TY - JOUR
T1 - GLS hyperactivity causes glutamate excess, infantile cataract and profound developmental delay
AU - Rumping, Lynne
AU - Tessadori, Federico
AU - Pouwels, Petra J W
AU - Vringer, Esmee
AU - Wijnen, Jannie P
AU - Bhogal, Alex A
AU - Savelberg, Sanne M C
AU - Duran, Karen J
AU - Bakkers, Mark J G
AU - Ramos, Rúben J J
AU - Schellekens, Peter A W
AU - Kroes, Hester Y
AU - Klomp, Dennis W J
AU - Black, Graeme C M
AU - Taylor, Rachel L
AU - Bakkers, Jeroen P W
AU - Prinsen, Hubertus C M T
AU - van der Knaap, Marjo S
AU - Dansen, Tobias B
AU - Rehmann, Holger
AU - Zwartkruis, Fried J T
AU - Houwen, Roderick H J
AU - van Haaften, Gijs
AU - Verhoeven-Duif, Nanda M
AU - Jans, Judith J M
AU - van Hasselt, Peter M
PY - 2019/1/1
Y1 - 2019/1/1
N2 - Loss-of-function mutations in glutaminase (GLS), the enzyme converting glutamine into glutamate, and the counteracting enzyme glutamine synthetase (GS) cause disturbed glutamate homeostasis and severe neonatal encephalopathy. We report a de novo Ser482Cys gain-of-function variant in GLS encoding GLS associated with profound developmental delay and infantile cataract. Functional analysis demonstrated that this variant causes hyperactivity and compensatory downregulation of GLS expression combined with upregulation of the counteracting enzyme GS, supporting pathogenicity. Ser482Cys-GLS likely improves the electrostatic environment of the GLS catalytic site, thereby intrinsically inducing hyperactivity. Alignment of +/-12.000 GLS protein sequences from >1000 genera revealed extreme conservation of Ser482 to the same degree as catalytic residues. Together with the hyperactivity, this indicates that Ser482 is evolutionarily preserved to achieve optimal-but submaximal-GLS activity. In line with GLS hyperactivity, increased glutamate and decreased glutamine concentrations were measured in urine and fibroblasts. In the brain (both grey and white matter), glutamate was also extremely high and glutamine was almost undetectable, demonstrated with magnetic resonance spectroscopic imaging at clinical field strength and subsequently supported at ultra-high field strength. Considering the neurotoxicity of glutamate when present in excess, the strikingly high glutamate concentrations measured in the brain provide an explanation for the developmental delay. Cataract, a known consequence of oxidative stress, was evoked in zebrafish expressing the hypermorphic Ser482Cys-GLS and could be alleviated by inhibition of GLS. The capacity to detoxify reactive oxygen species was reduced upon Ser482Cys-GLS expression, providing an explanation for cataract formation. In conclusion, we describe an inborn error of glutamate metabolism caused by a GLS hyperactivity variant, illustrating the importance of balanced GLS activity.
AB - Loss-of-function mutations in glutaminase (GLS), the enzyme converting glutamine into glutamate, and the counteracting enzyme glutamine synthetase (GS) cause disturbed glutamate homeostasis and severe neonatal encephalopathy. We report a de novo Ser482Cys gain-of-function variant in GLS encoding GLS associated with profound developmental delay and infantile cataract. Functional analysis demonstrated that this variant causes hyperactivity and compensatory downregulation of GLS expression combined with upregulation of the counteracting enzyme GS, supporting pathogenicity. Ser482Cys-GLS likely improves the electrostatic environment of the GLS catalytic site, thereby intrinsically inducing hyperactivity. Alignment of +/-12.000 GLS protein sequences from >1000 genera revealed extreme conservation of Ser482 to the same degree as catalytic residues. Together with the hyperactivity, this indicates that Ser482 is evolutionarily preserved to achieve optimal-but submaximal-GLS activity. In line with GLS hyperactivity, increased glutamate and decreased glutamine concentrations were measured in urine and fibroblasts. In the brain (both grey and white matter), glutamate was also extremely high and glutamine was almost undetectable, demonstrated with magnetic resonance spectroscopic imaging at clinical field strength and subsequently supported at ultra-high field strength. Considering the neurotoxicity of glutamate when present in excess, the strikingly high glutamate concentrations measured in the brain provide an explanation for the developmental delay. Cataract, a known consequence of oxidative stress, was evoked in zebrafish expressing the hypermorphic Ser482Cys-GLS and could be alleviated by inhibition of GLS. The capacity to detoxify reactive oxygen species was reduced upon Ser482Cys-GLS expression, providing an explanation for cataract formation. In conclusion, we describe an inborn error of glutamate metabolism caused by a GLS hyperactivity variant, illustrating the importance of balanced GLS activity.
KW - Adolescent
KW - Animals
KW - Brain/metabolism
KW - Cataract/genetics
KW - Child, Preschool
KW - Developmental Disabilities/genetics
KW - Disease Models, Animal
KW - Female
KW - Fibroblasts
KW - Gain of Function Mutation/genetics
KW - Glutamate-Ammonia Ligase/genetics
KW - Glutamic Acid/genetics
KW - Glutaminase/genetics
KW - Glutamine/metabolism
KW - HEK293 Cells
KW - Humans
KW - Male
KW - Oxidative Stress
KW - Reactive Oxygen Species/metabolism
KW - Zebrafish
UR - https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=85058773908&origin=inward
UR - https://www.ncbi.nlm.nih.gov/pubmed/30239721
U2 - https://doi.org/10.1093/hmg/ddy330
DO - https://doi.org/10.1093/hmg/ddy330
M3 - Article
C2 - 30239721
SN - 0964-6906
VL - 28
SP - 96
EP - 104
JO - Human Molecular Genetics
JF - Human Molecular Genetics
IS - 1
ER -