Dysregulated glutamatergic neurotransmission has been strongly implicated in the psychopathology of schizophrenia. Recent studies have highlighted the therapeutic promise of presynaptic reductions in glutamate transmission. We have shown that neurons from glutaminase (GLS1) knockout mice show an activity-dependent presynaptic reduction in glutamatergic synaptic transmission. While GLS1 knockout mice die shortly after birth, GLS1 haploinsufficient mice with one functional GLS1 allele (GLS1 hets) are remarkably normal. Strikingly, functional imaging reveals that the mice have focal hypometabolism in the hippocampus, mainly involving the CA1 subregion and the subiculum that is the exact inverse of recent imaging findings in patients with schizophrenia. Moreover, when challenged with pro-psychotic drugs, GLS1 het mice manifest behavioral and neurochemical phenotypes consistent with schizophrenia resilience. Thus, reducing glutaminase activity appears to have therapeutic potential for schizophrenia. To translate this discovery to clinical application, we propose testing the hypothesis that GLS1 het mice are in fact resilient to a range of pro-schizophrenic insults. Using tissue-specific GLS1 deletions, we will ask whether the hippocampal hypometabolism arises from the reduction in GLS1 in the hippocampus, and whether this modulation is sufficient to produce the resilience phenotype. To begin to understand the implications of the resilience phenotype for the pathophysiology of schizophrenia, we will identify the synaptic alterations in the hippocampus that underlie the hypoactivity profile. To test the therapeutic potential of GLS1 inhibition directly, we will induce GLS1 haploinsufficiency in adult mice, doing what we term genetic-pharmacotherapy, to investigate the acute and chronic effects of the intervention. We will induce GLS1 haploinsufficiency earlier in development to explore potential benefits of early intervention and neurodevelopmental contributions. Finally, we will do high-throughput screening to identify small-molecule GLS1 inhibitors with nanomolar efficacy as drug candidates. GLS1 inhibition has therapeutic potential not only for schizophrenia, but also for stroke, and other neurodegenerative disorders involving excitotoxicity, so a CNS-active GLS1 inhibitor will likely have broad therapeutic promise. In summary, the planned preclinical studies together with identification of drug candidates should provide the basis for movement of GLS1 inhibition towards clinical trials as a novel pharmacotherapy for schizophrenia. PUBLIC HEALTH RELEVANCE: We have recently generated a coherent set of clinical and basic findings raising the hypothesis that pharmacological inhibition of phosphate-activated glutaminase, the product of gene GLS1, should prove therapeutic in schizophrenia. We propose to test this hypothesis using GLS1-deficient mice as a proof of concept. The aims are to gain insight into the therapeutic basis for GLS1 inhibition, to elucidate the circuitry involved, and to gain the necessary knowledge for moving toward clinical drug development.