Functional analysis of rare variants found in schizophrenia implicates a critical role for GIT1–PAK3 signaling in neuroplasticity

M. J. Kim, J. Biag, D. M. Fass, M. C. Lewis, Q. Zhang, M. Fleishman, Shanti Gangwar, M. Machius, M. Fromer, S. M. Purcell, S. A. McCarroll, Gabrielle Rudenko, R. T. Premont, E. M. Scolnick, S. J. Haggarty

Research output: Contribution to journalArticle

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Abstract

Although the pathogenesis of schizophrenia (SCZ) is proposed to involve alterations of neural circuits via synaptic dysfunction, the underlying molecular mechanisms remain poorly understood. Recent exome sequencing studies of SCZ have uncovered numerous single-nucleotide variants (SNVs); however, the majority of these SNVs have unknown functional consequences, leaving their disease relevance uncertain. Filling this knowledge gap requires systematic application of quantitative and scalable assays to assess known and novel biological functions of genes. Here we demonstrate loss-of-function effects of multiple rare coding SNVs found in SCZ subjects in the GIT1 (G protein-coupled receptor kinase interacting ArfGAP 1) gene using functional cell-based assays involving coexpression of GIT1 and PAK3 (p21 protein (Cdc42/Rac)-activated kinase 3). Most notably, a GIT1-R283W variant reported in four independent SCZ cases was defective in activating PAK3 as well as MAPK (mitogen-activated protein kinase). Similar functional deficits were found for a de novo SCZ variant GIT1-S601N. Additional assays revealed deficits in the capacity of GIT1-R283W to stimulate PAK phosphorylation in cultured hippocampal neurons. In addition, GIT1-R283W showed deficits in the induction of GAD1 (glutamate decarboxylase 1) protein expression. Extending these functional assays to 10 additional rare GIT1 variants revealed the existence of an allelic series with the majority of the SCZ case variants exhibiting loss of function toward MAPK activation in a manner correlated with loss of PAK3 activation. Taken together, we propose that rare variants in GIT1, along with other genetic and environmental factors, cause dysregulation of PAK3 leading to synaptic deficits in SCZ.Molecular Psychiatry advance online publication, 26 July 2016; doi:10.1038/mp.2016.98.

Original languageEnglish (US)
JournalMolecular Psychiatry
DOIs
StateAccepted/In press - Jul 26 2016

Fingerprint

Neuronal Plasticity
Schizophrenia
Nucleotides
Mitogen-Activated Protein Kinases
cdc42 GTP-Binding Protein
rac GTP-Binding Proteins
G-Protein-Coupled Receptor Kinases
Exome
Genes
Psychiatry
Publications
Phosphotransferases
Phosphorylation
Neurons

ASJC Scopus subject areas

  • Molecular Biology
  • Psychiatry and Mental health
  • Cellular and Molecular Neuroscience

Cite this

Functional analysis of rare variants found in schizophrenia implicates a critical role for GIT1–PAK3 signaling in neuroplasticity. / Kim, M. J.; Biag, J.; Fass, D. M.; Lewis, M. C.; Zhang, Q.; Fleishman, M.; Gangwar, Shanti; Machius, M.; Fromer, M.; Purcell, S. M.; McCarroll, S. A.; Rudenko, Gabrielle; Premont, R. T.; Scolnick, E. M.; Haggarty, S. J.

In: Molecular Psychiatry, 26.07.2016.

Research output: Contribution to journalArticle

Kim, MJ, Biag, J, Fass, DM, Lewis, MC, Zhang, Q, Fleishman, M, Gangwar, S, Machius, M, Fromer, M, Purcell, SM, McCarroll, SA, Rudenko, G, Premont, RT, Scolnick, EM & Haggarty, SJ 2016, 'Functional analysis of rare variants found in schizophrenia implicates a critical role for GIT1–PAK3 signaling in neuroplasticity', Molecular Psychiatry. https://doi.org/10.1038/mp.2016.98
Kim, M. J. ; Biag, J. ; Fass, D. M. ; Lewis, M. C. ; Zhang, Q. ; Fleishman, M. ; Gangwar, Shanti ; Machius, M. ; Fromer, M. ; Purcell, S. M. ; McCarroll, S. A. ; Rudenko, Gabrielle ; Premont, R. T. ; Scolnick, E. M. ; Haggarty, S. J. / Functional analysis of rare variants found in schizophrenia implicates a critical role for GIT1–PAK3 signaling in neuroplasticity. In: Molecular Psychiatry. 2016.
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abstract = "Although the pathogenesis of schizophrenia (SCZ) is proposed to involve alterations of neural circuits via synaptic dysfunction, the underlying molecular mechanisms remain poorly understood. Recent exome sequencing studies of SCZ have uncovered numerous single-nucleotide variants (SNVs); however, the majority of these SNVs have unknown functional consequences, leaving their disease relevance uncertain. Filling this knowledge gap requires systematic application of quantitative and scalable assays to assess known and novel biological functions of genes. Here we demonstrate loss-of-function effects of multiple rare coding SNVs found in SCZ subjects in the GIT1 (G protein-coupled receptor kinase interacting ArfGAP 1) gene using functional cell-based assays involving coexpression of GIT1 and PAK3 (p21 protein (Cdc42/Rac)-activated kinase 3). Most notably, a GIT1-R283W variant reported in four independent SCZ cases was defective in activating PAK3 as well as MAPK (mitogen-activated protein kinase). Similar functional deficits were found for a de novo SCZ variant GIT1-S601N. Additional assays revealed deficits in the capacity of GIT1-R283W to stimulate PAK phosphorylation in cultured hippocampal neurons. In addition, GIT1-R283W showed deficits in the induction of GAD1 (glutamate decarboxylase 1) protein expression. Extending these functional assays to 10 additional rare GIT1 variants revealed the existence of an allelic series with the majority of the SCZ case variants exhibiting loss of function toward MAPK activation in a manner correlated with loss of PAK3 activation. Taken together, we propose that rare variants in GIT1, along with other genetic and environmental factors, cause dysregulation of PAK3 leading to synaptic deficits in SCZ.Molecular Psychiatry advance online publication, 26 July 2016; doi:10.1038/mp.2016.98.",
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