ENIGMAS · Explicitly Normalized ImaGing MAss Spectrometry
7РП — „Хора“ (Действия „Мария Кюри“)
- Период
- 2012-10-01 → 2014-09-30
- Финансиране от ЕС
- 183 806 €
- Участници
- 1
- Схема
- MC-IEF
Линиите свързват координатора с партньорите.
Накратко на български
Методите за мас-спектрометрично изобразяване се разработват за анализ на протеини и метаболити в мозъка, например при проучване на мигрената. Тези инструменти помагат за по-доброто разбиране на неврологичните заболявания чрез използването на животински модели.
Кратко обяснение, генерирано от езиков модел по текста на CORDIS. Оригиналът е по-долу.
Резултати накратко
Explicitly Normalized ImaGing MAss Spectrometry
The Explicitly Normalized ImaGing MAss Spectrometry (ENIGMAS) project aimed at developing methodologies for imaging mass spectrometry (MS) to enable its application to the study of pre-clinical animal models of neurological disease. Main developments/achievements include: - Reproducible on-tissue digestion protocols for protein analysis in imaging MS. - Reproducible sample preparation protocols for metabolite, neuropeptide and intact protein analysis by imaging MS. - Comprehensive database of peptides and proteins identified by imaging MS in mouse brain tissue sections. - Bio informatics pipelines for automated and anatomical driven analysis of imaging MS datasets from large animal cohorts. - Application of imaging MS to pre-clinical models of neurological disease. - Novel biomolecular insights regarding the changes produced in mouse brain by cortical spreading depression, the electrophysiological equivalent of migraine aura in animals. The work developed during the grant period was enthusiastically disseminated across the imaging MS and neuroscience communities through the publication of peer reviewed manuscripts (2 published manuscripts, 1 submitted manuscript, 1 manuscript in preparation), participation in scientific conferences (oral and poster communications), participation in workshops and advanced training across Europe through the COST programme BM1104 – Imaging Mass Spectrometry: New Tools For Healthcare Research. The technological improvements will be made publically available through the imaging MS website (maldi-msi.org) for widespread application across different imaging MS laboratories. The innovative technologies/methodologies developed will have a profound impact on the imaging MS and on the neurosciences field. Most important, they will allow: - Facile protein identification through the developed imaging MS methods and protein databases; - Automatic annotation of imaging MS datasets with the tissue histology/anatomy; - Facile comparison of animal cohorts in pre-clinical models of neurological disease, whether by extracting data from specific regions in the mouse brain or comparing distributions in different animals; - Alignment of imaging MS datasets to the Allen Brain Atlas, a genome-wide repository of data from the mouse brain, allows the comparison of imaging MS data with genomic information, thereby increasing the impact of imaging MS data and biological information retrieved; - Widespread application of imaging MS to other neurological diseases of present day importance, such as stroke, seizure, Alzheimer’s, Parkinson’s.
Текст от CORDIS, на английски · Данни: CORDIS, © Европейски съюз
Цел на проекта
Imaging Mass Spectrometry is a rapidly developing technique that uses spatially resolved proteomic techniques to investigate a tissue’s biomolecular content. Imaging MS of pharmaceuticals has demonstrated that relative quantitation can be obtained if the MS response factors are known. However MS response factors for peptides and proteins are highly dependent on their exact nature. Despite it being acknowledged as the single biggest factor limiting the clinical application of the technique, to date it has remained an enigma how imaging MS analyses may be explicitly normalized to enable relative quantitation because it would require the relative response factors to be determined for every peptide/protein.Here I will develop a methodology for the explicit normalization of every protein detected during imaging MS experiments. This will utilize the recently released SILAC mouse to create a reference standard in which all lysine residues are replaced with 13C6-lysine. Tryptic digestion of SILAC protein extract creates tryptic peptides containing a single 13C6-lysine, which can be used as a reference standard for the 12C6-lysine containing peptides created by on-tissue digestion of normal mouse tissue.Ultrahigh mass resolution imaging MS will be used to ensure all tryptic peptides, and their 13C6-lysine reference standards, are resolved enabling the images of all tryptic peptides to be normalized using their own isotopically labelled analogues. The quantitative capabilities will be determined for every tryptic peptide ion detected from mouse brain tissue.The capabilities of this ENIGMAS strategy (Explicitly Normalized ImaGing Mass Spectrometry) will be used to investigate the spatial and temporal evolution of the proteome perturbations that follow migraine. Multiple measurement repeats and multiple animals in each group will provide the first measure of statistical significance for imaging MS based molecular histology, a crucial step toward its clinical application.""
Оригинален текст от CORDIS (на английски).
Участници
- ACADEMISCH ZIEKENHUIS LEIDEN · LeidenКоординаторНидерландия
Връзки
Данни: CORDIS, © Европейски съюз