Growing evidence indicates that severe derangements in lipid fingerprints are characteristic of many brain diseases and nervous system disorders including Alzheimer?s disease (AD), traumatic brain injury (TBI), spinal cord injury (SCI) and schizophrenia. There are many questions, however, regarding the nature of these derangements including how they occur, and whether therapies might be developed to correct or exploit them. Lipidomics is an important and timely area of research because of its ability to comprehensively and quantitatively evaluate levels of lipids responsible for signaling such as phosphoinositol metabolites and arachidonic acid metabolites providing insights into biology not previously available. This information when integrated with other ?omics? data in advanced bioinformatics models provides a superb method for identifying key drivers in biology and disease mechanisms, as well as identifying new drug targets for pharmaceuticals development. Advances in electrospray ionization-mass spectrometry have facilitated the precise profiling of lipid species within tissues, including low abundance lipid classes that participate in cell signaling and membrane trafficking attributable to disease pathogenesis. We propose to perform complete analysis of lipid fingerprints using liquid chromatograph mass spectrometry by LC/MS in combination with HPLC analysis within JJP VAMC expertise for lipidomics and metabolomics measurements from lipid extracts of cultured cell lines, primary cultures or tissue samples. The instrument requested in this application is a Triple Quadrupole LC/MS System with Jet Stream Technology consisting of high speed pumps, multiple columns (up to 8) with thermostat control between 4-1100C, multi-sampler up to 600 bars for well-plates and individual sample containers suitable for larger high throughput and pilot studies, respectively. The high throughput automation capabilities will let us analyze samples in 96-well plates in less than ten minutes which provide capacity for high throughput analysis of large scale samples. The instrument is designed to be easily used by individuals who do not have specialized training in lipidomics and metabolomics. These instruments are manufactured by Agilent Technologies. The purchase price includes on-site installation and familiarization training and a one-year warranty including a one-year PC image recovery and software upgrade and phone assistance. Data analysis software is available from the manufacturer for licenses of two workstations. The instrument?s detection limits are lower than ever and for the first time achieve zeptomole level sensitivity at convenient flow rates. No equivalent instrument dedicated to lipidomics and metabolomics is available at JJPVA or our academic affiliate the Icahn School of Medicine at Mount Sinai. If funded, the instrument will be used in VA-funded projects in the fields of Alzheimer?s disease, traumatic brain injury, and spinal cord injury. The instrument would add the capacity for systematic study of lipid homeostasis and metabolism in cultured cells, mouse and postmortem human brain and spinal cord tissue samples, and would thereby substantially broaden and enhance the understanding of medical conditions that affect the health and quality of life of veterans impacted by these diseases and disabilities.
Growing evidence indicates that severe derangements in lipid fingerprints are characteristic of many brain diseases and nervous system disorders including Alzheimer?s disease, traumatic brain injury, spinal cord injury and schizophrenia. Lipidomics comprehensively and quantitatively evaluate levels of complete lipid profiles responsible for signaling and other physiological function. This information when integrated with other ?omics? data in advanced bioinformatics models provides a superb method for identifying key drivers of diseases and new drug targets. If funded, this application would make available to JJPVA investigators a state of the art Triple Quadrupole LC/MS Detection System by which one can quickly and precisely determine changes in lipidomic fingerprints of living cells in the setting of neurodegenerative processes, injury or trauma. Such information will advance the understanding and treatment of health problems affecting veterans including dementia, traumatic brain injury, and spinal cord injury and other disabilities.
