MRC Equip: A bioenergetics platform to facilitate research in metabolic pathologies
Lead Research Organisation:
Royal Veterinary College
Department Name: Comparative Biomedical Sciences CBS
Abstract
Mitochondria reside inside all cells in the body (except red blood cells), and function to generate cellular energy needed for healthy function and survival. Because they are present in all organs of the body, when they become dysfunctional, a wide variety of diseases, injuries and impairments can occur, leading to life long consequences for health and healthy ageing.
This proposal requests funding to purchase a Seahorse XF Pro Analyser. This equipment is capable of measuring tiny fluctuations in oxygen usage and pH in a variety of samples, including isolated mitochondria, cells, tissue samples and whole organisms (e.g. zebrafish larvae). This generates a snapshot of mitochondrial function, a cellular bioenergetic profile, which can then be monitored for impairments after injury or be used to report when therapeutic intervention is beneficial. The Seahorse XF Pro Analyser uses a 96-well plate format which means that many samples can be compared rapidly.
Research carried out at the RVC and elsewhere has identified mitochondrial dysfunction in the development of neurodegeneration (e.g. Parkinson's disease, Alzheimer's disease), brain injury (e.g. following oxygen deprivation during birth), cardiovascular disease, kidney disease and musculoskeletal impairment. Acquiring a Seahorse XF Pro Analyser would enable researchers to determine how target proteins implicated in these conditions perturb the cellular bioenergetic profile and how potential new therapies might restore cellular health.
This proposal requests funding to purchase a Seahorse XF Pro Analyser. This equipment is capable of measuring tiny fluctuations in oxygen usage and pH in a variety of samples, including isolated mitochondria, cells, tissue samples and whole organisms (e.g. zebrafish larvae). This generates a snapshot of mitochondrial function, a cellular bioenergetic profile, which can then be monitored for impairments after injury or be used to report when therapeutic intervention is beneficial. The Seahorse XF Pro Analyser uses a 96-well plate format which means that many samples can be compared rapidly.
Research carried out at the RVC and elsewhere has identified mitochondrial dysfunction in the development of neurodegeneration (e.g. Parkinson's disease, Alzheimer's disease), brain injury (e.g. following oxygen deprivation during birth), cardiovascular disease, kidney disease and musculoskeletal impairment. Acquiring a Seahorse XF Pro Analyser would enable researchers to determine how target proteins implicated in these conditions perturb the cellular bioenergetic profile and how potential new therapies might restore cellular health.
Technical Summary
Abnormal mitochondrial bioenergetics underpins a myriad of human diseases and identifying new therapeutic targets and treatments is vital to improving human health. To support our expanding research in the area of cellular bioenergetics, we request funding for a Seahorse XF Pro Analyser, which provides a systems-level read-out of metabolic function from cells through to zebrafish embryos and larvae.
Neurosciences and Mental Health: Bioenergetics will be performed in primary neural cells following genetic perturbation of mitochondrial dynamics, after injury or drug administration. Seahorse analyses will also be performed on cells expressing candidate risk factors for Parkinson's disease. Zebrafish models of human paediatric disease will be analysed for impaired mitochondrial function and compared with those treated with an anti-seizure drug. Zebrafish embryos and larvae will also be interrogated to determine how the impaired microbiome alters mitochondrially-regulated proteostasis, neuronal development and epilepsy.
Infection and Immunity: Bioenergetics profiling for zoonotic coccidians such as toxoplasma gondii will be trialled as a cheaper and quicker replacement for qPCR analyses. In addition, evaluating the impact of mycobacterial antigens on mitochondrial respiration will enable new control strategies to be developed for tuberculosis.
Molecular and Cellular Medicine: The Seahorse XF Pro will be used to analyse endothelial cells from humans and zebrafish models to investigate impaired angiogenesis and mechanisms of heart regeneration, as well as mitochondria-nuclear interactions in glioblastoma multiforme. Proximal tubular epithelial cells and renal cortical fibroblasts from a feline model of chronic kidney disease and macrophages from rodent models of chronic inflammatory cardiometabolic disease will be tested for metabolic impairment. Finally bioenergetics of osteocytes, chondrocytes and tenocytes in musculoskeletal injury and disease will be determined.
Neurosciences and Mental Health: Bioenergetics will be performed in primary neural cells following genetic perturbation of mitochondrial dynamics, after injury or drug administration. Seahorse analyses will also be performed on cells expressing candidate risk factors for Parkinson's disease. Zebrafish models of human paediatric disease will be analysed for impaired mitochondrial function and compared with those treated with an anti-seizure drug. Zebrafish embryos and larvae will also be interrogated to determine how the impaired microbiome alters mitochondrially-regulated proteostasis, neuronal development and epilepsy.
Infection and Immunity: Bioenergetics profiling for zoonotic coccidians such as toxoplasma gondii will be trialled as a cheaper and quicker replacement for qPCR analyses. In addition, evaluating the impact of mycobacterial antigens on mitochondrial respiration will enable new control strategies to be developed for tuberculosis.
Molecular and Cellular Medicine: The Seahorse XF Pro will be used to analyse endothelial cells from humans and zebrafish models to investigate impaired angiogenesis and mechanisms of heart regeneration, as well as mitochondria-nuclear interactions in glioblastoma multiforme. Proximal tubular epithelial cells and renal cortical fibroblasts from a feline model of chronic kidney disease and macrophages from rodent models of chronic inflammatory cardiometabolic disease will be tested for metabolic impairment. Finally bioenergetics of osteocytes, chondrocytes and tenocytes in musculoskeletal injury and disease will be determined.
