Abstract The purpose of the NIDA Animal Genetics Program is to identify genetic, genomic, epigenetic variants, physiology and brain functions that contribute to addiction-like behaviors, related behavioral endophenotypes, and behavioral comorbidities to substance use disorder. During the past four years, our multidisciplinary and highly collaborative consortium has been identifying gene variants that are associated with increased vulnerability to compulsive-like cocaine use by performing the first GWAS using an advanced model of chronic intravenous cocaine self-administration in N/NIH heterogeneous stock (HS). We have also created the first preclinical cocaine biobank which enables researchers who do not have the resources to perform chronic intravenous self-administration or next-generation genome sequencing to perform advanced genetic, molecular, and cellular studies to further our understanding of the biological changes underlying addiction-like behaviors. While these efforts have been very successful in achieving the planned milestones, it has become clear that our project would benefit from an even larger sample size. In particular, increasing sample sizes lead to exponential rather than linear increase in the number of loci identified. Moreover, in the past four years there has been tremendous technological advances in behavioral and genetic analysis that can be leveraged to provide unprecedented access to identify the single nucleotide and structural variants that contribute to complex behavioral endophenotypes of high relevance to cocaine use-disorders. The first goal of this competing renewal is to double the sample size of the current GWAS to increase the number of gene variants identified and meet the demands of the Biobank. The second goal is to use high-throughput behavioral phenotyping using markerless pose estimation based on transfer learning with deep neural network to identify behavioral endophenotypes that can help predict and identify individuals with a resistant, mild, moderate, or severe phenotype of cocaine addiction-like behaviors. The third goal is to use methodological improvements of the genetic analysis, including the analysis of structural variants and tandem repeats, as well as enhanced integration with gene expression data. The fourth goal is to strengthen the cocaine biobank infrastructure. This project is likely to continue having a sustained and powerful impact on the field because it will provide an exponential increase in the number of genetic loci identified, eQTLs and PheWAS analysis related to addiction- like behavior; establish the first high-throughput behavioral motifs analysis of addiction-like behaviors using parallel video-recording and automated machine learning analysis; identify novel behavioral endophenotypes of vulnerability/resistance to addiction-like behaviors; and expand and improve the Cocaine Biobank offering and infrastructure.

Narrative This is a multidisciplinary, highly collaborative consortium that combines next-­generation sequencing with state-­of-­the-­art behavioral screening in a unique, genetically diverse, nonhuman animal model. The goals of this consortium are to identify gene variants that are associated with compulsive-like cocaine intake and behavioral endophenotypes using genome-­wide association studies (GWAS) in behaviorally characterized heterogeneous stock rats, and to strengthen the infrastructure of a newly created data/tissue repository (Cocaine Biobank) accessible to all researchers to study the cellular and molecular mechanisms related to cocaine use disorder. Results from these studies have the potential to have a sustained and powerful impact on the field of addiction because they will identify novel druggable targets, provide a comprehensive analysis of cocaine addiction-like behaviors in males and females, and increase the impact of a unique data/tissue repository that will facilitate follow-­up and replication studies.