THC Exposure Of Human iPSC Neurons Impacts Genes Associated With Neuropsychiatric Disorders

Abstract

There is a strong association between cannabis use and schizophrenia but the underlying cellular links are poorly understood. Neurons derived from human-induced pluripotent stem cells (hiPSCs) offer a platform for investigating both baseline and dynamic changes in human neural cells. Here, we exposed neurons derived from hiPSCs to Δ9-tetrahydrocannabinol (THC), and identified diagnosis-specific differences not detectable in vehicle-controls. RNA transcriptomic analyses revealed that THC administration, either by acute or chronic exposure, dampened the neuronal transcriptional response following potassium chloride (KCl)-induced neuronal depolarization. THC-treated neurons displayed significant synaptic, mitochondrial, and glutamate signaling alterations that may underlie their failure to activate appropriately; this blunted response resembles effects previously observed in schizophrenia hiPSC- derived neurons. Furthermore, we show a significant alteration in THC-related genes associated with autism and intellectual disability, suggesting shared molecular pathways perturbed in neuropsychiatric disorders that are exacerbated by THC.

Conflict of interest statement

The authors declare that they have no conflict of interest.

Fig. 1. THC treatment regulates genes involved in mitochondrial and glutamate pathways. 

a RNA sequencing of hiPSC-derived neurons reveals 497 genes (acute) and 810 genes (chronic) are significantly changed following THC exposure, including. b genes involved in mitochondrial (e.g., COX7A2, MT-CO1, and MT-CO3) and glutamate (e.g., GRID2) pathways (Quantitative RT–PCR (qRT–PCR); Ordinary one-way ANOVA with Tukey’s multiple comparisons test: *p < 0.05. n = 5 (see qRT–PCR, Ca–Ce, Supplementary Table S1)). Ingenuity pathway analysis shows that mitochondrial oxidative phosphorylation is strongly altered after both acute c and chronic d THC exposure

Fig. 2. Postsynaptic density and ion channel genes are regulated by THC treatment. 

a, b Multiple postsynaptic density and ion channel genes are significantly altered in hiPSC-derived neurons following acute or chronic THC exposure, including the postsynaptic gene HOMER1 (Quantitative RT–PCR (qRT–PCR); Ordinary one-way ANOVA with Tukey’s multiple comparisons test: *p < 0.05. n = 5 (see qRT–PCR, Ca–Ce, Supplementary Table S1)). c Network analysis combining all THC-related genes from acute and chronic THC treatment shows broad changes to fundamental cellular functions such as RNA biology, chromatin regulation and development

Fig. 3. Genes altered by THC treatment in hiPSC-derived neurons are significantly associated with autism and intellectual disability. 

a Venn diagram showing the overlap between THC-related genes and autism, intellectual disability and schizophrenia. b THC-related genes are significantly related to autism and intellectual disability (p-value < 0.05)

Fig. 4. THC treatment results in neuronal hypo-excitability similar to observations using schizophrenia-associated neurons. 

a Venn diagram showing impaired transcriptional response following 50 mM KCl treatment for 3 h in THC exposure hiPSC-derived neurons. b A similar decrease in significantly regulated transcripts following 50 mM KCl for 3 h is observed in schizophrenia-associated hiPSC-derived neurons. c A cohort of 5 control (C1–5) and 4 schizophrenia-associated (SZ1-4) cases were used for (d) candidate qRT–PCR analysis investigating COX7A2, GRID2 and HOMER1 following acute THC exposure. e Blunted effect of THC treatment can be seen in immediate early gene transcripts such as NR4A1 and (f) FOSB following KCl-induced activation (Quantitative RT–PCR (qRT–PCR); Ordinary one-way ANOVA with Tukey’s multiple comparisons test: *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. n = 5 controls (see qRT–PCR, Ca–Ce, Supplementary Table S1); n = 4 schizophrenia (see qRT–PCR, S1–S4, Supplementary Table S1))
Filed under: Brain and Behaviour,Cannabis/Marijuana,Health :

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