Babraham Institute develops new way of seeing signalling molecules at work
Collaboration between academic and commercial groups at the Babraham Institute – catalysed by facilities funded by the Biotechnology and Biological Sciences Research Council (BBSRC) – has developed a technique that will radically improve the ability to measure important cellular metabolite in cells and tissues, which could have applications in the development of new drugs.
The chemical in question, phosphatidylinositol 3,4,5-trisphosphate - better known as PIP3 or PtdIns(3,4,5)P3 - is a phospholipid in cell membranes that has an important role as a ‘messenger’, conveying signals between hormones outside the cell into critical intracellular functions such as cell growth, metabolism, survival and movement.
PIP3 is made in cells by a family of enzymes called phosphoinositide 3-kinases, commonly known as PI3Ks, of which there are different isoforms; these are activated by many groups of cell surface receptors to accelerate the production of PIP3. It is thought that identifying the appearance of PIP3 under the influence of different PI3K isoforms may bring new understanding of the class I PI3K signalling pathway and potential therapeutic interventions.
Professor Michael Wakelam, Director of the Babraham Institute and one of the paper’s authors explained, “The different isoforms are known to have distinct roles in maintaining health and pathology. Some play a critical role in the inflammatory response, some in how the body responds to insulin and others in oncology for example, so PI3Ks are currently among the most hotly pursued drug targets in the pharmaceutical industry.” This landmark methodology, published in Nature Methods, provides a means of monitoring a much needed biomarker for small molecule inhibitors of PI3Ks.
“Such inhibitors are currently being developed by several companies in the UK and abroad as novel therapeutics in the broad areas of oncology and inflammation,” said Dr Phill Hawkins, joint senior author of the paper. “This new technique may therefore offer powerful insights to the drug discovery process.”
Until recently methods for measuring PIP3 have been indirect, insensitive or very laborious and generally unable to discriminate between different fatty-acyl species. This new technique, however, heralds a significant advance in methodology for determining changes in this signalling molecule in cells. It is based on a chemical derivatisation of PIP3 that allows it to be very sensitively and accurately quantified by mass spectrometry. This allows PIP3 levels to be measured in extracts prepared from small samples of cells and tissues with relative ease. Furthermore, because it is the intact molecule that is measured, this method for the first time yields the variation in fatty acid composition of PIP3 in cells and hence paves the way to discovering how this might be important for PIP3 function.
A recent investment of £1.5M by BBSRC has enabled a significant enhancement of the Institute’s Mass Spectrometry Facility with the purchase of four new mass spectrometers, which are used to analyse almost any type of biomolecule including proteins (‘proteomics’) and lipids (‘lipidomics’). Babraham researchers in collaboration with other BBSRC-supported Institutes will use the Facility to analyse the importance of lipid signalling molecules in the immune system, exploring the basis of diet-related ill health, as well as in areas of crop research and food security.
Professor Maurice Moloney, Director of Rothamsted Research said, “One of the problems is that plant scientists and animal cell scientists don’t talk enough, but when we do, we find out all kinds of ‘unity of biology’ ideas. Babraham is way ahead in things like signalling with sphingolipids and Rothamsted Scientists are very interested from the plant side. So we are looking at how we can join forces in these matters.”
Dr Simon Carding of the Institute of Food Research (IFR), an Institute of BBSRC based in Norwich, will be using the facility to investigate the interplay between the intestinal immune system and metabolism. This relates to the ability of specific immune cells, called dendritic cells, to influence fat uptake and metabolism – key drivers in the pathophysiology of metabolic syndrome and obesity. “The current buzzword for this work is immunometabolism,” he said. “Our research, in collaboration with Professor Stella Knight of Imperial College London, will investigate how uptake of fat by dendritic cells affects their ability to initiate and regulate immune responses. These projects will rely on the lipidomic capability of the Babraham Institute.”
The Babraham Institute, an institute which receives strategic funding from the BBSRC, also plans to make this method and ‘enabling technology’ available for outside academic and commercial collaboration on a fee-for-service basis, resonating with BBSRC’s commitment to develop strategic partnerships and make available facilities to HEIs and the life science industry.
This work was supported by the Biotechnology and Biological Sciences Research Council (BBSRC) and the European Union. The research is also a collaboration between three Babraham Institute research groups (Hawkins, Stephens and Wakelam) and chemists in the Technology Development Laboratory (TDL), which was established in 2008 by the Institute’s commercial arm - Babraham Bioscience Technologies Ltd – to support innovation in bioscience fields.
Clark J, Anderson KE, Juvin V, Smith TS, Karpe F, Wakelam MJO, Stephens LR, Hawkins PT (2011) Quantification of PtdInsP3 molecular species in cells and tissues by mass spectrometry. Nature Methods 8 267-272 http://dx.doi.org/10.1038/nmeth.1564
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About the Babraham Institute:
The Babraham Institute undertakes world-class life sciences research to generate new knowledge of biological mechanisms underpinning ageing, development and the maintenance of health. Our research focuses on cellular signalling, gene regulation and the impact of epigenetic regulation at different stages of life. By determining how the body reacts to dietary and environmental stimuli and manages microbial and viral interactions, we aim to improve wellbeing and support healthier ageing. The Institute is strategically funded by the Biotechnology and Biological Sciences Research Council (BBSRC), part of UK Research and Innovation, through an Institute Core Capability Grant and also receives funding from other UK research councils, charitable foundations, the EU and medical charities.
The Biotechnology and Biological Sciences Research Council (BBSRC) is the UK funding agency for research in the life sciences. Sponsored by Government, BBSRC annually invests around £450 million in a wide range of research that makes a significant contribution to the quality of life for UK citizens and supports a number of important industrial stakeholders including the agriculture, food, chemical, health and well-being and pharmaceutical sectors. BBSRC carries out its mission by funding internationally competitive research, providing training in the biosciences, fostering opportunities for knowledge transfer and innovation and promoting interaction with the public and other stakeholders on issues of scientific interest in universities, centres and institutes.