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

Molecular Psychiatry volume 22, pages 417–429 (2017)Cite this article

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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.

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Acknowledgements

We thank members of the Stanley Center for Psychiatric Research and the Chemical Neurobiology Laboratory for helpful discussions and critical feedback. Dr Guoping Feng (MIT) is thanked for critical feedback. This work was supported by funding from the Stanley Medical Research Institute and the National Institute of Mental Health (R01MH095088 to SJH).

Author contributions

MJK JB, ML, QZ, EMS and SJH conceived and designed the experiments. MJK, JB, MCL, DMF and SJH performed the experiments and/or analyzed data. RTP assisted with data interpretation and edited the manuscript. MF, SMP, SAM, EMS and SJH assisted with analysis of human genetic data. SPG, MM and GR performed homology model analysis. MJK, JB, MCL, DMF, GR, EMS and SJH wrote and edited the manuscript. All authors discussed the results and implications and commented on the final version of the manuscript.

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Author notes

  1. M J Kim

    Present address: 11Current address: Department of Neurology, Northwestern University Feinberg School of Medicine, Chicago, IL, USA.,

  2. J Biag

    Present address: 12Current address: Novartis Pharmaceuticals, Cambridge, MA, USA.,

  3. M J Kim, J Biag and D M Fass: These three authors contributed equally to this work.

Authors and Affiliations

  1. Stanley Center for Psychiatric Research, Broad Institute of MIT and Harvard, Cambridge, MA, USA

    M J Kim, J Biag, D M Fass, M C Lewis, M Fleishman, M Fromer, S M Purcell, S A McCarroll, E M Scolnick & S J Haggarty

  2. Departments of Neurology and Psychiatry, Chemical Neurobiology Laboratory, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA

    D M Fass & S J Haggarty

  3. Center for Human Genetic Research, Massachusetts General Hospital, Boston, MA, USA

    D M Fass, S M Purcell & S J Haggarty

  4. Psychiatric and Neurodevelopmental Genetics Unit, Massachusetts General Hospital, Boston, MA, USA

    D M Fass, S M Purcell & S J Haggarty

  5. Molecular Neurogenetics Unit, Massachusetts General Hospital, Boston, MA, USA

    D M Fass & S J Haggarty

  6. Department of Brain and Cognitive Sciences, McGovern Institute for Brain Research, Massachusetts Institute of Technology, Cambridge, MA, USA

    Q Zhang & M Fleishman

  7. Department of Pharmacology and Toxicology, Sealy Center for Structural Biology and Biophysics, University of Texas Medical Branch, Galveston, TX, USA

    S P Gangwar, M Machius & G Rudenko

  8. Division of Psychiatric Genomics, Department of Psychiatry, Icahn School of Medicine at Mount Sinai, New York, NY, USA

    M Fromer

  9. Department of Medicine, Duke University Medical Center, Durham, NC, USA

    R T Premont

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  1. M J Kim
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Kim, M., Biag, J., Fass, D. et al. Functional analysis of rare variants found in schizophrenia implicates a critical role for GIT1–PAK3 signaling in neuroplasticity. Mol Psychiatry 22, 417–429 (2017). https://doi.org/10.1038/mp.2016.98

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