(Philadelphia, PA) - Scientists have long thought that calcium transport into mitochondria - the powerhouses of cells - is a key signal linking cardiac workload, or how hard the heart pumps, with energy production. Studies at the Lewis Katz School of Medicine at Temple University (LKSOM) and elsewhere have shown the importance of this pathway during stress, but they have also questioned the dogma that mitochondrial calcium exchange is necessary for normal cardiac function. Now, in a major breakthrough, LKSOM researchers show that the exit of calcium from mitochondria serves a critical role in heart function and may represent a powerful therapeutic approach to limit heart disease.
Using a newly developed mutant mouse model, researchers led by John W. Elrod, PhD, Assistant Professor in the Center for Translational Medicine at LKSOM, and senior investigator on the new study, demonstrate that a mitochondrial transporter encoded by the gene Slc8b1 (referred to as the mitochondrial sodium-calcium exchanger, or NCLX) is necessary for proper heart function. Without NCLX, animals suffer sudden death. The study, published online April 26 by the journal Nature, is the first to look at the necessity of mitochondrial calcium efflux in living animals.
Mitochondrial calcium exchange - the flow of calcium in and out of the energy-generating organelle - is fundamental to both cell death and pro-energetic signaling pathways. "We know from our previous work that the inhibition of calcium uptake results in a loss of stress response signaling in the heart," Dr. Elrod explained. "We found that mitochondrial calcium uptake was required for the heart to beat harder in response to stress and that excessive mitochondrial calcium uptake could trigger the death of heart cells. But those same animals had normal heart function in the absence of stress, suggesting the existence of a separate homeostatic, basal mechanism of calcium signaling."
To circumvent possible alternative mitochondrial calcium uptake pathways, Dr. Elrod and colleagues developed a conditional knockout mouse model, in which the NCLX gene was deleted after treatment with the drug tamoxifen, enabling mice to reach adulthood before the knockout was induced.
"By deleting NCLX, we were able to determine the necessity of mitochondrial calcium efflux," Dr. Elrod said.
When the gene was switched off in adult mice, the animals began to suddenly die from massive heart failure. Examination of cardiomyocytes from the animals revealed swollen and dysfunctional mitochondria, a sign of mitochondrial permeability transition pore (MPTP) activation, a mechanism known to be activated by calcium overload and to induce cell death. By genetically inhibiting MPTP activation, the researchers were able to rescue the NCLX knockout mice from death and prove the essential nature of mitochondrial calcium exchange in the heart.
Dr. Elrod and colleagues then explored the effects of augmenting NCLX expression in the mouse heart using genetic techniques. As anticipated, NCLX overexpression increased mitochondrial calcium efflux. It also prevented cell death in mice that suffered heart attacks and protected against the progression of heart failure by reducing reactive oxygen species production and limiting cardiomyocyte death and fibrosis (tissue stiffening).
"Targeting NCLX was effective in preventing cardiomyocyte death and maintaining heart function during the progression of heart failure," Dr. Elrod said. "Our findings suggest that mitochondrial calcium efflux is a promising therapeutic target, with the potential to lessen the severity of cardiac disease states."
Dr. Elrod plans to investigate NCLX activation further. Understanding its regulation at the molecular level could help identify additional mechanistic targets for the development of novel drug therapies.
Other investigators contributing to the new study include Timothy S. Luongo, a student in the Elrod lab who has recently become a postdoctoral fellow at the University of Pennsylvania; Jonathan P. Lambert, Mary Nwokedi, Alyssa A. Lombardi, Santhanam Shanmughapriya, Devin Kolmetzky, Erhe Gao, and Muniswamy Madesh, Center for Translational Medicine, Department of Pharmacology,LKSOM; Xiongwen Chen, Polina Gross, and Steven R. Houser, Center for Cardiovascular Research, Department of Physiology, LKSOM; April C. Carpenter, Department of Health and Exercise Physiology, Ursinus College; Jop H. van Berlo, Department of Medicine, University of Minnesota, Minneapolis; Emily Tsai, Division of Cardiology, Department of Medicine, Columbia University College of Physicians & Surgeons, New York; and Jeffery D. Molkentin, Department of Pediatrics, University of Cincinnati, Cincinnati Children's Hospital Medical Center, Howard Hughes Medical Institute.
The research was supported in part by National Institutes of Health grants R01 HL123966, P01 DA037830 sub-8614 and American Heart Association grants 14SDG18910041, 15PRE25080299, 16PRE31030038, and 17PRE33460423.
About Temple Health
Temple University Health System (TUHS) is a $1.6 billion academic health system dedicated to providing access to quality patient care and supporting excellence in medical education and research. The Health System consists of Temple University Hospital (TUH), ranked among the "Best Hospitals" in the region by U.S. News & World Report; TUH-Episcopal Campus; TUH-Northeastern Campus; Fox Chase Cancer Center, an NCI-designated comprehensive cancer center; Jeanes Hospital, a community-based hospital offering medical, surgical and emergency services; Temple Transport Team, a ground and air-ambulance company; and Temple Physicians, Inc., a network of community-based specialty and primary-care physician practices. TUHS is affiliated with the Lewis Katz School of Medicine at Temple University.
The Lewis Katz School of Medicine (LKSOM), established in 1901, is one of the nation's leading medical schools. Each year, the School of Medicine educates approximately 840 medical students and 140 graduate students. Based on its level of funding from the National Institutes of Health, the Katz School of Medicine is the second-highest ranked medical school in Philadelphia and the third-highest in the Commonwealth of Pennsylvania. According to U.S. News & World Report, LKSOM is among the top 10 most applied-to medical schools in the nation.
Temple Health refers to the health, education and research activities carried out by the affiliates of Temple University Health System (TUHS) and by the Katz School of Medicine. TUHS neither provides nor controls the provision of health care. All health care is provided by its member organizations or independent health care providers affiliated with TUHS member organizations. Each TUHS member organization is owned and operated pursuant to its governing documents.