April 11, 2011
Gene linked to severity of autism’s social dysfunction
With the help of two sets of brothers with autism, Johns Hopkins scientists have identified a gene associated with autism that appears to be linked specifically to the severity of social-interaction deficits.
The gene, known as GRIP1 for glutamate receptor interacting protein 1, is a blueprint for a traffic-directing protein at synapses—those specialized contact points between brain cells across which chemical signals flow.
Identified more than a decade ago by Richard L. Huganir, professor and director of the Solomon H. Snyder Department of Neuroscience at the Johns Hopkins University School of Medicine, and a Howard Hughes Medical Institute investigator, GRIP1 regulates how fast receptors travel to a cell’s surface, where they are activated by a brain-signaling chemical called glutamate, allowing neurons to communicate with one another.
The new study, which tracked two versions of GRIP1 in the genomes of 480 people with autism, was published March 22 in the Proceedings of the National Academy of Sciences and lends support to a prevailing theory that autism spectrum disorders, molecularly speaking, reflect an imbalance between inhibitory and excitatory signaling at synapses.
“The GRIP1 variants we studied are not sufficient to cause autism by themselves but appear to be contributing factors that can modify the severity of the disease,” said Tao Wang, an assistant professor in the McKusick-Nathans Institute of Genetic Medicine at the Johns Hopkins School of Medicine. “GRIP1 mutations seem to contribute to social interaction deficits in the patients we studied.”
The researchers examined a part of the genomes of 480 patients with autism and compared them with those of 480 people of similar ethnicity without the disorder. Analyzing about 50 genes known to make proteins involved in a brain-signaling pathway, they ultimately focused their investigation on GRIP1, a protein found at both inhibitory and excitatory synapses, according to Wang.
Initially, looking under a microscope at normal mouse neurons and at neurons with a mutant version of GRIP1, the investigators marked the receptor proteins with green fluorescence, added a chemical that promotes their “disappearance” deep inside a cell and timed the rates at which they disappeared—leaving a cell unable to respond to signals from other cells. They also timed the re-emergence of the protein on the cell surface. With the GRIP1 mutant neurons, the receptors recycled to the surface twice as fast as in the normal neurons.
“If the receptors are recycling faster, the number of receptors on the surface is greater, so the cells are more sensitive to glutamate,” Huganir said. “The quicker the recycling, the more receptors on the surface and the stronger the excitatory transmission.”
Even if just the excitatory synapses are affected and the inhibitory ones don’t change, that alone affects the relative balance of signaling, Huganir said.
Next, using 10 mice genetically engineered to lack both normal and mutant GRIP proteins, researchers watched what happened when each animal was put into a box where it could choose between spending time with a mouse it hadn’t encountered before or with an inanimate object. They compared the behaviors of these mice with those of 10 normal mice put into the same social situation. Mice lacking both GRIP1 and GRIP2 spent twice as much time as wild-type (normal) mice interacting with other mice as they did with inanimate objects.
“These results support a role for GRIP1 in social behavior and implicate its variants in modulating autistic behavior,” Wang said.
Finally, the team looked at the behavioral analyses of individuals in two families, each with two autistic brothers, and correlated their scores on standard diagnostic tests that assessed social interaction with their genotypes for GRIP1 variants.
In one family, the brother with two copies of the GRIP1 mutant variety scored lower on social-interaction tests than his brother who had only one copy of the GRIP1 variant. The boys’ mother, although not diagnosed as autistic, had a history of restricted interests, poor eye contact and repetitive behavior. Tests showed that she also carried one copy of the variant.
In a second family, the autistic brother with one copy of the GRIP1 variant had lower social-interaction scores than his autistic sibling without a GRIP1 variant.
Because the GRIP1 gene resides in synapses where other genes also implicated in autism have been found, this location is potentially important in terms of clinical relevance, Huganir said. The team plans to sequence hundreds more synaptic proteins in autistic patients to look for mutations and then follow up with functional analyses.
This study was supported in part by research grants from the Autism Speaks Foundation and National Institute of Child Health and Human Development.
Authors on the paper from Johns Hopkins, in addition to Huganir and Wang, are Rebeca Mejias, Abby Adamczyk, Victor Anggono, Tejasvi Niranjan, Gareth M. Thomas, Kamal Sharma, M. Daniele Fallin, Walter E. Kaufmann, Mikhail Pletnikov and David Valle.
Cindy Skinner, Charles E. Schwartz and Roger Stevenson, all of the Greenwood Genetic Center, are also authors on the paper.