-2α increases wound closure*. J Invest Dermatol 2014; 134: 801-808. doi: 10.1038/jid.2013.395.

Cowburn AS, Takeda N, Boutin AT, Kim JW, Sterling JC, Nakasaki M, Southwood M, Goldrath AW, Jamora C, Nizet V, Chilvers ER, Johnson RS.HIF isoforms in the skin differentially regulate systemic arterial pressure. Proc Natl Acad Sci U S A. 2013 Oct 22;110(43):17570-5.

Farahi N, Loutsios C, Simmonds RP, Porter L, Gillett D, Heard S, Peters AM, Condliffe AM, Chilvers ER. Measurement of eosinophil kinetics in healthy volunteers. Methods Mol Biol. 2014;1178:165-76. doi: 10.1007/978-1-4939-1016-8_15.

Guck J, Chilvers ER. Mechanics meets medicine. Sci Transl Med. 2013;5(212): 212fs41.

Thompson AA, Elks PM, Marriott HM, Eamsamarng S, Higgins KR, Lewis A, Williams L, Parmar S, Shaw G, McGrath EE, Formenti F, Van Eeden FJ, Kinnula VL, Pugh CW, Sabroe I, Dockrell DH, Chilvers ER, Robbins PA, Percy MJ, Simon MC, Johnson RS, Renshaw SA, Whyte MK, Walmsley SR. Hypoxia-inducible factor 2α regulates key neutrophil functions in humans, mice, and zebrafish.
Blood. 2014 Jan 16;123(3):366-76.

Angulo I, Vadas O, Garçon F, Banham-Hall E, Plagnol V, Leahy TR, Baxendale H, Coulter T, Curtis J, Wu C, Blake-Palmer K, Perisic O, Smyth D, Maes M, Fiddler C, Juss J, Cilliers D, Markelj G, Chandra A, Farmer G, Kielkowska A, Clark J, Kracker S, Debré M, Picard C, Pellier I, Jabado N, Morris JA, Barcenas-Morales G, Fischer A, Stephens L, Hawkins P, Barrett JC, Abinun M, Clatworthy M, Durandy A, Doffinger R, Chilvers ER, Cant AJ, Kumararatne D, Okkenhaug K, Williams RL, Condliffe A, Nejentsev S. Phosphoinositide 3-kinase δ gene mutation predisposes to respiratory infection and airway damage. Science. 2013 Nov 15;342(6160):866-71.

Herre J, Grönlund H, Brooks H, Hopkins L, Waggoner L, Murton B, Gangloff M, Opaleye O, Chilvers ER, Fitzgerald K, Gay N, Monie T, Bryant C. Allergens as immunomodulatory proteins: the cat dander protein Fel d 1 enhances TLR activation by lipid ligands. J Immunol. 2013 Aug 15;191(4):1529-35.

Farahi N, Condliffe AM, Singh NR, Heard S, Summers C, Simmonds RP, Solanki CK, Solanki K, Balan KK, Ruparelia P, Peters AM, Chilvers ER. Use of 111-Indium-labelled autologous eosinophils to establish the in vivo kinetics of human eosinophils in healthy subjects. Blood. 2012 Nov 8;120(19):4068-71

McGovern NN, Cowburn AS, Walmsley SR, Thompson R, Summers C, Willcocks LC, Whyte MKB, Condliffe AM, Chilvers ER. Hypoxia inhibits respiratory burst activity and killing of staphylococcus aureus in human neutrophils. J Immunol 2011; 186:453-63.

Walmsley SR, Chilvers ER, Thompson R Vaughan K, Marriott H, Parker L, Shaw G, Parmar S, Schneider M, Sabroe I, Dockrell D, Milo M, Taylor C, Johnson RS, Pugh CW, Ratcliffe PJ, Maxwell PH, Carmeliet P, Whyte MK. Prolyl hydroxylase PHD3 is essential for hypoxic regulation of neutrophilic inflammation. J Clin Invest 2011; 121:1053-63.

Condliffe AM, Suire SM, Ferguson JG, EllsonCD, Guillou H, Davidson K, Welch H, Coadwell J, Turner M, Chilvers^.ER, Hawkins PT, Hawkins LR. Gbgs and the Ras binding domain of p110g are both important regulators if PI(3)Kg signalling in neutrophils. Nature Cell Biol 2006; 8:1303-9.

Walmsley SR, Print C, Johnson RS, Cramer T, Sobolewski A, Condliffe A, Cowburn AS, Johnson N, Chilvers ER. Hypoxia-induced neutrophil survival is mediated by HIF-1a dependent NF-kB activity. J Exp Med 2005; 201:105-15.

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