P13K signalling; the rules of engagement
Lead Research Organisation:
Babraham Institute
Department Name: UNLISTED
Abstract
Abstracts are not currently available in GtR for all funded research. This is normally because the abstract was not required at the time of proposal submission, but may be because it included sensitive information such as personal details.
Technical Summary
Nearly all of the cells of our bodies contain three related proteins called PI3K alpha, PI3K beta and PI3K delta. We now know these PI3Ks play a very important role in transmitting information from hormones outside of the cell to the inside of cells. This information is needed for the cell to co-ordinate various complex responses to the hormone, such as cell growth, survival or movement. The key aim of this project is to define the roles for PI3Ks in a type of white blood cell called a neutrophil.
Planned Impact
unavailable
People |
ORCID iD |
| Len Stephens (Principal Investigator) |
Publications
Gyori D
(2017)
Class (I) Phosphoinositide 3-Kinases in the Tumor Microenvironment.
in Cancers
Gyori D
(2018)
Compensation between CSF1R+ macrophages and Foxp3+ Treg cells drives resistance to tumor immunotherapy.
in JCI insight
Hawkins PT
(2016)
Emerging evidence of signalling roles for PI(3,4)P2 in Class I and II PI3K-regulated pathways.
in Biochemical Society transactions
Kulkarni S
(2011)
PI3Kß plays a critical role in neutrophil activation by immune complexes.
in Science signaling
Luff DH
(2021)
PI3Kd Forms Distinct Multiprotein Complexes at the TCR Signalosome in Naïve and Differentiated CD4+ T Cells.
in Frontiers in immunology
Malek M
(2017)
PTEN Regulates PI(3,4)P2 Signaling Downstream of Class I PI3K.
in Molecular cell
Malek M
(2017)
PTEN Regulates PI(3,4)P2 Signaling Downstream of Class I PI3K.
in Molecular cell
Stephens L
(2011)
Signalling via class IA PI3Ks.
in Advances in enzyme regulation
Tsolakos N
(2018)
Quantitation of class IA PI3Ks in mice reveals p110-free-p85s and isoform-selective subunit associations and recruitment to receptors.
in Proceedings of the National Academy of Sciences of the United States of America
Vanhaesebroeck B
(2012)
PI3K signalling: the path to discovery and understanding.
in Nature reviews. Molecular cell biology
| Description | We have established that it is possible to use a genetic approach in mice to isolate specifc pi3k complexes from mouse tissues. previously this had proved very difficult because of technical issues with the use of bacterial DNA in mammalian cells, that we have now solved. We have created a set of mice in which all class IA PI3K subunits that are widely expressed are endogenously avi-tagged. These have drive publications and on-going follow-on work in our lab funded by the MRC on prostate cancer and have been supplied to a number of other "international " labs to support their work. Eg Sabitini lab (Salk). |
| Exploitation Route | This approach allows use of avi-tagging technology in mice to isolate any protein that is placed in frame with an avi-tag (15aa). It uses transgenically expressed BirA with mammalised codon usage (ROSA locus) to achieve efficient biotinylation of the tag in all mouse tissues tested. It has already been exported to a number of ther labs and underpins a lot of further work at BI on other funding. The apporach is very powerful and we are discussing further application with companies. We have obtained follow-on funding from the MRC to develop promising results on prostate cancer progression. |
| Sectors | Pharmaceuticals and Medical Biotechnology |
| Description | They have been used by companies interested in selectively inhibiting class IA PI3K signalling pathways in cancer. We have also shown how class I PI3Ks can contribute to regulatory control of T cell attack on tumours and that this can combine with CSF1 signalling to support tumour progression . Dual inhibition of class IA PI3K and CSF1 signalling can lead to synergistic inhibition of tumour progression in some models. |
| First Year Of Impact | 2017 |
| Sector | Pharmaceuticals and Medical Biotechnology |
| Impact Types | Economic |
| Description | MRC MICA project grant |
| Amount | £950,000 (GBP) |
| Organisation | Medical Research Council (MRC) |
| Sector | Public |
| Country | United Kingdom |
| Start | 01/2018 |
| End | 01/2022 |
| Title | Creation of PI3K avi-tag mice |
| Description | We introduced a 15 amino-acid Avi-tag into the C-terminus of endogenous genes encoding regulatory (p85alpha or p85beta) or catalytic (p110alpha, p110beta or p110delta) subunits of the Class IA PI3K family in mice. We also engineered expression of an optimised BirA (prokaryotic biotin ligase) expression cassette into the ROSA locus in mice and interbred these with the above Avi-tag mice. These mouse strains allow isolation of biotinylated Class IA PI3K subunits from mouse tissues and cell lines derived from them. |
| Type Of Material | Model of mechanisms or symptoms - mammalian in vivo |
| Year Produced | 2018 |
| Provided To Others? | Yes |
| Impact | First publications: PMID: 30442661 |
| Title | Mass_spectrometry_analysis_of_phosphoinositides |
| Description | In collaboration with Jonathan Clark in the Biological Chemistry Facility at the Babraham Institute we have developed novel mass spectrometry approaches for measuring different molecular species of phosphoinositides in cells and tissues. This work has utilised chemical derivatisation coupled to LC-MS and involved the synthesis of several isotope-enriched internal standards. |
| Type Of Material | Technology assay or reagent |
| Year Produced | 2011 |
| Provided To Others? | Yes |
| Impact | The main impacts have been numerous publications (>30 as of 2021), collaborations and clinical trials of PI3K inhibitors. |
| URL | https://www.babraham.ac.uk/science-services/biological-chemistry/jonathan-clark |
| 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 | Klaus_Okkenhaug_Avi-tag_mice |
| Organisation | University of Cambridge |
| Department | Department of Pathology |
| Country | United Kingdom |
| Sector | Academic/University |
| PI Contribution | We supplied Klaus Okkenhaug's laboratory with mice in which the gene encoding PI3Kdelta carries a C-T Avi-tag and which also expressed a BirA transgene. We also provided practical advice on how to isolate biotinylated-PI3Kdelta complexes from mouse lymphocytes. |
| Collaborator Contribution | They lead the project, providing the intellectual framework and the majority of the data. |
| Impact | Publication: PI3Kd Forms Distinct Multiprotein Complexes at the TCR Signalosome in Naïve and Differentiated CD4+ T Cells. doi: 10.3389/fimmu.2021.631271 |
| Start Year | 2018 |
| Description | Steve Shuttleworth |
| Organisation | Karus Therapeutics |
| Country | United Kingdom |
| Sector | Private |
| PI Contribution | We have performed research with neutrophils and xenografts and PI3K inhibitors and genetically modified mice to study the role of PI3Ks in immune function in inflammation and tumours. |
| Collaborator Contribution | They have provided funding and inhibitors. |
| Impact | See other outputs, some of the data is provided in the from of reports to Karus, some is published. |
| Start Year | 2012 |
| 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 | 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 |