Towards an integrated understanding of neutrophil biology and their role in resisting infection
Lead Research Organisation:
Babraham Institute
Department Name: UNLISTED
Abstract
We will have started to exploit imaging tools (eg Raichu-Rac reporter mouse strain) and have progressed our systematic gene-targetting strategy to understand spatio-temporal regulation of PI3Ks and small GTPases that is known to be essential for neutrophil migration and pathogen-killing. This activity should lead to 2 or 3 papers being either accepted or under review at the mid term review. We will have established the cell lines expressing Fc Rs and GFP-iPX construct to enable us to commence a genome-wide screen for regulators of vps34 in the context of phagosome formation in comparison to our on-going screen for regulators of vps34 upstream of autophagosome formation (see below). We will have begun to define the primary transcriptional networks of mouse neutrophils and how that is determined by different stimuli in preparation for our studies to address how some aspects of the reduced functionality of neutrophils in aged animals maybe related to changes in their transcriptional landscape. We will validate the tools to probe the identity of endogenous protein cysteine residues modified by ROS in work with other cell types and apply them to neutrophils in the later stages of the ISPG
Organisations
- Babraham Institute, United Kingdom (Lead Research Organisation)
- Academy of Sciences of the Czech Republic (Collaboration)
- University of Warwick, United Kingdom (Collaboration)
- University of California, San Francisco, United States (Collaboration)
- GlaxoSmithKline (GSK) (Collaboration)
- University of Texas at San Antonio, United States (Collaboration)
- University of Cambridge, United Kingdom (Collaboration)
- Medical Research Council (Collaboration)
- Quadram Institute Bioscience (Collaboration)
- IEEC-CSIC (Collaboration)
- Chiesi (Collaboration)
- University of Sao Paolo (Collaboration)
- Babraham Bioscience Technologies (Collaboration)
- University of Sheffield, United Kingdom (Collaboration)
- Francis Crick Institute (Collaboration)
- Akita University (Collaboration)
- Beatson Institute for Cancer Research, United Kingdom (Collaboration)
- King's College London, United Kingdom (Collaboration)
Publications
Barber MA
(2012)
The guanine-nucleotide-exchange factor P-Rex1 is activated by protein phosphatase 1a.
in The Biochemical journal
Suire S
(2012)
GPCR activation of Ras and PI3Kc in neutrophils depends on PLCb2/b3 and the RasGEF RasGRP4.
in The EMBO journal
Boyle KB
(2012)
Activation of the neutrophil NADPH oxidase by Aspergillus fumigatus.
in Annals of the New York Academy of Sciences
Hoeller O
(2013)
Two distinct functions for PI3-kinases in macropinocytosis.
in Journal of cell science
Stephens L
(2013)
More paths to PI3K?.
in PLoS biology
Gambardella L
(2013)
Phosphoinositide 3-OH kinase regulates integrin-dependent processes in neutrophils by signaling through its effector ARAP3.
in Journal of immunology (Baltimore, Md. : 1950)
Campbell AD
(2013)
P-Rex1 cooperates with PDGFRß to drive cellular migration in 3D microenvironments.
in PloS one
Angulo I
(2013)
Phosphoinositide 3-kinase d gene mutation predisposes to respiratory infection and airway damage.
in Science (New York, N.Y.)
