I Optimization Of the Nanosensors For Pyruvate And Lactate.Ii Development Of Methods For the High-throughput Screening Of Energy Metabolism Modulation In Cultured Cells.Iii Generation Of a Transgenic Mouse Line For Detection Of Altered Energy Metabolism.We Have Developed Genetically-encoded Fluorescent Nanosensors And Associated Methods For the Monitoring Of Cell Metabolism Using Microscopy. The General Objective Of the Present Project is To Transform Our Portfolio Of Metabolic Tools Into a Platform Of High-throughput Prototypes Of Commercial Interest. This Objective is In Register With the Aim Of the Center For Scientific Studies (cecs) Which is the Transformation Of Basic Research Knowledge Into Applications Of Economic Impact.The Center For Scientific Studies (cecs) Has Recently Developed Fluorescent Nanosensors For Metabolites And a Range Of Methods To Estimate Metabolism Using Microscopy. The General Objective Of This Project is To Integrate Our Portfolio Of Sensors And Associated Techniques Into a Platform Of Prototype Products And Services Of Commercial Interest. This General Objective is In Register With the Aim Of the Host Institution Cecs Which is the Transformation Of Basic Research Knowledge Into Applications Of Economic And Social Impact. This Project is Focused Towards the Pharmaceutical Industry Which Despite Its Continued Growth Over the Last Decades is Facing a Crisis. New Drugs Are Inordinately Expensive Because Of Low Industry Productivity the Average New Molecular Entity Requiring An Investment Of About Us$ 1 Billion Over a Period Of 10 - 15 Years. Discovering And Developing a Drug is a Hierarchical Process That Starts With Libraries Of Thousands Of Chemicals Which Are Screened For a Target Activity With An In Vitro Assay. Hits Are Promoted To Subsequent Stages the Testing Of Activity And Toxicity In Cells Tissues Animal Models And Then To Clinical Trials Where Most Candidates Fail As They Are Found To Be Ineffective In the Biological Context Or Show Untoward Side Effects. Attrition At Later Stages Of the Development Chain is the Most Important Determinant Of Overall R&d Efficiency. One Effective Manner To Increase the Number Of New Drugs And Reduce the Costly And Inefficient Use Of Animals is the Inception Of Intermediate Levels Of Screening Based On Cultured Cells Or Tissues Both To Confirm the Biological Activity Of a Compound Detected In Vitro And To Rule Out Cell-level Toxicity As Early As Possible. Cecs Has Recently Developed Fluorescent Nanosensors For the Intracellular Monitoring Of Metabolite Concentration And Flux With High Resolution Using Microscopy. Being Genetically Encoded These Nanosensors Are Amenable For High-throughput Implementation As Required For Efficient Drug Development But the Multiwell Plate Readers That Are Used For Drug Screening Are Less Sensitive Than High End Microscopes. Therefore To Be Commercially Attractive the Sensors Need To Be Optimized. We Propose Three Specific Objectives: Optimization Of the Nanosensors For Pyruvate And Lactate Whereby the Nanosensors For Pyruvate (pyronic Pct/ib2013/056766) And Lactate (laconic Pct/us2012/033639) Will Be Enhanced And Diversified In Terms Of Spectrum Dynamic Range And Affinity. Ii Development Of Methods For High-throughput Screening Of Energy Metabolism Modulation In Cultured Cells Which Implies the Translation Of Microscope-based Techniques For the Monitoring Of Metabolite Concentrations And Fluxes (pct/us2010/40643) Into Prototypes For High-throughput Assays Using Multiwell Plate Readers And Iii Generation Of a Transgenic Mouse Line For Detection Of Altered Energy Metabolism For Preclinical Studies Which Will Take Advantage Of Our In-house Experience On Genetic Modification. The Project is Planned Over 36 Months And Runs Along Three Interconnected Lines Of Work. The Backbone is the Continuous Optimization Of the Nanosensors. As Improved Variants Are Produced They Will Be Tested In the Parallel Lines Conducive To Prototypes. Altogether the Sensors And Methods Developed And Optimized During the Project Will Constitute a Platform Of Tools To Be Offered To Pharmaceutical And Biotech Companies In the Way Of Patent Licenses Products And Services. We Envisage That the Utilization Of High-throughput Methods For the Screening Of Metabolism Will Help Discovering New Drugs And Reduce Their Cost Impacting Favorably On Healthcare.Corporación de Fomento de la Producción