CHEMOFLUXOME · Development of metabolomics and fluxomics methods for metabolic drug target and toxicity elucidation using yeast
6РП — Действия „Мария Кюри“
- Период
- 2007-01-01 → 2008-12-31
- Финансиране от ЕС
- 173 831 €
- Участници
- 1
- Схема
- EIF
Линиите свързват координатора с партньорите. За проекти отпреди 2014 г. CORDIS не винаги дава точни координати. Тези точки са на ниво град или държава.
Накратко на български
Метаболитните процеси в дрожди се анализират чрез измерване на концентрацията на вещества и скоростта на химичните реакции при воздействие с лекарства. Новият бърз метод помага за по-точното определяне на токсичността на лекарствата и начина, по който те влияят на организма.
Кратко обяснение, генерирано от езиков модел по текста на CORDIS. Оригиналът е по-долу.
Резултати накратко
Final Activity Report Summary - CHEMOFLUXOME (Development of metabolomics and fluxomics methods for metabolic drug target and toxicity elucidation using yeast .)
The ability to assess global gene expression of drug-treated yeast cells has greatly contributed to uncover the mode of action of bioactive compounds. Altered levels of gene expression however, do not monitor the actual biological function. For a more direct functional readout of the large set of metabolic functions, we determine here intracellular fluxes (i.e. fluxome) and metabolite concentrations (i.e. metabolome) to screen drug interactions with the complex metabolic network of the model eukaryote Saccharomyces cerevisiae. As a proof of concept, we profiled dose dependent effect of 41 drugs with known specific targets in metabolism that are highly relevant for metabolic disease and toxicity. To achieve high-throughput metabolome analysis, we developed a rapid quenching and extraction procedure for microtiter plate-grown cells. Using direct sample injection, we then (semi) quantified 55 intracellular metabolites within central metabolism by ESI-MS/MS analysis. Additionally, metabolic fluxes were calculated from GC-MS-detected 13C-pattern in protein-bound amino acids. The results demonstrate that the metabolic function - monitored as fluxes - remains relatively robust to the drug perturbations, but that this robustness is either achieved by or is not yet affected by significantly altered pattern of intracellular metabolite concentrations. As the key result, we develop here a new high-throughput method for sensitive metabolomics. To the best of our knowledge, this is the first HT method for INTRACELLULAR metabolomics and it is at least 100-fold faster than any other global omics method that we are aware of. Our biological results demonstrate that this method has great potential in functional drug toxicity testing because we can effectively differentiate drugs with primarily local, specific metabolic responses and those with global, mostly off-target responses.
Текст от CORDIS, на английски · Данни: CORDIS, © Европейски съюз
Цел на проекта
This project addresses a key issue in pharmaceutical research from a fundamental technological perspective: functional drug target identification and early detection of toxic side effects. When assessing metabolic functions, current methods such as proteomics or transcriptomics have inherently indirect readouts and are often not representative of the in vivo metabolic state. The aim of this multidisciplinary project is to monitored functional drug interactions with components in complex metabolic networks b y applying and developing metabolomics and fluxomics methods in combination with multivariate statistics and machine-learning. Thereby, we will conceptually extend the capability to observe metabolic network operation well beyond the limits of current technologies.As a proof-of-concept, we focus on small molecule drugs that interfere with metabolism or its overlaying regulatory control network in the model eukaryote Saccharomyces cerevisiae. The hypothesis to be tested is whether multivariate statistical discrimination of mass spectrometry-detected nutritionally, pharmaceutically, or toxicologically relevant metabolic states have potential to conceptually improve early drug development. If successful, the novel approach is relevant for early in vivo characterization of drug target physiology and toxicology. While unicellular yeast is a pilot case, the approach can principally be extended to multi-cellular organisms and organs. Thus, this work has potential to improve the overall efficiency of European pharmaceutical research to treat human disease.The project will be carried out at the ETH Institute of Molecular Systems Biology and the Centre for Systems Physiology and Metabolic Diseases under the supervision of Prof. U Sauer. The Centre offers a multidisciplinary research environment with world-class expertise in metabolomics, computer science, cell biology and pathology.
Оригинален текст от CORDIS (на английски).
Участници
- EIDGENOSSISCHE TECHNISCHE HOCHSCHULE ZURICH · ZURICHКоординаторНиво градШвейцария
Връзки
Данни: CORDIS, © Европейски съюз