| Description | Our work has revealed new aspects of the way the immune system works. Some of these ideas also change the way we understand how inflammatory responses arise and hence potential ways to try and reduce inflammation in disease. This includes possible pharmaceutical targets such as PI3Ks, RhoG, and PRex. Our work has underpinned recetn advances in ageing research suggesting that in neutrophils from aged people there is excessive class I PI3K activity that drives increased chemokinesis and inteferes with efficient chemotaxis. We are collaborating with the Lord and Sapey labs in Birmingham to take this forward. |
| Exploitation Route | Our work has mapped out core signalling events following activation of pro-inflammatory receptors on neutrophils. Not only does this have great value as a physiological model of GPCR signalling and immune cell activation by G-protein signalling it is also very relevant to companies attempting to reduce inflammation. Many pharmaceutical companies are attempting to generate PI3K and PRex inhibitors to treat inflammatory disease. these could lead to drugs with the potential to impact the quality of life of many patients and in some cases be life-saving therapeutics. |
| Sectors | Education,Healthcare,Pharmaceuticals and Medical Biotechnology |
| Description | Our work has led to commercial activity targetting PI3K and PRex in the immune system, including neutrophils, as potential targets for anti-inflammatories. This work led directly to collaborations with GSK including an MRC-MICA award with GSK, to understand the role of PI3Kgamma in neutrophil biology relevant to disease. |
| Sector | Education,Healthcare,Pharmaceuticals and Medical Biotechnology |
| Impact Types | Economic |
| Title | Gbg-PI3Kbeta knock-in mice |
| Description | Generation of a genetically engineered mouse strain in which mutations to the p110beta catalytic subunit of Class I PI3Kbeta were mutated to selectively block interaction with Gbg subunits of heterotrimeric G proteins. |
| Type Of Material | Model of mechanisms or symptoms - mammalian in vivo |
| Year Produced | 2015 |
| Provided To Others? | Yes |
| Impact | Houslay et al (2016) Sci Signal. 9(441):ra82. doi: 10.1126/scisignal.aae0453. |
| Description | Alison Condliffe |
| Organisation | University of Sheffield |
| Country | United Kingdom |
| Sector | Academic/University |
| PI Contribution | In vitro work with human neutrophils, measurements of phosphoinositides. |
| Collaborator Contribution | Human patient samples. |
| Impact | Publications fully reported |
| Description | Beatson Institute, Glasgow: Characterisation of our Rac-activity Fret reporter mice |
| Organisation | Beatson Institute for Cancer Research |
| Country | United Kingdom |
| Sector | Academic/University |
| PI Contribution | joint experiments exchange of reagents lab visits joint publications |
| Collaborator Contribution | joint experiments exchange of reagents lab visits joint publications |
| Impact | 2 joint papers |
| Start Year | 2011 |
| Description | Bretschneider - chemotaxis |
| Organisation | University of Warwick |
| Department | School of Life Sciences |
| Country | United Kingdom |
| Sector | Academic/University |
| PI Contribution | reagents and expertise |
| Collaborator Contribution | expertise |
| Impact | Norton et al (2016) Adv Biol Regul. 60:36-45. doi: 10.1016/j.jbior.2015.10.005 Du et al (2013) BMC Bioinformatics. 14:296. doi: 10.1186/1471-2105-14-296. Cell biology, mathematical modelling |
| Start Year | 2010 |
| Description | Chiesi |
| Organisation | Chiesi |
| Country | Italy |
| Sector | Private |
| PI Contribution | Measurements of phosphoinositides |
| Collaborator Contribution | Development of PI3K inhibitors towards clinical trials |
| Impact | Multi-disciplinary. We provide lipidomics expertise , Chiesi provide med chem and models |
| Start Year | 2016 |
| Description | Condliffe - neutrophil biology |
| Organisation | University of Cambridge |
| Department | School of Clinical Medicine |
| Country | United Kingdom |
| Sector | Academic/University |
| PI Contribution | Intellectual input, reagents, measurement of phopshoinositides |
| Collaborator Contribution | Intellectual input, personnel, reagents, clinical samples |
| Impact | Angulo et al (2013) Science. 342(6160):866-71. doi: 10.1126/science.1243292 Juss et al (2012) PLoS One. 7(9):e45933. doi: 10.1371/journal.pone.0045933 |
| Description | Curi - neutrophils and lipids |
| Organisation | University of Sao Paulo |
