Oxygen consumption rate provides a quantitative assessment of how hard the cell has to work to meet its energetic and biosynthetic demands. This application enables investigators to determine the level of mitochondrial activity necessary to enable and regulate cellular functions such as proliferation, activation, differentiation, migration and death.

Metabolic switching is how cells reprogram bioenergetics in order to achieve different phenotypes. Often this is due to a change in nutrients and/or metabolic pathways such as mitochondrial respiration and glycolysis. This application enables investigators to measure and monitor mitochondrial activity over days and even weeks. By continuously tracking oxygen consumption rate the contribution of mitochondria to switches resulting in changes in cellular function can be identified, followed and even controlled.

Given the fundamental role of mitochondria in the viability and vitality of cells, impairment of this organelle is a universal cause and/or consequence of cellular dysfunction, impairment, and death. It is often the cause of many resulting diseases. This application allows you to non-invasively measure and track mitochondrial respiration in any standard cell culture model system, providing a democratized, objective measure of cell function. The ability to standardize levels of impairment and function that initiate and/or maintain disease pathologies, whether acute or chronic, would improve the comparability of data from other analytical methods.

Stem-cell derived models have opened up entirely new and more predictive ways of modeling disease leading to new discoveries, translatable findings and validation of interventional strategies. This application continuously monitors mitochondrial activity in a label free and non-invasive manner. This enables objective control of the differentiation process, free of the potential bias and complexity of biomarkers and other invasive techniques.

The FDA's recognition of the need for more predictive human cell models in preclinical testing provides credence to advances in the ability to generate, monitor and quality control human preclinical models of disease. Resipher oxygen consumption rate provides objective, label free, continuous measures of mitochondrial activity. This improves consistency and reliability of preclinical disease models due to standardization and quality control.

It is now appreciated that the ability to perform more productive and predictive in vitro cell assays requires more control and monitoring of cell culture conditions. Mitochondrial respiration has been shown to provide an objective and actionable measure of in vitro cell behavior that can be 'tuned' for different cell model systems. The Resipher oxygen consumption rate platform is the only system capable of continuous, multi-day, label free measure of OCR that is agnostic to culture plate, media and conditions.

In order to enable in vitro cell modeling to be more relevant and predictive, the environmental conditions the cells are experiencing need to better reflect in vivo conditions. Mitochondria react to and integrate many of the external stimuli and environmental conditions, such as oxygen, that affect cellular behavior. Oxygen levels in cell culture rarely reflect oxygen availability in vivo (i.e. normoxia). As well, diseases like cancer and infection create altered oxygen conditions (most often hypoxia) that are adverse to the host. Using the Resipher system to measure and monitor oxygen consumption rates can determine if oxygen conditions are appropriate and/or is confounding mitochondrial function.

Mitochondrial bioenergetic pathways have become increasingly targeted beyond rare metabolic disorders into age-related diseases with high unmet need such as neurodegeneration, obesity/diabetes, cardiomyopathies and most recently immuno-oncology. Drug strategies that inhibit or increase mitochondrial function, especially in chronic disease, must show persistence and low toxicity over significant periods of time. This application will detect changes in metabolic function due to either direct targeting of the mitochondria or a consequence of upstream drug activity. Because the Resipher system can continuously measure alterations in mitochondrial activity over days and weeks it is more likely to detect meaningful changes in drug activity and/or adverse effects in chronic disease models at relevant doses.

Whether directly targeting mitochondrial pathways or not, that ability of a drug to mediate its activity will most likely alter the cellular metabolic programming detectable by changes in oxygen consumption rate. Dose-dependent, drug response assays that are performed over several hours to a few days are often not sufficient to predict low dose and/or chronic affects which require more time to manifest, both therapeutic and adverse effects. This application will identify and measure changes in mitochondrial activity in a dose dependent manner. This can be correlated with therapeutic activity and/or potential adverse effects. The ability to measure continuously over long periods of time increases the chance to observe activity at lower doses as well as drug effects that take longer to manifest in relevant cell models.

The ability to deliver along with the regulatory push for more predictive, human in vitro preclinical models, has led to an explosion in 3D and organ-on-a-chip culture systems. Because these higher-order, functioning biological systems take weeks and even months to achieve the appropriate level of differentiation and integrated function it necessitates non-invasive, continuous monitoring to control and determine when the appropriate functionality has been acquired. This application constantly monitors mitochondrial activity and switching in order to ensure the modeling behavior of the cell culture is appropriate and consistent.

Key CQA's like persistence, engraftment, proliferative potential, renewal, etc. are qualities that make 'living' drugs uniquely potent and durable. Conventional biomarkers and molecular tools are not able to capture the dynamic and heterogenic nature of this class of drugs. In this application Resipher can measure objective changes in mitochondrial activity that can be correlated with specific cell phenotypes and function. Three well-established examples are:
— Increases in mitochondrial function that enable memory T cell function
— Metabolic switching regulating programmatic differentiation of neural precursor into differentiated functioning circuits
— Measuring mitochondrial dysfunction when it is reversible, i.e. before the emergence of disease pathologies.

This has been especially challenging as living cells have more dynamic behavior and complex composition. Because metabolism must be in equilibrium, measurable metabolic features such as oxygen consumption rates, can be stable indicators of cell quality, functional capacity and potency. This application can be used to establish defined levels of mitochondrial respiration that are indicative of therapeutic activity, persistence, safety, potency, etc.

The defined composition and highly predictive nature of small molecule drugs and more recently protein biologics have lent themselves to 'recipe' style, batch processing. However, the starting point and determination of potency for cell therapies will require additional and different tools to account for their heterogeneous composition and dynamic equilibrium. This application can be used to identify the appropriate level and/or range of mitochondrial activity of the product based on multiple factors such as cell type, media, engineering, etc. Once this is established then the bioreactor can be monitored to induce and/or control cell manufacturing to reach the desired cell therapy dose threshold or range.