Analysis of genes within the schizophrenia-linked 22q11.2 deletion identifies interaction of night owl/LZTR1 and NF1 in GABAergic sleep control

The human 22q11.2 chromosomal deletion is one of the strongest identified genetic risk factors for schizophrenia. Although the deletion spans a number of known genes, the contribution of each of these to the 22q11.2 deletion syndrome (DS) is not known. To investigate the effect of individual genes within this interval on the pathophysiology associated with the deletion, we analyzed their role in sleep, a behavior affected in virtually all psychiatric disorders, including the 22q11.2 DS. We identified the gene LZTR1 (night owl, nowl) as a regulator of night-time sleep in Drosophila. In humans, LZTR1 has been associated with Ras-dependent neurological diseases also caused by Neurofibromin-1 (Nf1) deficiency. We show that Nf1 loss leads to a night-time sleep phenotype nearly identical to that of nowl loss and that nowl negatively regulates Ras and interacts with Nf1 in sleep regulation. Furthermore, nowl is required for metabolic homeostasis, suggesting that LZTR1 may contribute to the genetic susceptibility to obesity associated with the 22q11.2 DS. Knockdown of nowl or Nf1 in GABA-responsive sleep-promoting neurons elicits the sleep phenotype, and this defect can be rescued by increased GABA(A) receptor signaling, indicating that Nowl regulates sleep through modulation of GABA signaling. Our results suggest that nowl/LZTR1 may be a conserved regulator of GABA signaling important for normal sleep that contributes to the 22q11.2 DS. Author summary Schizophrenia is a devastating mental disorder with a large genetic component to disease predisposition. One of the strongest genetic risk factors for this disorder is a relatively small genetic deletion of 43 genes on the 22(nd) chromosome, called 22q11.2, which confers about a 25% risk of schizophrenia development. However, it is likely that only some of these deleted genes affect disease risk, so we tested most of them individually. One of the main symptoms of schizophrenia is disturbed sleep. Sleep is an evolutionarily conserved behavior that can be easily studied in the fruit fly Drosophila melanogaster, so we investigated the effect on sleep of blocking expression of the fly homologs of most of the 22q11.2 genes and identified the gene LZTR1 (night owl, nowl) as an important sleep regulator. We found that Nowl/LZTR1 is required for inhibition of the Ras pathway and interacts genetically with the Ras inhibitor NF1. Nowl/LZTR1 appears to function in sleep by modulating inhibitory GABA signaling, which is affected in schizophrenia. Thus, this gene may underlie some of the phenotypes of the human schizophrenia-risk deletion.