| Country | Brazil |
| Sector | Academic/University |
| PI Contribution | expertise, reagents, hosted exchange visits |
| Collaborator Contribution | expertise, exchange of personnel |
| Impact | Vinolo et al (2011) PLoS One. 6(6):e21205. doi: 10.1371/journal.pone.0021205. Epub 2011 Jun 15 |
| Start Year | 2008 |
| Description | Downward_RBD of Class I PI3Ks |
| Organisation | Francis Crick Institute |
| Country | United Kingdom |
| Sector | Academic/University |
| PI Contribution | We designed the study and performed all experiments |
| Collaborator Contribution | They provided genetically engineered mouse embryonic fibroblast cell lines with mutations in the Ras Binding Domain (RBD) of Class I PI3K alpha and beta. |
| Impact | Houslay et al (2016) Sci Signal. 16;9(441):ra82. doi: 10.1126/scisignal.aae0453. |
| Start Year | 2014 |
| Description | GSK/Gus |
| Organisation | GlaxoSmithKline (GSK) |
| Department | Respiratory Biology GSK |
| Country | United Kingdom |
| Sector | Private |
| PI Contribution | Research into the physiological roles of Pi3kgamma in inflammatory signalling in neutrophils |
| Collaborator Contribution | Knowledge of commercially important directions and implications of our experiments |
| Impact | In the first year of the study. |
| Start Year | 2011 |
| Description | Jonathan Clark (BBT, Cambridge): |
| Organisation | Babraham Bioscience Technologies |
| Country | United Kingdom |
| Sector | Private |
| PI Contribution | Development of a first-generation small-molecule P-Rex inhibitor |
| Collaborator Contribution | Development of a first-generation small-molecule P-Rex inhibitor |
| Impact | production of small molecule compounds to inhibit P-Rex |
| Start Year | 2009 |
| Description | KCL MSc Programme 2019/2020 |
| Organisation | King's College London |
| Country | United Kingdom |
| Sector | Academic/University |
| PI Contribution | We are hosting an MSc student from KCL Pharmacology Department for their Master's project. |
| Collaborator Contribution | Asked us to host their MSc student in the lab. |
| Impact | We have published several papers together. |
| Start Year | 2013 |
| Description | Lindsay Hall |
| Organisation | Quadram Institute Bioscience |
| Country | United Kingdom |
| Sector | Academic/University |
| PI Contribution | Discussion/advice about possible roles of inhibitory receptors on immune cells in gut |
| Collaborator Contribution | Advice about gut immune biology |
| Impact | None yet |
| Start Year | 2011 |
| Description | Orion Weiner |
| Organisation | University of California, San Francisco |
| Department | Department of Biochemistry and Biophysics |
| Country | United States |
| Sector | Academic/University |
| PI Contribution | Analysis of phosphoinositides, advice about neutrophils, reagents. |
| Collaborator Contribution | Samples, reagents, technological advice. |
| Impact | Publications fully reported. |
| Description | Role of SOS1/2 in neutrophil priming |
| Organisation | IEEC-CSIC |
| Country | Spain |
| Sector | Public |
| PI Contribution | We provided the intellectual input and performed all the experiments |
| Collaborator Contribution | They provided the SOS1/2 conditional KO mice and advice on using them |
| Impact | Publication: PMID: 30720883 |
| Start Year | 2015 |
| Description | Sasaki_ mass spec and prostate |
| Organisation | Akita University |
| Country | Japan |
| Sector | Academic/University |
| PI Contribution | Intellectual input, reagents, mass spectrometric methods, personnel |
| Collaborator Contribution | Intellectual input, reagents, mass spectrometric methods, personnel |
| Impact | Norton et al (2016) Adv Biol Regul. 60:36-45. doi: 10.1016/j.jbior.2015.10.005 Ferguson et al (2007) Nat Cell Biol. 9(1):86-91. Malek M, Kielkowska A, Chessa T, Anderson KE, Barneda D, Pir P..... Stephens LR, (2017). PTEN Regulates PI(3,4)PSignaling Downstream of Class I PI3K.. Molecular cell, 68 (3), pp. 566-580.e10 |
| Description | Sebo_cAMP Signaling of Adenylate Cyclase toxin in neutrophils |
| Organisation | Academy of Sciences of the Czech Republic |
| Department | Laboratory of Molecular Biology of Bacterial Pathogens |
| Country | Czech Republic |
| Sector | Learned Society |
| PI Contribution | We hosted a visiting student to perform some assays of neutrophil function, including the production of reactive oxygen species and the activation of some signalling pathways |
| Collaborator Contribution | They instigated the study, provided biological material and reagents and conducted experiments. |
