Spit It Out! How saliva may provide the key to sports-related concussion

concussionSRxA’s Word on Health has reported several times on the problems of concussion among football players and cheerleaders. So we couldn’t help but sit up and take notice of an article in our local newspaper about some cutting edge research taking place yards from our front door.

The third floor of Bull Run Hall on George Mason University’s Prince William campus plays host to plenty of mind-bending science projects – laser capture microdissection, protein electronics and high-resolution mass spectrometry to name but a few.

Yet the most complex and potentially influential findings to emerge from the school’s College of Science might come down to a kid spitting in a cup after football practice.

Once a week athletic trainers collect saliva samples from the 12- and 13-year-old kids playing for the Jets, an A-League football team in the Central Loudoun Youth Football League. Then they send them to Dr. Shane Caswell, a George Mason professor and pioneer of the world’s first salivary biobank designed for concussion research in athletes.

Caswell stores the saliva in a freezer he dubs the “spit repository.” He eventually extracts the samples and runs them through sophisticated machinery to determine changes in protein variance. By comparing each kid’s spit samples to previous submissions, he hopes to uncover a handful of proteins that can detect concussions.

George Mason scientistsWorking alongside Caswell is Dr. Chip Petricoin. Long accustomed to studying protein biomarkers for cancer research, Petricoin never imagined he’d wind up plying his trade for studies on traumatic brain injury and concussions.  But the seed was planted six years ago, when he was called up to Fort Detrick to conduct a site review for a company that had been given a grant from the army to do concussion research. Petricoin admired their efforts, but he realized that his own work with cancer biomarkers could reap significant benefits for the concussion research that remained in its nascent stages.

A year later he found himself working in the same building as Caswell, whose extensive background in athletic training was getting him increasingly involved with concussion research. The two discussed their respective endeavors and quickly realized they could join forces.

The university’s College of Science and College of Education and Human Development began funding their efforts last year. Since then, Caswell and Petricoin have begun to explore the vast quantities of information stored in athletes’ salivary biomarkers.

Think about the biomarker content of a sample like an iceberg,” said Petricoin, co-director of the university’s Center for Applied Proteomics and Molecular Medicine. “The concept of what you see is only the tip of the iceberg? That’s kind of like biomarker research. Things that have been seen before are just the tip of what really is there. If we could go all the way down and see everything, you’d see a whole new iceberg. So we’re kind of going a mile deep now in the iceberg biomarker research.”

athlete spittingIndeed, the depth of these largely uncharted waters became apparent when the duo began their work with the Jets this fall using eight saliva samples. A few weeks after collecting those baseline samples, they used the nanotechnology at their disposal to examine new samples from four of the same kids who had recently suffered concussions. After compiling a list of proteins, they found that 60% of their list featured proteins that had never been described.

The process generates an information archive that’s larger than anyone’s ever seen before in saliva,” Petricoin said.

Caswell and Petricoin are currently working on 37 concussion cases, a total that increases every week with new samples arriving from different sources. They’re collaborating with Prince William County Public Schools, as well as intercollegiate athletics at Marymount University and George Mason.

The Jets’ head coach, Rob Scola, says his team has so far adapted nicely to the study. George Mason sends a certified athletic trainer to the field to provide care and to collect data on hits the players endure. The trainer tapes every game and practice, something that allows coaches to see what they’re doing right and wrong in their efforts to teach proper heads-up tackling techniques. Players also wear helmets with sensors that detect the force and location of impacts sustained in practices and games.

It’s all part of an effort to determine what measures coaches should take to minimize players’ risk of head trauma on the football field, where the rate of brain injuries is higher than in any other youth sport.

It’s very hard to get information from a very small team in a very small league and then extrapolate that,” Scola said. “I think that as Mason starts to expand the study, I think there will be some really interesting pieces of information that come from that, which I believe can be helpful to the league and football as a whole. I think it’s a phenomenal first step.”

Part of the project’s appeal lies in its lack of hassle. Biomarker work has traditionally come from blood and spinal fluid samples, which are rooted in far more invasive processes than simply spitting in a cup.

If I were to go out on the field and say, ‘Hold on a second. I want to take your child’s blood or their cerebral spinal fluid.’ That’s game over. We can’t move forward,” Caswell said. “This is a non-invasive tool that is rapidly deployable. There’s no threat of infection, it’s easily done and it provides a great deal of information.”

Caswell and Petricoin’s work with the Jets has opened the door to broader studies that extend to the entire lifespan of an athlete’s career. The hope is that parents will have their children give samples when they begin participating in youth football, ice hockey, soccer, or whatever sport they choose to play. They can then follow that up by giving more samples as they pursue the sport in high school, college and beyond.

“You are then able to track at various time points throughout someone’s career and identify how their marker is changing and then maybe one day compare it to a database that could help inform decisions about whether or not that individual should retire from play, whether that individual is suffering any adverse consequences from their participation,” Caswell said.

blue mouthgardEven more ambitious is their ultimate goal of implementing the biomarkers into a clinical diagnostic device. Petricoin envisions a mouthguard that turns from clear to blue when a concussion is detected. The technology, he says, is there – impregnating the nanoparticles into the mouth guard, binding the biomarkers and producing a color shift are concepts that have already been engineered.

The hard part is nailing down the biomarkers. For the moment, all Caswell and Petricoin are trying to do is identify what’s in the saliva. As the data mounts, they hope to reveal protein distribution patterns that coincide with repeated head trauma.

