New Chemical Toolkit Manipulates Mitochondria, Reveals Insights Into Drug Toxicity

FINDINGSResearchers have developed a chemical toolkit for manipulating mitochondria in its normal cellular environment. After introducing nearly 2,500 compounds to this platform—many of which are FDA-approved—the researchers immediately discerned new insights into basic mitochondrial function, which in turn revealed why some commonly used drugs have particular adverse effects.

RELEVANCEBreakdowns in mitochondria have been linked to many common diseases, such as type 2 diabetes and certain neurodegenerative conditions. In addition, mitochondria are also strongly linked to longevity. This platform provides the first comprehensive approach to probing how individual drugs affect all major aspects mitochondrial function.

INVESTIGATORVamsi Mootha, Assistant Professor of Medicine and Systems Biology, Harvard Medical School; Associate Member, Broad Institute; Center for Human Genetic Research, Massachusetts General Hospital: http://mootha.med.harvard.edu/

CITATIONNature Biotechnology, February 24, early online edition

Newswise — Why do nearly 1 million people taking cholesterol-lowering statins often experience muscle cramps? Why is it that in the rare case when a diabetic takes medication for intestinal worms, his glucose levels improve? Is there any scientific basis for the purported health effects of green tea?

A new chemical toolkit provides the first clinical explanation for these and other physiological mysteries. The answers, it turns out, all boil down to mitochondria, those tiny organelles floating around in cellular cytoplasm, often described as the cell's battery packs.

A research team led by Harvard Medical School assistant professor and Broad Institute associate member Vamsi Mootha has developed a toolkit that isolates five primary aspects of mitochondrial function and analyzes how individual drugs affect each of these areas. These results are published online February 24 in Nature Biotechnology.

Over the last few decades, mitochondria have increasingly been understood as a key determinant of cellular health. On the other hand, mitochondrial dysfunction can lead to many neurodegenerative conditions as well as metabolic diseases such as diabetes. Since mitochondria are responsible for turning the food we eat into the energy that drives our bodies, these and other connections are logical. Nevertheless, there has not yet been a systematic method for thoroughly interrogating all facets of mitochondrial activity.

"Historically, most studies on mitochondria were done by isolating them from their normal environment," says Mootha, who is also a member of the Center for Human Genetic Research at Massachusetts General Hospital. "We wanted to analyze mitochondria in the context of intact cells, which would then give us a picture of how mitochondria relate to their natural surroundings. To do this we created a screening compendium that could then be mined with computation."

In order to thoroughly analyze these organelles, Mootha and his team zeroed in on five basic features of mitochondria activity, looking at how a library of 2,500 chemical compounds affected mitochondrial toxic byproducts (like all "chemical factories" mitochondria produce their own toxic waste), energy levels, speed with which substances pass through these organelles, membrane voltage, and expression of key mitochondrial and nuclear genes. (Mitochondria contain their own genome, consisting of approximately 37 genes in humans.)

"It's just like taking your car in for an engine diagnostic," explains Mootha. "The mechanic will probe the battery, the exhaust system, the fan belt, etc., and as a result will then produce a read-out for the entire system. That's analogous to what we've done."

As a result of these investigations, Mootha and his group produced three major findings.

First, the team discovered a pathway by which the mitochondria and the cell's nuclear genome communicate with each other. They found this by discovering that certain drugs actually broke communication between these two genomes. By reverse engineering the drugs' toxic effects, they may be able to reconstruct normal function.

Second, the team looked at a class of the cholesterol-lowering drugs called statins. Roughly 100 million Americans take statins, and among that group, about 1 million experience muscle cramping and aches. Previous studies suggested that mitochondria were involved, but clinical evidence remained conflicting. Mootha and his colleagues found that three out of the six statins (Fluvastatin, Lovastatin, and Simvastatin) interfered with mitochondria energy levels, as did the blood-pressure drug Propranolol. When combined, the effect was worse.

"It's likely that a fair number of patients with heart disease are on one of these three statins as well as Propranolol," says Mootha, "Our cellular studies predict that these patients might be at a higher risk for developing the muscle cramps. Obviously, this is only a hypothesis, but now this is easily testable."

