PMID- 28242877
OWN - NLM
STAT- MEDLINE
DCOM- 20180709
LR  - 20230803
IS  - 2045-2322 (Electronic)
IS  - 2045-2322 (Linking)
VI  - 7
IP  - 1
DP  - 2017 Feb 27
TI  - Epilepsy-associated GRIN2A mutations reduce NMDA receptor trafficking and agonist 
      potency - molecular profiling and functional rescue.
PG  - 66
LID - 10.1038/s41598-017-00115-w [doi]
LID - 66
AB  - Mutations in the N-methyl-D-aspartate receptor (NMDAR) gene GRIN2A cause 
      epilepsy-aphasia syndrome (EAS), a spectrum of epileptic, cognitive and language 
      disorders. Using bioinformatic and patient data we shortlisted 10 diverse 
      missense mutations for characterisation. We used high-throughput calcium-flux 
      assays and patch clamp recordings of transiently transfected HEK-293 cells for 
      electrophysiological characterization, and Western blotting and confocal imaging 
      to assay expression and surface trafficking. Mutations P79R, C231Y, G483R and 
      M705V caused a significant reduction in glutamate and glycine agonist potency, 
      whilst D731N was non-responsive. These mutants, along with E714K, also showed 
      significantly decreased total protein levels and trafficking to the cell surface, 
      whilst C436R was not trafficked at all. Crucially this reduced surface expression 
      did not cause the reduced agonist response. We were able to rescue the phenotype 
      of P79R, C231Y, G483R and M705V after treatment with a GluN2A-selective positive 
      allosteric modulator. With our methodology we were not able to identify any 
      functional deficits in mutations I814T, D933N and N976S located between the 
      glutamate-binding domain and C-terminus. We show GRIN2A mutations affect the 
      expression and function of the receptor in different ways. Careful molecular 
      profiling of patients will be essential for future effective personalised 
      treatment options.
FAU - Addis, L
AU  - Addis L
AUID- ORCID: 0000-0001-8455-4278
AD  - Department of Basic and Clinical Neuroscience, Institute of Psychiatry, 
      Psychology and Neuroscience, King's College London, London, UK. 
      laura.addis@kcl.ac.uk.
AD  - Neuroscience Discovery, Eli Lilly Research Centre, Windlesham, Surrey, UK. 
      laura.addis@kcl.ac.uk.
FAU - Virdee, J K
AU  - Virdee JK
AD  - Neuroscience Discovery, Eli Lilly Research Centre, Windlesham, Surrey, UK.
FAU - Vidler, L R
AU  - Vidler LR
AD  - Neuroscience Discovery, Eli Lilly Research Centre, Windlesham, Surrey, UK.
FAU - Collier, D A
AU  - Collier DA
AUID- ORCID: 0000-0003-4087-1559
AD  - Neuroscience Discovery, Eli Lilly Research Centre, Windlesham, Surrey, UK.
FAU - Pal, D K
AU  - Pal DK
AD  - Department of Basic and Clinical Neuroscience, Institute of Psychiatry, 
      Psychology and Neuroscience, King's College London, London, UK.
FAU - Ursu, D
AU  - Ursu D
AD  - Neuroscience Discovery, Eli Lilly Research Centre, Windlesham, Surrey, UK.
LA  - eng
GR  - UC2 HL103010/HL/NHLBI NIH HHS/United States
GR  - RC2 HL102926/HL/NHLBI NIH HHS/United States
GR  - RC2 HL102924/HL/NHLBI NIH HHS/United States
GR  - MR/N026063/1/MRC_/Medical Research Council/United Kingdom
GR  - RC2 HL103010/HL/NHLBI NIH HHS/United States
GR  - RC2 HL102923/HL/NHLBI NIH HHS/United States
GR  - UC2 HL102926/HL/NHLBI NIH HHS/United States
GR  - UC2 HL102923/HL/NHLBI NIH HHS/United States
GR  - UC2 HL102924/HL/NHLBI NIH HHS/United States
GR  - RC2 HL102925/HL/NHLBI NIH HHS/United States
GR  - UC2 HL102925/HL/NHLBI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20170227
PL  - England
TA  - Sci Rep
JT  - Scientific reports
JID - 101563288
RN  - 0 (Glutamates)
RN  - 0 (Receptors, N-Methyl-D-Aspartate)
RN  - SY7Q814VUP (Calcium)
RN  - TE7660XO1C (Glycine)
RN  - VH92ICR8HX (N-methyl D-aspartate receptor subtype 2A)
SB  - IM
MH  - Blotting, Western
MH  - Calcium/metabolism
MH  - Epilepsy/*genetics/physiopathology
MH  - Gene Expression Profiling
MH  - Glutamates/metabolism
MH  - Glycine/metabolism
MH  - HEK293 Cells
MH  - Humans
MH  - Microscopy, Confocal
MH  - *Mutation, Missense
MH  - Patch-Clamp Techniques
MH  - Receptors, N-Methyl-D-Aspartate/*agonists/genetics
PMC - PMC5427847
COIS- The authors declare no competing financial interests.
EDAT- 2017/03/01 06:00
MHDA- 2018/07/10 06:00
PMCR- 2017/02/27
CRDT- 2017/03/01 06:00
PHST- 2016/09/29 00:00 [received]
PHST- 2017/02/06 00:00 [accepted]
PHST- 2017/03/01 06:00 [entrez]
PHST- 2017/03/01 06:00 [pubmed]
PHST- 2018/07/10 06:00 [medline]
PHST- 2017/02/27 00:00 [pmc-release]
AID - 10.1038/s41598-017-00115-w [pii]
AID - 115 [pii]
AID - 10.1038/s41598-017-00115-w [doi]
PST - epublish
SO  - Sci Rep. 2017 Feb 27;7(1):66. doi: 10.1038/s41598-017-00115-w.