Cellular energy metabolism underlies diverse, essential functions including cell division, differentiation, and the ability to carry out effector functions. Cell metabolic states, and more importantly, metabolic switches, are activated not only in response to oxygen (O2) deprivation or nutrient availability/starvation, but also in re- sponse to infection and tissue damage. In mammalian cells, the two major pathways that produce energy are glycolysis and oxidative phosphorylation. The Agilent Seahorse XFe96 Analyzer measures key metabolic pa- rameters, pH change and O2 consumption, byproducts of glycolysis and oxidative phosphorylation, respective- ly. This instrument carries out these measurements with high sensitivity in real time, allowing for testing under diverse conditions (e.g., various stimuli, multiple pathway-specific inhibitors, various substrates) simultaneously and with high replicate numbers (due to a 96-well format), ultimately contributing to improved scientific rigor. The investigators assembled in this revised proposal are brought together by a need for access to Seahorse extracellular flux technology that cannot be addressed through the existing systems at the University of Mary- land School of Medicine (UMSOM). Each of their diverse Research Projects has a distinct need addressable by the Seahorse XFe96 Analyzer. These include analyses of: metabolic changes associated with macrophage differentiation during infection (Vogel, PI); mechanisms by which Rickettsiae co-opt host metabolic pathways for their own survival (Azad); metabolic dysregulation underlying dendritic cell immunogenicity (Cao); stimula- tion of tendon repair by metabolic modifiers (Enomoto-Iwamoto); neurotoxicity/neuroinflammation in injured brain (Fiskum, Loane, Polster, Stoica); role of oxidative phosphorylation in tumor suppression (Kalvakolanu); bioenergetics and inflammation as mechanisms for post-stroke fatigue (Klinedinst); regulation of O2 consump- tion and ATP production by mitochondrial matrix calcium (Lederer); reduction of Complex I to V subunits in mi- tochondrial oxidative phosphorylation in neurodegeneration (Monteiro); control of inflammation and metabolic aberrations in obesity (Moudgil); metabolic changes resulting in RUNX2-mediated transcriptional regulation of tumor growth (Passaniti); glucose-induced metabolic reprogramming of premalignant keratinocytes (Schnei- der); role of pyruvate dehydrogenase mutations in oxidative metabolism of glucose in obesity (Taylor, Mitchell); mechanisms by which maternal hypoxia disrupts trophoblast invasion of spiral arteries in the placenta (Thomp- son); the role of miR-140 in cancer stem cell transformation and the impact of Nrf2 activation on mammary epi- thelial cell metabolism (Zhou). This instrument will be housed in the UMSOM Center for Innovative Biomedical Research (CIBR) centralized core and will support not only Users named in this application, but also, provide broad access to all UMSOM investigators. Coupled with technical experts and administrative support, and a significant level of institutional commitment, we anticipate that the Seahorse XFe96 Analyzer will accelerate development of novel therapies that target metabolic dysregulation associated with multiple disease states.
The Seahorse XFe 96 Analyzer measures changes in cellular metabolism and is anticipated to have a major impact on research into new therapies and treatments for a number of complex diseases including infection, cancer, traumatic brain injury, diabetes, obesity, and neurodegenerative disease.
