Fig 11. The metabolic study platform.

Rats received MCT at day 0 and developed severe PH at day 28. However, lungs were collected for metabolomics at day 14 to monitor metabolic changes that precede pathological changes in the lung and heart. Lungs were perfused, collected and shipped to Metabolon inc for metabolic profiling. Sample preparation was done at Metabolon as described in the methods. Data from LC/GC MS were obtained and analyzed using the proprietary Metabolyzer™ software package. After statistical analysis and heat map generation, metabolic pathways altered 14 days after MCT injection were compared to control rats. In future (dashed arrows) this platform will be tested as a predictor for drug efficacy in the MCT and Sugen/hypoxia models of PH. Also, we will identify metabolite “fingerprints” that can be tested in clinical conditions for early PH diagnosis.