The third and arguably most clinically relevant finding builds on a paper Mootha coauthored in 2003, a paper that demonstrated how type 2 diabetes was linked to a decrease in the expression of mitochondrial genes. A subsequent and unrelated paper showed a relationship between type 2 diabetes and an increase in mitochondrial toxic byproducts. Mootha's group decided to query their toolkit and see if there were any drugs that affected both of these functions, drugs that could boost gene expression while reducing mitochondrial waste.

Indeed, they found six compounds that did just that, five of which were known to perturb the cell's cytoskeleton, that is, the scaffolding that gives a cell its structure.

"Our data shows that when we disrupt the cytoskeleton of the cell, that sends a message to boost the mitochondria, turning on gene expression and dropping the toxic byproducts," says Mootha. "The connection between the cytoskeleton and mitochondrial gene expression has never been shown before and could be very important to basic cell biology."

Of the five drugs that did this, one, called Deoxysappanone, is found in green tea and is known to have anti-diabetic effects. Another, called Mebendazole, is used for treating intestinal worm infections. This connection gives a rationale to case reports in which diabetics treated with Mebendazole have described improvements in their glucose levels while on the drug.

The researchers intend to further investigate some of the basic biological questions that this study has raised, foremost being the relationship between the cytoskeleton and mitochondria. They also plan on using this toolkit to develop strategies for restoring normal mitochondrial function in certain metabolic and neurodegenerative conditions where it has broken down.

This research was funded by grants from the American Diabetes Association and the Richard and Susan Smith Family Foundation.

Full citation:

"Large-scale chemical dissection of mitochondrial function"

Bridget K Wagner(1,5), Toshimori Kitami1(2,5), Tamara J Gilbert(1), David Peck(1), Arvind Ramanathan(1), Stuart L Schreiber(1), Todd R Golub(1,3) & Vamsi K Mootha(1,2,4)

1-Broad Institute of Massachusetts Institute of Technology and Harvard, Seven Cambridge Center, Cambridge, MA2-Center for Human Genetic Research, Massachusetts General Hospital, 185 Cambridge Street, Boston, MA3-Department of Pediatric Oncology, Dana-Farber Cancer Institute, 44 Binney Street, Boston, MA4-Department of Systems Biology, Harvard Medical School, 200 Longwood Avenue, Boston, MA5-These authors contributed equally to this work.

Harvard Medical School (http://hms.harvard.edu/hms/home.asp) has more than 7,500 full-time faculty working in 11 academic departments located at the School's Boston campus or in one of 47 hospital-based clinical departments at 17 Harvard-affiliated teaching hospitals and research institutes. Those affiliates include Beth Israel Deaconess Medical Center, Brigham and Women's Hospital, Cambridge Health Alliance, Children's Hospital Boston, Dana-Farber Cancer Institute, Forsyth Institute, Harvard Pilgrim Health Care, Joslin Diabetes Center, Judge Baker Children's Center, Immune Disease Institute, Massachusetts Eye and Ear Infirmary, Massachusetts General Hospital, McLean Hospital, Mount Auburn Hospital, Schepens Eye Research Institute, Spaulding Rehabilitation Hospital, and VA Boston Healthcare System.

The Broad Institute's (http://www.broad.mit.edu/index.html) mission is to fulfill this promise by empowering creative scientists to construct new powerful tools for genomic medicine, to make them accessible to the global scientific community, and to apply them to the understanding and treatment of disease. The Institute is a research collaboration involving faculty, professional staff and students from throughout the MIT and Harvard academic and medical communities and is governed jointly by the two universities. It was founded in 2003 through the far-sighted generosity of philanthropists Eli and Edythe Broad. The Institute is organized around Scientific Programs and Scientific Platforms. This unique structure enables scientists to collaborate on important projects, with the potential to transform medicine.

Massachusetts General Hospital (http://www.massgeneral.org), established in 1811, is the original and largest teaching hospital of Harvard Medical School. The MGH conducts the largest hospital-based research program in the United States, with an annual research budget of more than $500 million and major research centers in AIDS, cardiovascular research, cancer, computational and integrative biology, cutaneous biology, human genetics, medical imaging, neurodegenerative disorders, regenerative medicine, systems biology, transplantation biology and photomedicine.