Caswell, a former hockey player who once returned to the ice minutes after suffering a concussion only to realize minutes later that he wasn’t carrying his stick, believes those patterns will come and that his team is well-positioned to make meaningful discoveries that could impact concussion policies on a greater scale.

If and when they do, we’ll be sure to let you know.

SRxA-logo for web

Diabetes Drug may Repair Injured Brains

Here’s a good brain teaser for a Wednesday.  What do an old diabetes drug, brain injury and Alzheimer’s Disease have in common?

Here’s some clues to help you solve the riddle.

(i)           Metformin is a widely used treatment for type II diabetes

(ii)          An increasing proportion of people with Alzheimer’s Disease also have diabetes

(iii)         Hyperinsulinemia (excess levels of insulin in the blood) may enhance the onset and progression of neurodegeneration

Have you solved it?  If so, congratulations!

If not, the answer, according to data just published in the journal Cell Stem Cell is that the former may hold the clue to treating the latter.

In other words, the study suggests that metformin, an anti-diabetes drug first discovered in the 1920’s, is able to help activate the mechanism that signals stem cells to generate brain cells.

Principal investigator, Freda Miller, a Professor from the Department of Molecular Genetics at the University of Toronto
says “If you could take stem cells that normally reside in our brains and somehow use drugs to recruit them into becoming appropriate neural cell types, then you may be able to promote repair and recovery in at least some of the many brain disorders and injuries for which we currently have no treatment.”

The research involved laboratory experiments using both mouse and human brain stem cells, as well as learning and memory tests performed on live mice given the drug.

Miller and her colleagues started by adding metformin to stem cells from the brains of mice, then repeated the experiment with human brain stem cells generated in the lab. In both cases, the stem cells gave rise to new brain cells.

They then tested the drug in lab mice and found that those given daily doses of metformin for two or three weeks had increased brain cell growth and outperformed rodents not given the drug in learning and memory tasks.

In the key experiment, mice were forced to learn the position of a platform hidden under the surface of a water-filled maze and then asked rapidly to learn a new position.

Mice were injected with either metformin or saline for 38 days. On days 22 through 38, they learned the initial position of the platform, which provided an escape from the water-filled maze.  Then the platform was moved to the opposite side of the maze, and the animals were asked again to learn its position. In both tasks, the mice learned the platform positions with equivalent speed.

But when they were put back in the maze – this time with the platform removed – control mice spent more time searching for it in the original position, while the metformin-treated animals preferentially looked in the new region.

The implication  is that metformin helped the mice form their new memories of the second platform position. Further analysis showed that their enhanced ability was paralleled by an increase in the number of  neurons.

In a separate study researchers have shown that metformin can increase lifespan and delay the onset of cognitive impairment in a mouse model of Huntington’s disease.

Taken together, these findings raise the possibility that metformin’s ability to enhance neurogenesis might have a positive impact in some nervous system disorders.

Miller’s team is already planning a pilot study to test metformin in young patients with acquired brain damage, either as result of treating a childhood brain tumor or from a traumatic head injury.

We will report back to you with results, as they are published.

Sexy Treatment for Traumatic Brain Injury?

Traumatic Brain Injury (TBI) is a serious public health problem that affects more than 1.7 million Americans each year.  It is the leading cause of death and disability in children and young adults worldwide and is responsible for more than 52,000 deaths, 275,000 hospitalizations, and 80,000 cases of long-term disability in the US alone.

Caused by a bump, blow or jolt to the head or a penetrating head injury, a TBI disrupts the normal function of the brain. The severity of a TBI may range from “mild” – a brief change in mental status or consciousness to “severe” – an extended period of unconsciousness or amnesia after the injury.  Contrary to most media portrayals, the leading cause of TBI is not injuries sustained during war or football, but road traffic accidents.

Despite significant efforts over the past 20 years, there is still no approved treatment to reverse the damage caused by TBI. However, there is a growing body of research to suggest that the sex hormone progesterone may be a powerful neurosteroid that can protect against the effects of TBI.

Discovery of progesterone’s neuroprotective properties began with the observation of a gender difference in response to experimentally induced traumatic brain injury (TBI). After noting anecdotal reports that female rats recover better than male rats following TBI, researchers conducted studies which showed that the hormone might account for this discrepancy in outcomes.

And while progesterone initially may seem like an unusual choice as a neuroprotective therapeutic agent, there is abundant evidence suggesting that this is a normal role of this steroid hormone. In addition to its well-known effects on the reproductive system, progesterone is a potent neurosteroid.  Progesterone receptors are abundant and widely distributed in the central nervous system. Moreover, unlike other sex steroids, progesterone is not only synthesized in the gonads and adrenal glands, but also produced by glial cells in the brain and by Schwann cells in the peripheral nervous system.

BHR Pharma, based in Herndon VA, has developed BHR-100, a novel intravenous formulation of progesterone for treating TBI.   The company’s Phase III multi-center SyNAPSe clinical trial is currently underway and has enrolled 500 of the 1,180 patients suffering from severe TBI needed.

BHR Pharma, is also studying BHR-310 (intranasal progesterone powder) a ready-for-use, nasal spray TBI treatment.  The compound is being developed so that it may be administered to wounded warriors at the site of injury, without the help of medically trained personnel. Studies of progesterone for TBI show that the earlier progesterone is given the better the chance to prevent further damage and improve recovery. A simple nasal spray device would therefore allow the progesterone to begin working before patients are admitted to a hospital.

The FDA has granted Orphan Drug designation to BHR-100 and the drug is on a Fast Track status designed to accelerate its potential approval.  SRxA’s Word on Health will continue to follow these developments and bring you news as it breaks.