PROJECT SUMMARY Poor sleep is common in neurodevelopment disorders such as autism spectrum disorder (ASD), with up to 93% of ASD individuals reporting sleep problems. These problems worsen quality of life and core symptoms of ASD and likely precede an ASD diagnosis, suggesting they start early in life. The potential adverse impact of early life sleep disruption is supported by animal model studies showing long term functional consequences on behavior. However, little is known about the underlying molecular consequences of sleep deprivation (SD) early in life. Our recent work in wildtype mice demonstrates that the cortical transcriptional response to SD is more prominent at postnatal day 24 (P24) with a preferential downregulation on the Wnt-singling pathway compared to adulthood. Nevertheless, the transcriptional response and molecular mechanisms underlying the adverse effects of SD early in life are poorly understood in the context of ASD. My overall goal is to understand the detrimental consequences of sleep loss in neurodevelopmental disorders. Previous work in the lab showed that mice carrying a mutation in the ASD gene Shank3 (Shank3∆C mice) are the only ASD mouse model that replicates the clinical features of insomnia in ASD and show an abnormal transcriptional response to SD in adults. In my graduate work, I used polysomnography in young Shank3∆C mice to show that sleep problems are present and detectable at P24. In the F99 phase, I will investigate how this ASD mutation alters the impact of SD on transcription at P24 using both bulk and single nuclear RNA sequencing. In addition, I will use targeted quantitative proteomic for SHANK3 and -catenin to investigate the effects of Shank3∆C and SD on canonical Wnt-signaling. Nuclear -catenin levels are a well-established marker of canonical Wnt-signaling activation and SHANK3 has been shown to bind -catenin in the nucleus and inhibit its transcriptional activity. My sponsor, Dr. Peixoto has an established track record analyzing transcriptomic response to SD in mouse models and has establishes the Shank3∆C model for the study of sleep in ASD. My co-sponsor, Dr. Frank has extensive experience in developmental neuroscience and has pioneered many of the studies characterizing the role of sleep-in developmental synaptic plasticity. Additional members of my advisory committee, Dr Hicks and Dr. Prasad are world experts on single-cell transcriptomics and proteomics. In the pre-doctoral F99 phase, I will learn to use proteomic and transcriptomics approaches and advanced transcriptomic data analysis. In the postdoctoral K00 phase, I will build upon these skills and use genetic mouse models to integrate circuit level manipulation and multi-omics to better understand the consequences of sleep loss on behavior in neurodevelopmental disorders. Overall, the proposed training will optimally position me to start an independent research career at a leading neuroscience research institute and advance our unders...