Expert view: Following mitchondrial substrate oxidation pathways to expose novel drug targets

Such discoveries have been facilitated in no small part by the development of specific empowering technologies, including the Agilent Seahorse XF platform. By making in vitro metabolic assessment more accessible, these real-time live-cell metabolic assays have helped reveal the pivotal role of metabolic reprogramming in disease progression. What these assessments are now showing is that knowledge of which specific metabolic substrates drive particular processes provides a much deeper understanding of how cell metabolism and cell function intersect. This, in turn, offers important opportunities for drug discovery, whereby metabolic modulations become therapeutic targets or as means to drive immune cells to specific fates.

To further enable these investigations, a suite of optimised assays for measuring cellular substrate oxidation has been developed for the XF platform. These Substrate Oxidation Stress Tests interrogate three primary mitochondrial substrates: long chain fatty acids, glucose/pyruvate and/or glutamine by assessing cellular sensitivity to specific pathway inhibitors. As measurements are also performed under conditions of elevated substrate oxidation, the tests
have the ability to reveal inherent or specific substrate demands that are often not apparent under basal conditions but can become immensely important from a translational perspective.

These methods therefore enable a deeper understanding of specific substrate oxidation processes, their impact on fundamental cellular functions and the ability to characterise responses to genetic manipulations, pharmaceutical interventions or disease-relevant microenvironments. For more information, visit www.agilent.com/chem/discoverXF.