| Impact | Cerny et al (2017) J Immunol. 198(3):1285-1296. doi: 10.4049/jimmunol.1601309. Epub 2016 Dec 30. |
| Start Year | 2013 |
| Description | Simon Pitchford |
| Organisation | King's College London |
| Department | Department of Pharmacology & Therapeutics |
| Country | United Kingdom |
| Sector | Academic/University |
| PI Contribution | Collaboration on the interactions of proinflammatory platelets with leukocytes during the recruitment of neutrophils and other types of leukocytes to sites of inflammation. Our lab has contributed the identification of platelet Rac-GEFs that are crucial for these interactions. I nthe absence of these platelet Rac-GEFs, platelet/leukocyte interactions are blocked and, in consequence, leukcoyte recruitment to sites of inflammation is also blocked. |
| Collaborator Contribution | Our collaborators have contributed their esxpertise in platelet biology, pulmonary inflammation and intravital microscopy. |
| Impact | Joint publication in Blood in 2015. Planned joint grant applications. Continued collaboration on related projects. |
| Start Year | 2012 |
| Description | Simon Pitchford and Clive Page (KCL, London) |
| Organisation | King's College London |
| Country | United Kingdom |
| Sector | Academic/University |
| PI Contribution | Role of platelets in leukocyte recruitment |
| Collaborator Contribution | joint experiments lab visits joint publications |
| Impact | joint papers in Blood and in Curr Opin In Hematol |
| Start Year | 2009 |
| Description | Till Bretschneider |
| Organisation | University of Warwick |
| Department | School of Life Sciences |
| Country | United Kingdom |
| Sector | Academic/University |
| PI Contribution | We collected imaging data describing neutrophil migration using a number of imaging modalities. |
| Collaborator Contribution | Data analysis |
| Impact | Publications and presentations at meetings |
| Start Year | 2010 |
| Description | Williams - PI3K |
| Organisation | Medical Research Council (MRC) |
| Department | MRC Laboratory of Molecular Biology (LMB) |
| Country | United Kingdom |
| Sector | Academic/University |
| PI Contribution | Intellectual input, reagents, personnel |
| Collaborator Contribution | Intellectual input, reagents, personnel |
| Impact | Angulo et al (2013) Science 342(6160):866-71. doi: 10.1126/science.1243292 Zhang et al (2011) Mol Cell. 41(5):567-78. doi: 10.1016/j.molcel.2011.01.026 Houslay et al (2016) Sci Signal. 9(441):ra82. doi: 10.1126/scisignal.aae0453 Krugmann et al (2004) Curr Biol.14(15):1380-4 |
| Description | Yonathan Lissanu-Deribe and Lynda Chin (Anderson Cancer Center, Texas University, USA): |
| Organisation | University of Texas |
| Department | M. D. Anderson Cancer Center |
| Country | United States |
| Sector | Academic/University |
| PI Contribution | Role of P-Rex2 in melanoma growth and metastasis |
| Collaborator Contribution | joint experiments |
| Impact | none yet |
| Start Year | 2012 |
| Description | Career Advice Lecture |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | Yes |
| Geographic Reach | Local |
| Primary Audience | Public/other audiences |
| Results and Impact | Lecture to Babraham students and Post Docs on career in academia Improved communication between post docs, more senior researchers and the Institute's executive committee. |
| Year(s) Of Engagement Activity | 2012,2014,2016 |
| Description | Career Advice Lecture |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | Yes |
| Geographic Reach | Local |
| Primary Audience | Public/other audiences |
| Results and Impact | lecture to students and postdocs on a career in bioscience Answered many questions from the audience who never had the chance to experience work in a Research Institute as opposed to University. |
| Year(s) Of Engagement Activity | 2009,2012,2014 |
| Description | Editorial board member Cell Rep |
| Form Of Engagement Activity | A magazine, newsletter or online publication |
| Part Of Official Scheme? | No |
| Geographic Reach | International |
| Primary Audience | Professional Practitioners |
| Results and Impact | Editorial board member of Cell Reports |
| Year(s) Of Engagement Activity | 2012,2013,2014,2015,2016,2017,2018,2019 |
| Description | European Phagocyte Workshop - Member of steering committee |
| Form Of Engagement Activity | A formal working group, expert panel or dialogue |
| Part Of Official Scheme? | No |
| Geographic Reach | International |
| Primary Audience | Professional Practitioners |
