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Sharing the fat of the land
Cross-institute collaborations to make most of new lipidomics facilities
21 March 2011
Working together is often better than working alone, and collaborations between BBSRC-funded institutes are set to prove that the whole is greater than the sum of the parts – especially when it comes to utilising high-tech equipment to solve problems that require ultra sensitive measurements of cellular metabolites in both animals and plants.
A recent £1.5M investment by BBSRC to purchase of four advanced mass spectrometers for the Babraham Institute's (BI) Mass Spectrometry Facility has already led to the publication of a new methodology (ref 1) in Nature Methods, which follows other papers in lipid analysis.
In addition to Babraham's existing collaborative lipidomics research with UK universities and industry, Babraham researchers will now be collaborating with other BBSRC-supported institutes and use the facility to analyse lipid signalling molecules in the immune system, to explore the basis of healthy diets, and to investigate other areas in crop research and food security.
One of four advanced ABI QTRAP4000 mass spectrometers (beige, right) with an attached Waters Ultrapure Liquid Chromatography system (blue, left) at the Babraham Institute.
Image: Babraham Institute
It pays to talk
Professor Maurice Moloney, Director of Rothamsted Research (RRes), an Institute of BBSRC, says that during talks with Babraham's Director, Michael Wakelam, he realised there was great scope for collaboration. "It came to light that not only do they do world-class work on phospho-inositide signalling, which is highly relevant for our work on crop plants, but that there is a state-of-the-art lipidomics group there too."
Moloney says that BI is way ahead in fields such as the measurement of certain fats called sphingolipids, and a group at Rothamsted are very interested in the workings of this molecule from the plant side. "So we are looking at how we can join forces in these matters," says Moloney. "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."
It's work that will be taken forward by Moloney's colleague Professor Jonathan Napier. He says the fact that they are both using the same platform instrument (an ABI QTRAP4000, pictured above) should help them share methods and standards. "Given that we have recently done some work on lipidomics in knockout mice (ref 2) I think there is potential for significant interactions," says Napier.
Taking the lead
Teamwork certainly lies ahead, but that hasn't stopped BI taking the lead with the new facilities and formulating an exciting new method to measure an important cellular metabolite in cells and tissues which could spur the development of new drugs.
Researchers at Rothamsted will share expertise and calibration data with collaborators at Babraham. Image: Rothamsted Research
The chemical in question, PIP3 (phosphatidylinositol 3,4,5-trisphosphate, or PtdIns(3,4,5)P3), is a phospholipid in cell membranes that conveys signals between hormones outside the cell into critical intracellular functions such as cell growth, survival and movement.
PIP3 is made in cells by a family of enzymes, PI3Ks (phosphoinositide 3-kinases), and BI Director Michael Wakelam says the different PI3K isoforms are known to have distinct roles in maintaining health and pathology. "Some play a critical role in the inflammation response, others in oncology. Hence the PI3Ks are currently among the most hotly pursued drug targets in the pharmaceutical industry."
This landmark methodology (ref 1), published in Nature Methods, provides a means of monitoring a biomarker for 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," says Dr Phill Hawkins, joint senior author of the paper and also based at Babraham. "Therefore this new technique may offer powerful insights to the drug discovery process."
Until recently, methods for measuring PIP3 have been indirect, insensitive or very laborious. The new technique heralds a significant advance in methodology for determining this signalling molecule in cells. It is based on a chemical derivation of PIP3 that allows it to be very sensitively and accurately quantified by mass spectrometry. This allows PIP3 levels to be measured in small samples of cells and tissues with relative ease and, for the first time, yields the variation of PIP3 in cells, paving the way to discovering how this might be important for PIP3 function.
PIP3 activates downstream signalling pathways required for cell growth and survival
BI plans to make the new method an enabling technology available for outside academic and commercial collaboration, fulfilling BBSRC's commitment to develop strategic partnerships with higher education institutions and the life science industry.
The research, supported by BBSRC and the European Union, was itself a collaboration between three BI research groups and chemists in the Technology Development Lab (TDL) which was established in 2008 by BI's commercial arm, Babraham Bioscience Technologies Ltd, to support innovation in bioscience fields.
Still in the animal world, another cross-institute use of Babraham's new facilities will investigate the interplay between the intestinal immune system and metabolism and 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," says Dr Simon Carding of the University of East Anglia and the Institute of Food Research (IFR), an Institute of BBSRC, based in Norwich, who will undertake the studies at IFR in parallel with collaborator Professor Stella Knight of Imperial College who is based at St. Mark's Hospital in Harrow.
Extending the collaborative aspect of Babraham's research and facilities further, a second aspect to Carding's work is being carried out by Knight's group which will investigate how uptake of fat by dendritic cells affects their ability to initiate and regulate immune responses. "Both of these projects will rely on the lipidomic capability of the Babraham Institute," says Carding.
A measure of the power of the facilities to measure fundamental principles common to all biochemical processes is that they can be used in the plant and animal kingdoms.
Plant scientists trying to enhance food security by improving crop plants have noted a signalling molecule of emerging significance, trehalose-6-phosphate (T6P), a sugar increasingly seen as important in determining agriculturally important traits.
Tinkering with T6P pathways in model plant Arabidopsis has revealed its importance.
T6P is a critical component of mechanisms that coordinate metabolism with growth and development – in Arabidopsis disruption of the first step of trehalose synthesis has lethal consequences and genetic modification of the trehalose pathway produces dramatic changes to the forms and shapes of plants – traits that could be used to tailor useful crop traits.
"It [T6P] is a real tough one to measure accurately," says Moloney. "But again, talking with Michael at Babraham it became clear that with all the cell-to cell communication work at BI, specifically their work on inositides, the methodologies sugar phosphates are well worked out and so we hope to find short-cuts to our technical problems that BI may already have in-hand through its previous work."
"My bottom line is that plant biologists don't get out enough. It only took 30 minutes of conversation with Michael to realise how many things of mutual interest are going on between our two institutes."
- Quantification of PtdInsP3 molecular species in cells and tissues by mass spectrometry (external link)
- ELOVL2 controls the level of n-6 28:5 and 30:5 fatty acids in testis, a prerequisite for male fertility and sperm maturation in mice (external link)
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