Study suggests new molecular strategy for treating fragile X syndrome

Building on more than two decades of research, a study by MIT neuroscientists at The Picower Institute for Learning and Memory reports a new way to treat pathology and symptoms of fragile X syndrome, the most common genetically-caused autism spectrum disorder. The team showed that augmenting a novel type of neurotransmitter signaling reduced hallmarks of fragile X in mouse models of the disorder.

The new approach described in Cell Reports works by targeting a specific molecular subunit of  “NMDA” receptors that they discovered plays a key role in how neurons synthesize proteins to regulate their connections, or “synapses,” with other neurons in brain circuits. The scientists showed that in fragile X model mice, increasing the receptor’s activity caused neurons in the hippocampus region of the brain to increase molecular signaling that suppressed excessive bulk protein synthesis, leading to other key improvements.

Setting the table

“One of the things I find most satisfying about this study is that the pieces of the puzzle fit so nicely into what had come before,” said study senior author Mark Bear, Picower Professor in MIT’s Department of Brain and Cognitive Sciences. Former postdoc Stephanie Barnes, now a lecturer at the University of Glasgow, is the study’s lead author.

Bear’s lab studies how neurons continually edit their circuit connections, a process called “synaptic plasticity” that scientists believe to underlie the brain’s ability to adapt to experience and to form and process memories. These studies led to two discoveries that set the table for the newly published advance. In 2011, Bear’s lab showed that fragile X and another autism disorder, tuberous sclerosis (Tsc), represented two ends of a continuum of a kind of protein synthesis in the same neurons. In fragile X there was too much. In Tsc there was too little. When lab members crossbred fragile X and Tsc mice, in fact, their offspring emerged healthy as the mutations of each disorder essentially canceled each other out. 

More recently, Bear’s lab showed a different dichotomy. It has long been understood from their seminal work in the 1990s that the flow of calcium ions through NMDA receptor can trigger a form of synaptic plasticity called “long-term depression” (LTD). But in 2020, they found that another mode of signaling by the receptor—one that did not require ion flow—altered protein synthesis in the neuron and caused a physical shrinking of the dendritic “spine” structures housing synapses.

For Bear and Barnes these studies raised the prospect that if they could pinpoint how NMDA receptors affect protein synthesis they might identify a new mechanism that could be manipulated therapeutically to address fragile X (and perhaps tuberous sclerosis) pathology and symptoms. That would be an important advance to complement ongoing work Bear’s lab has done to correct fragile X protein synthesis levels via another receptor called mGluR5.

Receptor dissection

In the new study, Bear and Barnes’s team decided to use the non-ionic effect on spine shrinkage as a readout to dissect how NMDARs signal protein synthesis for synaptic plasticity in hippocampus neurons. They hypothesized that the dichotomy of ionic effects on synaptic function and non-ionic effects on spine structure might derive from the presence of two distinct components of NMDAR receptors: “subunits” called GluN2A and GluN2B. To test that, they used genetic manipulations to knock out each of the subunits. When they did so, they found that knocking out “2A” or “2B” could eliminate LTD but that only knocking out 2B affected spine size. Further experiments clarified that 2A and 2B are required for LTD, but that spine shrinkage solely depends on the 2B subunit.

The next task was to resolve how the 2B subunit signals spine shrinkage. A promising possibility was a part of the subunit called the “carboxyterminal domain,” or CTD. So, in a new experiment Bear and Barnes took advantage of a mouse that had been genetically engineered by researchers at the University of Edinburgh so that the 2A and 2B CTDs could be swapped with one another. A telling result was that when the 2B subunit lacked its proper CTD, the effect on spine structure disappeared. The result affirmed that the 2B subunit signals spine shrinkage via its CTD.

Another consequence of replacing the CTD of the 2B subunit was an increase in bulk protein synthesis that resembled findings in fragile X.  Conversely, augmenting the non-ionic signaling through the 2B subunit suppressed bulk protein synthesis, reminiscent of Tsc.

Treating fragile X

Putting the pieces together, the findings indicated that augmenting signaling through the 2B subunit might, like introducing the mutation causing Tsc, rescue aspects of fragile X.

Indeed when the scientists swapped in the 2B subunit CTD of NMDA receptor in fragile X model mice they found correction of not only the excessive bulk protein synthesis, but also altered synaptic plasticity, and increased electrical excitability that are hallmarks of the disease. To see if a treatment that targets NMDA receptors might be effective in fragile X, they tried an experimental drug called Glyx-13. This drug binds to the 2B subunit of NMDA receptors to augment signaling. The researchers found that this treatment can also normalize protein synthesis and reduced sound-induced seizures in the fragile X mice. 

The team now hypothesizes, based on another prior study in the lab, that the beneficial effect to fragile X mice of the 2B subunit’s CTD signaling is that it shifts the balance of protein synthesis away from an all-too-efficient translation of short messenger RNAs (which leads to excessive bulk protein synthesis) toward a lower efficiency translation of longer messenger RNAs.

Bear said he does not know what the prospects are for Glyx-13 as a clinical drug, but he noted that there are some drugs in clinical development that specifically target the 2B subunit of NMDA receptors.

In addition to Bear and Barnes, the study’s other authors are Aurore Thomazeau, Peter Finnie, Max Heinreich, Arnold Heynen, Noboru Komiyama, Seth Grant, Frank Menniti and Emily Osterweil.

The FRAXA Foundation, The Picower Institute for Learning and Memory, The Freedom Together Foundation and the National Institutes of Health funded the study.