Trust is Good, Control is Better
(April 17th, 2015) US and European scientists recently dropped a bombshell on research involving the miracle fat-burning hormone, irisin. Their case is a perfect example for demonstrating the overreliance on commercially available test kits.
Scientists are humans, and as such, they can sometimes get carried away when they make a breakthrough discovery. Because of this premature excitement they may lose attention to detail, over-interpret results, or cut corners to speed up that much-desired Nature publication. The discovery of irisin, or ‘exercise hormone’, is one such example. Once thought to be a promising exercise-free solution to fight obesity and diabetes, irisin has now been shown to be not more than a random blood protein detected by flawed reagents.
Bruce Spiegelman and colleagues at Harvard Medical School (US) first discovered irisin in 2012. In a Nature article the researchers reported that after exercise, muscle cells release a fragment of a pre-hormone-like protein called FNDC5 into the bloodstream where it travels to adipose cells to trigger the conversion of white fat into calorie-burning brown fat. They concluded that this small molecule is a “newly identified hormone” which they named irisin, after the Greek messenger goddess Iris.
Whereas white fat stores energy brown fat is converted to heat - that’s how hibernating animals and newborn babies stay warm. So, unless you starve or exercise a lot, your white fat will remain stubbornly lodged on your hips while brown fat burns calories. Unfortunately for most of us though, only about 10% of our adipose tissue consists of brown fat-producing cells. And this is why the discovery of irisin was so exciting. What if we could take an irisin pill to turn our white fat into brown fat? Could we burn calories while lying comfortably on the couch eating ice cream?
It is no surprise then that in just three years over 170 studies were published on irisin. It didn’t take that long for someone to question the Spiegelman study, though. Harold Erickson from Duke University (US) first voiced his concerns about irisin in 2013 and recently, he and an international team of four research groups clearly showed that the most widely used, commercial antibodies against irisin are unspecific - they detect cross-reaction blood proteins, basically unknown random proteins.
In the original irisin paper Spiegelman’s team identified irisin with a polyclonal antibody produced by Abcam that should in theory attach to the tail of FNDC5. But irisin is a fragment of FNDC5 that is chopped from the other end of the protein, so this antibody couldn’t possibly detect it, Erickson argued. Spiegelman replied to this by saying that Abcam had not correctly annotated the antibody in their catalogue.
Erickson also noticed that none of the commercially available irisin antibodies had been properly tested by the companies that had made them. But despite this worrying observation, several research groups continued to use them, and what’s worse, without attempting to verify their specificity for irisin. So, who’s really to blame? The companies selling untested products or the researchers blindly relying on them?
That’s a difficult question. What’s certain, though, is that at the moment there seems to be no trustable irisin antibody on the market. In their latest study Erickson and colleagues tested four commercial irisin antibodies used in over 80 studies by Western Blotting. To be sure they were looking at the right thing the researchers synthesised irisin molecules and then compared them side-by-side with the proteins detected by the commercial antibodies. They tested several tissue samples from humans and other animals, including blood serum from horses after strenuous exercise. None of the antibodies detected a protein with the size of irisin in any of the samples, and even more worrying, they didn’t detect synthesised irisin. However, the antibodies reacted with many other proteins of the wrong size. This shows that all previously published studies based on assays using these antibodies “were reporting unknown cross-reacting proteins”, the authors claim in the study.
So, does irisin exist at all? To answer this question, the team looked for irisin in human blood serum using mass spectrometry. They were able to identify a molecule corresponding to FNDC5 or irisin, which is the “first mass spectrometry identification of an irisin peptide at the correct size, and might be considered as supporting the existence of irisin in human serum,” the authors say in the study. However, the very low amounts of irisin detected “makes a physiological role for irisin very unlikely”. According to Erickson and colleagues, the exercise hormone is a myth.
These new findings are bad news for irisin researchers and food lovers, but they’re very good news for science. They show that even though human nature might at times corrupt scientific discoveries (voluntarily or involuntarily), science infallibly corrects itself. Hence, although we can’t trust every commercial antibody, we can, at least, trust the scientific process.