Consuming a high-fat diet can lead to a variety of health problems — not only weight gain but also an increased risk of diabetes and other chronic diseases.At the cellular level, hundreds of changes take place in response to a high-fat diet. MIT researchers have now mapped out some of those changes, with a focus on metabolic enzyme dysregulation that is associated with weight gain.Their study, conducted in mice, revealed that hundreds of enzymes involved in sugar, lipid, and protein metabolism are affected by a high-fat diet, and that these disruptions lead to an increase in insulin resistance and an accumulation of damaging molecules called reactive oxygen species. These effects were more pronounced in males than females.The researchers also showed that most of the damage could be reversed by giving the mice an antioxidant along with their high-fat diet.“Under metabolic stress conditions, enzymes can be affected to produce a more harmful state than what was initially there,” says Tigist Tamir, a former MIT postdoc. “Then what we’ve shown with the antioxidant study is that you can bring them to a different state that is less dysfunctional.”Tamir, who is now an assistant professor of biochemistry and biophysics at the University of North Carolina at Chapel Hill School of Medicine, is the lead author of the new study, which appears today in Molecular Cell. Forest White, the Ned C. and Janet C. Rice Professor of Biological Engineering and a member of the Koch Institute for Integrative Cancer Research at MIT, is the senior author of the paper.Metabolic networksIn previous work, White’s lab has found that a high-fat diet stimulates cells to turn on many of the same signaling pathways that are linked to chronic stress. In the new study, the researchers wanted to explore the role of enzyme phosphorylation in those responses.Phosphorylation, or the addition of a phosphate group, can turn enzyme activity on or off. This process, which is controlled by enzymes called kinases, gives cells a way to quickly respond to environmental conditions by fine-tuning the activity of existing enzymes within the cell.Many enzymes involved in metabolism — the conversion of food into the building blocks of key molecules such as proteins, lipids, and nucleic acids — are known to undergo phosphorylation.The researchers began by analyzing databases of human enzymes that can be phosphorylated, focusing on enzymes involved in metabolism. They found that many of the metabolic enzymes that undergo phosphorylation belong to a class called oxidoreductases, which transfer electrons from one molecule to another. Such enzymes are key to metabolic reactions such as glycolysis — the breakdown of glucose into a smaller molecule known as pyruvate.Among the hundreds of enzymes the researchers identified are IDH1, which is involved in breaking down sugar to generate energy, and AKR1C1, which is required for metabolizing fatty acids. The researchers also found that many phosphorylated enzymes are important for the management of reactive oxygen species, which are necessary for many cell functions but can be harmful if too many of them accumulate in a cell.Phosphorylation of these enzymes can lead them to become either more or less active, as they work together to respond to the intake of food. Most of the metabolic enzymes identified in this study are phosphorylated on sites found in regions of the enzyme that are important for binding to the molecules that they act upon or for forming dimers — pairs of proteins that join together to form a functional enzyme.“Tigist’s work has really shown categorically the importance of phosphorylation in controlling the flux through metabolic networks. It’s fundamental knowledge that emerges from this systemic study that she’s done, and it’s something that is not classically captured in the biochemistry textbooks,” White says.Out of balanceTo explore these effects in an animal model, the researchers compared two groups of mice, one that received a high-fat diet and one that consumed a normal diet. They found that overall, phosphorylation of metabolic enzymes led to a dysfunctional state in which cells were in redox imbalance, meaning that their cells were producing more reactive oxygen species than they could neutralize. These mice also became overweight and developed insulin resistance.“In the context of continued high fat diet, what we see is a gradual drift away from redox homeostasis towards a more disease-like setting,” White says.These effects were much more pronounced in male mice than female mice. Female mice were better able to compensate for the high fat diet by activating pathways involved in processing fat and metabolizing it for other uses, the researchers found.“One of the things we learned is that the overall systemic effect of these phosphorylation events led to, especially in males, an increased imbalance in redox homeostasis. They were expressing a lot more stress and a lot more of the metabolic dysfunction phenotype compared to females,” Tamir says.The researchers also found that if they gave mice who were on a high-fat diet an antioxidant called BHA, many of these effects were reversed. These mice showed a significant decrease in weight gain and did not become prediabetic, unlike the other mice fed a high-fat diet.It appears that the antioxidant treatment leads cells back into a more balanced state, with fewer reactive oxygen species, the researchers say. Additionally, metabolic enzymes showed a systemic rewiring and changed state of phosphorylation in those mice.“They’re experiencing a lot of metabolic dysfunction, but if you co-administer something that counters that, then they have enough reserve to maintain some sort of normalcy,” Tamir says. “The study suggests that there is something biochemically happening in cells to bring them to a different state — not a normal state, just a different state in which now, at the tissue and organism levels, the mice are healthier.”In her new lab at the University of North Carolina, Tamir now plans to further explore whether antioxidant treatment could be an effective way to prevent or treat obesity-associated metabolic dysfunction, and what the optimal timing of such a treatment would be.The research was funded in part by the Burroughs Wellcome Fund, the National Cancer Institute, the National Institutes of Health, the Ludwig Center at MIT, and the MIT Center for Precision Cancer Medicine.
