A de novo loss-of-function GRIN2A mutation associated with childhood focal epilepsy and acquired epileptic aphasia.
OBJECTIVE
N-methyl-D-aspartate receptors (NMDAR) subunit GRIN2A/GluN2A mutations have been identified in patients with various neurological diseases, such as epilepsy and intellectual disability / developmental delay (ID/DD). In this study, we investigated the phenotype and underlying molecular mechanism of a GRIN2A missense mutation identified by next generation sequencing on idiopathic focal epilepsy using in vitro electrophysiology.


METHODS
Genomic DNA of patients with epilepsy and ID/DD were sequenced by targeted next-generation sequencing within 300 genes related to epilepsy and ID/DD. The effects of one missense GRIN2A mutation on NMDAR function were evaluated by two-electrode voltage clamp current recordings and whole cell voltage clamp current recordings.


RESULTS
We identified one de novo missense GRIN2A mutation (Asp731Asn, GluN2A(D731N)) in a child with unexplained epilepsy and DD. The D731N mutation is located in a portion of the agonist-binding domain (ABD) in the GluN2A subunit, which is the binding pocket for agonist glutamate. This residue in the ABD is conserved among vertebrate species and all other NMDAR subunits, suggesting an important role in receptor function. The proband shows developmental delay as well as EEG-confirmed seizure activity. Functional analyses reveal that the GluN2A(D731N) mutation decreases glutamate potency by over 3,000-fold, reduces amplitude of current response, shortens synaptic-like response time course, and decreases channel open probability, while enhancing sensitivity to negative allosteric modulators, including extracellular proton and zinc inhibition. The combined effects reduce NMDAR function.


CONCLUSIONS
We identified a de novo missense mutation in the GRIN2A gene in a patient with childhood focal epilepsy and acquired epileptic aphasia. The mutant decreases NMDAR activation suggesting NMDAR hypofunction may contribute to the epilepsy pathogenesis.
Department of Pediatrics and Pediatric Epilepsy Center, Peking University First Hospital, Beijing, China.
Kai
K
Gao
Department of Pharmacology, Emory University School of Medicine, Atlanta, GA, United States of America.
Anel
A
Tankovic
Department of Pediatrics and Pediatric Epilepsy Center, Peking University First Hospital, Beijing, China.
Yujia
Y
Zhang
Department of Pharmacology, Emory University School of Medicine, Atlanta, GA, United States of America.
Hirofumi
H
Kusumoto
Department of Pharmacology, Emory University School of Medicine, Atlanta, GA, United States of America.
Jin
J
Zhang
Department of Pharmacology, Emory University School of Medicine, Atlanta, GA, United States of America.|Department of Neurology, Xiangya Hospital, Central South University, Changsha, China.
Wenjuan
W
Chen
Department of Pediatrics and Pediatric Epilepsy Center, Peking University First Hospital, Beijing, China.
Wenshu
W
XiangWei
Department of Pharmacology, Emory University School of Medicine, Atlanta, GA, United States of America.
Gil H
GH
Shaulsky
Department of Pharmacology, Emory University School of Medicine, Atlanta, GA, United States of America.
Chun
C
Hu
Department of Pharmacology, Emory University School of Medicine, Atlanta, GA, United States of America.|Center for Functional Evaluation of Rare Variant (CFERV), Emory University School of Medicine, Atlanta, GA, United States of America.
Stephen F
SF
Traynelis
Department of Pharmacology, Emory University School of Medicine, Atlanta, GA, United States of America.|Center for Functional Evaluation of Rare Variant (CFERV), Emory University School of Medicine, Atlanta, GA, United States of America.
Hongjie
H
Yuan
Department of Pediatrics and Pediatric Epilepsy Center, Peking University First Hospital, Beijing, China.|Center of Epilepsy, Beijing Institute for Brain Disorders, Beijing, China.
Yuwu
Y
Jiang
D019943:Amino Acid Substitution; D004569:Electroencephalography; D004828:Epilepsies, Partial / Q000235:genetics / Q000378:metabolism / Q000503:physiopathology; D005260:Female; D057809:HEK293 Cells; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D018887:Landau-Kleffner Syndrome / Q000235:genetics / Q000378:metabolism / Q000503:physiopathology; D008297:Male; D008564:Membrane Potentials / Q000235:genetics*; D020125:Mutation, Missense; D016194:Receptors, N-Methyl-D-Aspartate / Q000235:genetics / Q000378:metabolism
D016194:Receptors, N-Methyl-D-Aspartate