Discovery of mitochondrial protein by researchers at Lewis Katz School of Medicine at Temple University opens path to therapeutic advances for heart and Alzheimer’s disease

(Press-News.org) (Philadelphia, PA) – Calcium transport into and out of mitochondria – the powerhouses of cells – is central to cellular energy production and cell death. To maintain the balance of calcium within these powerhouses, cells rely on a protein known as the mitochondrial sodium-calcium exchanger, or NCLX. Now, in new research, scientists at the Lewis Katz School of Medicine at Temple University have discovered a novel regulator of NCLX activity, a protein called TMEM65, which helps move calcium out of mitochondria, protecting against harmful calcium overload.

The discovery, described online April 8 in the journal Nature Metabolism, is the first to characterize the interaction of TMEM65 with NCLX in mitochondria. “TMEM65 is the first protein identified that is a bona fide interactor and regulator of NCLX,” explained John W. Elrod, PhD, W.W. Smith Chair in Cardiovascular Medicine and Founding Director of the Aging + Cardiovascular Discovery Center at the Lewis Katz School of Medicine and senior investigator on the new study. The discovery could help scientists design new therapeutic agents to combat calcium overload of mitochondria in conditions such as heart failure and Alzheimer’s disease.

Mitochondrial calcium exchange serves a critical part in regulating cell survival and pro-energetic signaling pathways. When mitochondria take in too much calcium, which can happen in certain disease states, energy metabolism is disrupted and cells die. This is most apparent in the heart, where calcium overload contributes to the permanent loss of heart muscle cells during heart attacks and in heart failure. It also can result in the loss of brain cells in Alzheimer’s disease and other neurodegenerative conditions.

Dr. Elrod and colleagues previously identified NCLX as a key player in the removal of calcium from mitochondria in the heart and brain. Research has also shown that augmenting NCLX activity can limit the progression of not only heart failure and Alzheimer’s disease but also cancer. Nonetheless, despite these promising findings, an understanding of the mechanisms underlying NCLX regulation has remained elusive.

“NCLX has a very complex structure, which has impeded the study of its regulation and hindered progress in therapeutic development,” Dr. Elrod said. “For our latest study, we decided to take a different approach, using biotin tagging, which allowed us to trace NCLX’s interactions with other proteins in intact cells.”

Led by postdoctoral fellow Joanne F. Garbincius, PhD, Dr. Elrod’s team generated a fusion of NCLX and a biotinylation protein. The fusion protein was then placed back into cells, and other proteins that came within its proximity were biotinylated, or biochemically labeled. The biotinylated molecules were then easily isolated, enabling their identification with mass spectrometry. In this way, the researchers ultimately discovered TMEM65 as a primary suspect in NCLX regulation.

“TMEM65 was of particular interest because it is a mitochondrial protein of unknown function,” Dr. Elrod explained. “We also knew about a case report in which a young girl with a loss-of-function mutation in TMEM65 experienced profound muscle weakness and microcephaly (abnormally small head/brain) and neurological dysfunction.”

In subsequent experiments, it was discovered that when TMEM65 is removed from cells, calcium levels in the mitochondria accumulate. This led to the realization that TMEM65 is required for NCLX activity. Its role in regulating NCLX was confirmed in a mouse model in which TMEM65 levels were significantly decreased. As animals matured, they experienced a progressive loss of neuromuscular function, to the extent that they could barely walk by adulthood.

The methods used to identify TMEM65 and to elucidate NCLX regulation are groundbreaking in the field of basic cardiovascular science. In 2024, Dr. Garbincius was recognized for her research with the prestigious American Heart Association’s Louis N. and Arnold M. Katz Basic Science Research Prize for Early Career Investigators.

The work has also inspired ongoing investigation of TMEM65. Dr. Elrod and colleagues plan next to explore the possibility of modulating TMEM65 activity as a therapeutic strategy. “TMEM65 is a promising therapeutic target,” Dr. Elrod added. “Figuring out how to augment or otherwise alter its interaction with NCLX could offer an important treatment option for patients affected by diseases involving pathogenic calcium buildup in mitochondria.”

Amy J. Goldberg, MD, FACS, The Marjorie Joy Katz Dean of the Lewis Katz School of Medicine, emphasized the significance of this research, stating, “This discovery exemplifies the transformative science happening at the Lewis Katz School of Medicine. By deepening our understanding of mitochondrial function, our researchers are paving the way for innovative treatments that could have a profound impact on patients with heart failure, Alzheimer’s disease, and beyond.”

Other researchers involved in the study include Oniel Salik, Henry M. Cohen, Carmen Choya-Foces, Adam S. Mangold, Angelina D. Makhoul, Anna E. Schmidt, Dima Y. Khalil, Joshua J. Doolittle, Anya S. Wilkinson, Emma K. Murray, Michael P. Lazaropoulos, and Alycia N. Hildebrand, Aging + Cardiovascular Discovery Center, Department of Cardiovascular Sciences at the Lewis Katz School of Medicine; and Dhanendra Tomar, Department of Internal Medicine, Wake Forest University School of Medicine, Winston-Salem, North Carolina.

The research was supported in part by funding from the National Institutes of Health and the American Heart Association.

About the Lewis Katz School of Medicine

Founded in 1901, the Lewis Katz School of Medicine at Temple University attracts students and faculty committed to advancing individual and population health through culturally competent patient care, research, education, and service. The School confers the MD degree; MS and PhD degrees in Biomedical Science; the MA in Urban Bioethics; the MS in Physician Assistant studies; a certificate in Narrative Medicine; a non-degree post-baccalaureate program; several dual degree programs with other Temple University schools; continuing medical education programs; and in partnership with Temple University Hospital, 40 residency and fellowship programs for physicians. The School also manages a robust portfolio of publicly and privately funded transdisciplinary studies aimed at advancing the prevention, diagnosis, and treatment of disease -- with specialized research centers focused on heart disease, cancer, substance use disorder, metabolic disease, and other regional and national health priorities. To learn more about the Lewis Katz School of Medicine, please visit: medicine.temple.edu.

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