Signalling and biological roles of the class II and III PI 3-kinase enzymes

Lead Research Organisation: University College London
Department Name: Oncology

Abstract

This proposal seeks to investigate the roles and mechanism of action of a group of molecules, known as phosphoinositide 3-kinases (PI3Ks). When cells receive signals from the outside world, PI3Ks generate signals inside cells to make the cells respond. This output can be very diverse, and includes cell proliferation, survival and resistance against stress, but also the production of hormones, cell migration or even cell death. The PI3K pathway has been implicated in diseases such as cancer, inflammation, auto-immunity and diabetes. This pathway is therefore considered as an interesting new target for drug development. There are three subgroups of PI3K and thus far, scientists (including ourselves) have mainly focused on the so-called class I subset of PI3Ks. Drugs against the class I PI3Ks are currently being tested in clinical trials in human cancer. At the moment, very little is known about the class II and III PI3Ks, despite many years of research by others. We have in the past been very successful in finding out what the class I PI3Ks are doing and how they can be targeted in disease. We have now embarked on a scientific research programme to find the biological roles of the class II and III PI3K, and to identify how they transduce signals inside cells. We will do this by studying mice in which the class II or III PI3Ks have been inactivated, in order to uncover what these PI3Ks do in the living organism, and how they work. We have made some interesting initial observations, and now propose to progress to a full characterisation of these mice and the mechanism of action of these poorly-investigated members of the PI3K family. This is a fundamental science project that will enhance our knowledge about basic biological phenomena. In the past, the biology of PI3Ks has impacted on science far beyond our own field, mainly because these enzymes control fundamentals of biology. This proposal also has the potential to benefit industry, as it might identify PI3Ks as new targets to develop medicines against. In the longer term, it is very likely that this research may lead to a better understanding of disease processes and to the development of new medicines. This proposal is part of an ongoing, multidisciplinary research project with collaborators at five UK Universities and CRUK, as well as international collaborators.

Technical Summary

Phosphoinositide 3-kinases (PI3Ks) are a conserved family of lipid kinases which generate lipid second messengers inside cells. The PI3K pathway has been implicated in diseases such as cancer, inflammation, auto-immunity and diabetes. This pathway is therefore considered as an interesting new area for drug development. Mammals have 8 isoforms of PI3K, subdivided into three classes. While the class I PI3Ks have been extensively studied (by ourselves included), the roles and mechanism of signalling of the class II and III PI3Ks remain largely unknown. Indeed, despite thousands of publications on PI3K in the public domain, the physiological roles of the majority of PI3K isoforms remains unknown. This research proposal aims to characterise the roles of the class II and III PI3Ks in normal physiology, their integration in signalling by extracellular stimuli and their downstream signalling and cell biology, at the organismal and cellular level. We will hereby exploit new, gene-targeted mice that we have generated (unpublished). A first round of characterisation of these mice has yielded interesting biological observations that we now seek to understand at the molecular level. This is a multi-disciplinary project that covers animal physiology, embryology, cell biology, signal transduction and proteomic approaches, at the cellular and whole organism level.

Planned Impact

This proposal seeks to investigate the roles of a group of lipid kinases, PI3Ks, which we believe play important roles in cell signalling. We have created a panel of mice in which the genes for these kinases have been inactivated, and we wish to investigate these to delineate the roles of these PI3Ks in cell biology and signalling. In particular, we aim to decipher the molecular mechanism why some mice respond better to metabolic stimuli. This could elucidate the mechanisms underlying metabolic diseases and ultimately lead to the use of inhibitors of class II/III PI3Ks as insulin-sensitizers for the treatment of these diseases. Beneficiaries of the proposed research include the academic research community, the commercial private sector and the wider public, who will benefit as follows: - this is a cross-UK University collaboration which is expected to strengthen interactions within the sector. For the academic researchers, we expect that our research will open new interdisciplinary avenues for exploration, shedding light on a broad range of key cell biological phenomena. In the past, the biology of PI3Ks has impacted on science far beyond our own field, mainly because they control fundamentals of biology. Immediate academic beneficiaries will be the named senior RAs who are likely to exploit this to set up their own independent research teams, or become involved in drug development, through our extensive interactions with industry. - this proposal has the potential to benefit the commercial sector by identifying kinases as new targets for therapeutic intervention. The enzymes under study are potential targets for small molecule inhibitors, although the disease indication of such inhibitors is preliminary. - in the longer term, the wider public might also benefit from our research, in that it leads to a better understanding of disease processes, especially in the context of diabetes and obesity. More than 2.5 million people in the UK have been diagnosed with diabetes. Thus, the proposed project may lead in the longer term to the development of new medicines to cure these diseases, enhancing quality of life in the UK and worldwide. We believe that the timescales for the benefits to be realised are not too distant. In the academic context, impact will be within the time frame of the grant proposal. Indeed, we have already carried out substantial preliminary work in this area, and all research staff are in place, allowing us to progress at a fast pace. The research and professional skills that the staff on this project will develop are very broad. Other than being involved in high profile and cross-discipline research in multiple UK Universities, staff will also be trained in project management and communicating their science in the broadest sense, providing them with transferable skills beyond pure academic research. Plans to ensure that the beneficiaries will benefit are in place and can be summarised as follows: - the research outputs will be communicated to the academic community through traditional scientific communication routes including high-profile peer-reviewed publications and international scientific conferences. All applicants will also actively engage in efforts to promote the public understanding of the science underlying the proposed project. - the proposed partnerships are a formal extension of successful ongoing collaborations with track records in generating impact and increasing UK competitiveness. The majority of the work will be managed from the PI's laboratory, with clearly identified roles and responsibilities of all partners. - BV and co-applicants have ample experience in commercialising the output of their research. BV and PC hold several patents and have recently co-founded a new spin out company (Activiomics; Feb 2010). Through their broad network of contacts and interactions with industry, they are aiming to protect and exploit the IP generated from the proposed research.

