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 are just a few of the technologies awaiting professors and students in the laboratory.
Yet the most complex and potentially influential findings to emerge from the school’s College of Science might come down to this: a kid spitting in a cup after football practice.
It happens once a week to the 12- and 13-year-old kids playing for the Jets, an A-League football team in the Central Loudoun Youth Football League. Athletic trainers collect the players’ saliva samples and 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,” from which he eventually extracts the samples and runs them through sophisticated machinery to determine changes in protein variance that no other technique has been able to identify. Comparing each kid’s spit samples to previous submissions, he hopes to uncover a handful of proteins that can detect concussions.
Working alongside Caswell is another Mason professor, 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. 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 - a common practice conducted by scientists all over the world - 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.”
Indeed, 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 percent 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 swells 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 CLYFL remains their only youth league partner, though they hope that more leagues are added in the future when more resources become available (the two have applied for a grant from the GE Head Health Challenge).
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.”
The Student Athlete Protection Act, passed in Virginia in 2011, determined that 1,760 concussions occurred across all sports last year in the Fairfax County Public School system’s 25 high schools. The problem with that legislation, Caswell says, is that it stops with public schools. Caswell and Petricoin’s work with the CLYFL this fall 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.
Even 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. In an area known for making headway on athletic training and health care, he senses his team is well-positioned to make meaningful discoveries that could impact concussion policies on a greater scale.
“The school systems here are very well resourced,” Caswell said. “I think because the standard of care is so high here compared to other areas of the country that we can test some things in this area that eventually will be standard care across the country.”