ABSTRACT Methamphetamine addiction remains a major public health concern in the United States that has paralleled the opioid epidemic. Psychostimulant use disorders have a heritable genetic component that remains unexplained. Methamphetamine targets membrane and vesicular transporters to increase synaptic dopamine, norepinephrine, and serotonin. We previously identified Hnrnph1 (heterogeneous nuclear ribonucleoprotein H1) as a quantitative trait gene underlying methamphetamine behavioral sensitivity. Hnrnph1 encodes the RNA-binding protein hnRNP H1 that is ubiquitously expressed in neurons throughout the brain. Gene-edited mice with a heterozygous frameshift deletion in the first coding exon of Hnrnph1 showed reduced methamphetamine-induced dopamine release and behaviors. To inform the mechanism linking hnRNP H dysfunction with reduced methamphetamine neurobehavioral effects, we surveyed the mRNA targetome of hnRNP H via cross-linking immunoprecipitation coupled with RNA-sequencing in striatal tissue at baseline and at 30 min post-methamphetamine. Methamphetamine induced opposite changes in RNA-binding targets of hnRNP H in Hnrnph1 mutants versus wild-types, including 3’UTR targets in mRNAs enriched for synaptic proteins involved in dopamine release and excitatory synaptic plasticity. Targetome, transcriptome, and spliceome analyses triangulated on a methamphetamine-induced upregulation of the calcium channel subunit transcript Cacna2d2 and decreased its 3’UTR usage in hyposensitive Hnrnph1 mutants. Pretreatment with pregabalin, an inhibitor of α2δ2 and α2δ1 voltage-gated calcium channel subunits attenuated methamphetamine-induced locomotor activity in wild-type females but not in Hnrnph1 mutants, supporting Cacna2d2 as a hnRNP H target. Our study identifies a dynamic hnRNP H RNA targetome that can rapidly and adaptively respond to methamphetamine to regulate gene expression and likely synaptic plasticity and behavior. SIGNIFICANCE STATEMENT The genetic risks mediating psychostimulant addiction are unknown and there are no FDA-approved treatments. We identified Hnrnph1 in modulating methamphetamine behavioral sensitivity in mice. Hnrnph1 codes for hnRNP H1, an RNA-binding protein. Here, we asked whether an Hnrnph1 mutation and methamphetamine treatment would change the hnRNP H RNA targets and whether these targets could tell us how Hnrnph1 is linked to behavior. We identified a calcium channel subunit that is a primary target of the FDA-approved drug pregabalin (a.k.a. Lyrica®). Like the Hnrnph1 mutation, pregabalin reduced methamphetamine behaviors in wild-type mice. We propose hnRNP H regulates calcium channels in response to methamphetamine-induced perturbations in neurotransmitter release. Accordingly, pregabalin could represent a novel treatment to restore synaptic function following methamphetamine administration.

Cacna2d2 is an hnRNP H target of the striatal mRNA targetome and regulates methamphetamine behavior