Related Projects

Project Reference Relationship Related To Start End Award Value
BB/I007806/1 03/04/2011 05/01/2014 £1,161,839
BB/I007806/2 Transfer BB/I007806/1 06/01/2014 28/02/2015 £208,921
 
Description see BB/I007806/1 where I have added all this information - unfortunately, this grant has been split over two academic institutions due to our lab move during the time of the grant. This means that I would have to duplicate every entry. My apologies for not having done this.
Research is ongoing into the potential of PI 3K modulators for treating ischemia reperfusion injury (at present, no clinical treatments exist for this form of injury).
Exploitation Route see BB/I007806/1 where I have added all this information - unfortunately, this grant has been split over two academic institutions due to our lab move during the time of the grant. This means that I would have to duplicate every entry. My apologies for not having done this.
Sectors Healthcare

 
Description Using genetic approaches, we have discovered that inactivation of one of the kinases investigated in this grant application results in a remarkably improved glucose tolerance and insulin sensitivity in mice, with protection from high fat diet-induced hepatic steatosis. We have successfully applied for a BBSRC Follow-on Fund to pharmacologically validate these data.
First Year Of Impact 2013
Sector Healthcare,Pharmaceuticals and Medical Biotechnology
 
Description BBSRC Follow-on Fund
Amount £170,000 (GBP)
Organisation Biotechnology and Biological Sciences Research Council (BBSRC) 
Sector Public
Country United Kingdom
Start 01/2014 
 
Description Global Challenges Research Fund - Impact Acceleration Account
Amount £20,000 (GBP)
Organisation Biotechnology and Biological Sciences Research Council (BBSRC) 
Sector Public
Country United Kingdom
Start 09/2016 
End 03/2017
 
Title vps34 mutant mice 
Description Mice in which the vps34 PI 3-kinase is inactivated in a constitutive or conditional manner. This mouse line can be used in pre-clinical models of disease, such as cancer and diabetes. We may ultimately licence these mice to pharma, as we have done for our other PI3K mutant mice. 
Type Of Material Model of mechanisms or symptoms - mammalian in vivo 
Year Produced 2011 
Provided To Others? Yes  
Impact - Extensive collaborations in UK, US and France. - Allowed other people to apply for grants using these materials - successful bid to BBSRC for programme grant follow-up funding 
 
Description Collaboration with Novartis Research Institution - Boston USA 
Organisation Novartis Institutes for BioMedical Research (NIBR)
Country United States 
Sector Private 
PI Contribution We have received a small molecule inhibitor from Novartis under MTA, and tested this compound to validate our genetic data
Collaborator Contribution Novartis provided a small molecule inhibitor against the vps34 Pi 3-kinase under MTA,
Impact A manuscript describing the results of these studies has now been published (Bilanges et al - Nature Communications 2017)
Start Year 2016
 
