Neuron-specific transcriptomic signatures indicate neuroinflammation and altered neuronal activity in ASD temporal cortex

Autism spectrum disorder (ASD) is a highly heterogeneous disorder, yet transcriptomic profiling of bulk brain tissue has identified substantial convergence among dysregu-lated genes and pathways in ASD. However, this approach lacks cell-specific resolution. We performed comprehensive transcriptomic analyses on bulk tissue and laser-capture microdissected (LCM) neurons from 59 postmortem human brains (27 ASD and 32 controls) in the superior temporal gyrus (STG) of individuals ranging from 2 to 73 years of age. In bulk tissue, synaptic signaling, heat shock protein-related pathways, and RNA splicing were significantly altered in ASD. There was age-dependent dysregulation of genes involved in gamma aminobutyric acid (GABA) (GAD1 and GAD2) and glutamate (SLC38A1) signaling pathways. In LCM neurons, AP-1-mediated neuroinflammation and insulin/IGF-1 signaling pathways were upregulated in ASD, while mitochondrial function, ribosome, and spliceosome components were downregulated. GABA synthesiz-ing enzymes GAD1 and GAD2 were both downregulated in ASD neurons. Mechanistic modeling suggested a direct link between inflammation and ASD in neurons, and pri-oritized inflammation-associated genes for future study. Alterations in small nucleolar RNAs (snoRNAs) associated with splicing events suggested interplay between snoRNA dysregulation and splicing disruption in neurons of individuals with ASD. Our findings supported the fundamental hypothesis of altered neuronal communication in ASD, demonstrated that inflammation was elevated at least in part in ASD neurons, and may reveal windows of opportunity for biotherapeutics to target the trajectory of gene expression and clinical manifestation of ASD throughout the human lifespan.SignificanceWe present a comprehensive assessment of neuronal cell-type -specific gene expression and alternative splicing changes in ASD cortex, directly comparing RNA-seq results from bulk tissue with isolated neurons. We observe strong signatures of cell stress and neural-immune/ inflammatory pathway activation present within ASD neurons-a signal that is typically attributed to astrocyte/microglial populations. Our findings also provide further evidence for the hypothesized imbalance of excitatory to inhibitory neuronal activity in the brains of individuals with ASD. Moreover, we find that the transcriptomic architecture of ASD interacts substantially with age, thus revealing windows of opportunity for treatments that target specific molecular pathology.

Automated summary

Autism spectrum disorder (ASD) is a heterogeneous disorder with gene and pathway dysregulation in the brain. Transcriptomic analyses were performed on both bulk tissue and laser-capture microdissected neurons from postmortem human brains. Dysregulation of synaptic signaling, inflammation, and RNA splicing were observed in ASD. Alterations in GABA and glutamate signaling pathways were age-dependent. Moreover, dysregulation of inflammatory pathways and splicing disruption were found in ASD neurons.