Autism spectrum disorder (ASD) is a highly prevalent group of neurodevelopmental disorders whose treatment efficacy is limited by poor understanding of its causal molecular mechanisms. Identifying disease mechanisms in ASD involves overcoming several challenges. First, it is difficult to demonstrate causality for a given mutation, since many mutations increase risk but do not always produce ASD. Second, it is challenging to identify animal models with autism-related phenotypes that are robust across multiple genetic backgrounds. Third, limited understanding of the neuronal subtypes and circuits responsible for behavior is a barrier to studying molecular mechanisms in a disease-relevant cellular context. The proposed research meets all three of these challenges. In the F99 phase, human families with a highly penetrant form of recessive autism caused by mutations in ACTL6B, a neuronal-specific subunit of the BAF chromatin remodeling complex, were identified. Actl6b-/- mice were tested as a model for ACTL6B mutant ASD and found to exhibit autism-related behaviors on two genetic backgrounds and similar brain anatomy to the affected humans. Transcriptional analysis of Actl6b-/- cortical cultures indicated that neural activity-induced genes were de-repressed in the absence of Actl6b, even when action potentials were blocked. The elevated expression of early response genes, including AP1 transcription factors (e.g., Fos, Junb), in Actl6b-/- neurons was associated with increased chromatin accessibility at AP1 sites and activity-related transcriptional changes in late-response genes, implicating abnormal early response gene activation as a potential disease mechanism. The genomic localization of the BAF complex, AP1 transcription factors, and the NCoR complex, which interacted with BAF in cortical tissue, will be studied in wildtype and Actl6b-/- neurons to learn if altered targeting of these complexes may contribute to disease-related transcriptional changes. To gain insight into the affected neuronal circuitry, a serotonin receptor 1b (5HT1b) agonist that was shown to rescue social behavior in the 16p11 autism mouse model (PMID: 30089910) was tested and found to rescue social impairments in Actl6b-/- mice. Serotonergic neuron-specific deletion of Arid1b, the most frequently mutated BAF subunit in autism, caused social impairments in mice that could likewise be rescued with the 5HT1b agonist, indicating that BAF function in serotonergic neurons is critical for social behavior. These studies have inspired the postdoctoral (K00) research direction: to interrogate autism-related molecular mechanisms within neuronal populations that control behavior. The postdoctoral training in systems neuroscience will buoy future studies linking the functions of chromatin regulatory proteins directly to behavior. This research supports the missions of the NIH Blueprint and BRAIN Initiative by providing new tools for autism research and revealing molecular and circuit mechanisms that influence behavior.
Better understanding of causative mechanisms in autism spectrum disorder (ASD) is urgently needed to inform treatment for more than 1% of children worldwide who are affected with ASD. This research identifies mutations in the neuronal BAF chromatin remodeling subunit ACTL6B as a highly penetrant cause of recessive autism and validates Actl6b knockout mice as a model. Building on these findings, the goals of the project are to uncover molecular and circuit mechanisms that cause autism-related phenotypes in Actl6b-/- mice.