Description Collaboration with Prof Volker Haucke Berlin - Leibniz-Forschungsinstitut für Molekulare Pharmakologie (FMP) 
Organisation Leibniz Association
Department Leibniz-Institute for Molecular Pharmacology
Country Germany 
Sector Academic/University 
PI Contribution Exchange of reagents, information and personnel. The Named Postdoctoral Fellow on this grant has secured a research position at the FMP in Berlin. This Lab continues research in this area of research, and will use the data obtained for further studies and to complete a publication.
Collaborator Contribution Intellectual contributions - we have a joint paper under review in Developmental Cell (to be resubmitted post revision in April 2022)
Impact No impact yet
Start Year 2020
 
Description Mass Spectrometry Facility at Barts Cancer Institute - Queen Mary University of London 
Organisation Queen Mary University of London
Country United Kingdom 
Sector Academic/University 
PI Contribution The Centre for Cell Signalling has set up a proteomics facility that has provided a service to Barts Cancer Institute.
Collaborator Contribution CRUK has contributed to the salary of a mass spectrometry technician, as part of the core support grant it provides to Barts Cancer Institute.
Impact Successful grant applications and collaborative papers
Start Year 2012
 
Description Measurement of phosphoinositides levels in cells and mice - Bernard Payrastre - France 
Organisation National Institute of Health and Medical Research (INSERM)
Department Institute of Metabolic and Cardiovascular Disease (I2MC)
Country France 
Sector Public 
PI Contribution Developing new genetic models with inactive PI 3-kinase isoforms
Collaborator Contribution Advanced lipid analysis techniques
Impact Joint publications are being prepared
Start Year 2012
 
Description University of Toulouse, France 
Organisation Paul Sabatier University (University of Toulouse III)
Country France 
Sector Academic/University 
PI Contribution The key contribution to this discovery for a role of PI3K-C2beta in stroke derives from the use of mice in which this kinase has been inactivated. Our investigations further identify a functional explanation for this finding, in that this kinase is imporant for how cells stick to their outside substrate or their neighbouring cells.
Collaborator Contribution Our Collaborators made the primary discovery by subjecting our mutant mice to experimental models of stroke. They also performed the lion share of the experimental work.
Impact 1. joint publication 2. IP filed
Start Year 2018
 
Title SCREENING METHOD 
Description The present invention provides a method for identifying agents useful in the treatment and/or prevention of a disease associated with insulin resistance and/or glucose intolerance which comprises the step of investigating the capacity of a test agent to inhibit the Vps34 signalling pathway and/or the RhoIota3Kappa-02beta signalling pathway. The present invention also provides a transgenic non-human animal which comprises a mutation in the gene encoding Vps34 or RhoIota3Kappa-C2beta such that the active site is inactivated. 
IP Reference WO2013076501 
Protection Patent application published
Year Protection Granted 2013
Licensed No
Impact Potential impact on insulin-resistant related disease
 
Title USE OF PI3KC2B INHIBITORS FOR THE PRESERVATION OF VASCULAR ENDOTHELIAL CELL BARRIER INTEGRITY 
Description Ischemic conditions are a leading cause of death for both men and women. Ischemia, a condition characterized by reduced blood flow and oxygen to an organ. Re-establishment of blood flow, or reperfusion, and re-oxygenation of the affected area following an pot ischemic episode is critical to limit irreversible damage. However, reperfusion also associates potentially damaging consequences. For NI instance, increased vascular permeability is an important contributor to edema and tissue damage following ischemic events. Here the inventors shows that genetic inhibition of PI3K-C2ß reduces cerebral infarction in two ischemia/reperfusion (UR) models and improves neurological outcome. The genetic inhibition stabilizes the blood-brain barrier (BBB) after ischemic stroke and reduces inflammation. Accordingly, the present invention relates to a method for the preservation of vascular endothelial cell barrier integrity in a patient in need thereof comprising administering to the subject a therapeutically effective amount of a PI3KC2ß inhibitor. 
IP Reference US2021238605 
Protection Patent application published
Year Protection Granted 2021
Licensed Commercial In Confidence
Impact Identifies a new drug target for the treatment and/or prevention of stroke
 
Description Lab Tours and School Visits 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? Yes
Geographic Reach Local
Primary Audience Public/other audiences
Results and Impact - over 200 lay people have attended tours in our laboratory
- PI presented the work to over 500 schoolchildren over the period 2007-2012
- PI has hosted placements of pupils in the lab


- PI is asked for return visits every year + additional invitations to talk
- stimulates children to consider careers in science and research
Year(s) Of Engagement Activity 2007,2008,2009,2010,2011,2012
 
Description Science Week Events and Talks 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Schools
Results and Impact Discussion on working in Science - animals in research

various interactions with people interested in science
Year(s) Of Engagement Activity 2006,2007,2008,2009,2010,2011,2012,2013