| Results and Impact | Steering committee of Annual European Phagocyte Workshop |
| Year(s) Of Engagement Activity | 2011,2012,2013,2014,2015,2016,2017,2018,2019 |
| Description | Interview with PBS USA |
| Form Of Engagement Activity | A press release, press conference or response to a media enquiry/interview |
| Part Of Official Scheme? | No |
| Geographic Reach | International |
| Primary Audience | Media (as a channel to the public) |
| Results and Impact | Interviewed about effect of Brexit upon UK science and innovation |
| Year(s) Of Engagement Activity | 2017 |
| Description | Invited lecturer to international meetings (average 2-3 per year) |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | International |
| Primary Audience | Professional Practitioners |
| Results and Impact | promoted discussions, collaborations scientific collaborations, joint grants and publications |
| Year(s) Of Engagement Activity | Pre-2006,2006,2007,2008,2009,2010,2011,2012,2013,2014,2015,2016,2017,2018,2019 |
| Description | Organisor/chair of international conference |
| Form Of Engagement Activity | Participation in an activity, workshop or similar |
| Part Of Official Scheme? | No |
| Geographic Reach | International |
| Primary Audience | Professional Practitioners |
| Results and Impact | FASEB meeting Small GTPAses FL June 2016, organizer and chair |
| Year(s) Of Engagement Activity | 2015 |
| Description | Phagocytes.Net |
| Form Of Engagement Activity | A magazine, newsletter or online publication |
| Part Of Official Scheme? | Yes |
| Geographic Reach | International |
| Primary Audience | Public/other audiences |
| Results and Impact | I am Editor of this website for fellow phagocyte researchers, postdocs, students and the general public that provides information on phagocyte-related events, research groups, and jobs. www.phagcytes.net Better communication among research groups in the phagocyte field. Markedly increased numbers of participants at the annual meetings of the European Phagocyte Workshop. |
| Year(s) Of Engagement Activity | 2006,2008,2009,2010,2011,2012,2013,2014,2015,2016,2017,2018 |
| URL | http://www.phagcytes.net |
| Description | Public feedback exercise |
| Form Of Engagement Activity | A formal working group, expert panel or dialogue |
| Part Of Official Scheme? | No |
| Geographic Reach | Regional |
| Primary Audience | Professional Practitioners |
| Results and Impact | The Signalling Department, in parallel with other BI departments, presented our research to a professionally selected sample of the UK public with the interntion of gaining public feedback on our institute and research. |
| Year(s) Of Engagement Activity | 2015 |
| Description | School visits in Cambridgeshire |
| Form Of Engagement Activity | A magazine, newsletter or online publication |
| Part Of Official Scheme? | Yes |
| Geographic Reach | Regional |
| Primary Audience | Schools |
| Results and Impact | talks at primary schools Raised image of "the scientist" among scholl chidren in the area over the past years (questionnaires). |
| Year(s) Of Engagement Activity | 2006,2008,2010,2011,2012,2013,2014,2015 |
| Description | Schools Days at Babraham |
| Form Of Engagement Activity | Participation in an open day or visit at my research institution |
| Part Of Official Scheme? | Yes |
| Geographic Reach | Local |
| Primary Audience | Schools |
| Results and Impact | We host 10 GCSE or A level students for one day every year to conduct experiments. Rasied image of "science" as a profession and of "scientists" among young people in the area. |
| Year(s) Of Engagement Activity | 2006,2007,2008,2009,2010,2011,2012,2013,2014,2015,2016,2017,2018,2019 |
| Description | Work experience for year 12 school students |
| Form Of Engagement Activity | Participation in an activity, workshop or similar |
| Part Of Official Scheme? | No |
| Geographic Reach | Local |
| Primary Audience | Schools |
| Results and Impact | Work shadowing for a yr 12 school student to understand biomedical research Student applied and got an offer on a biomedical degree course |
| Year(s) Of Engagement Activity | 2014 |
| Description | speaker/session chair at international conference |
| Form Of Engagement Activity | Participation in an activity, workshop or similar |
| Part Of Official Scheme? | No |
| Geographic Reach | International |
| Primary Audience | Professional Practitioners |
| Results and Impact | invited speaker and/or session chair at international conferences |
| Year(s) Of Engagement Activity | 2016,2017,2018 |