High-Throughput Functional Genomics of Variants in Genes Linked to Substance Use Disorders

摘要 药物使用障碍相关基因变异的高通量功能基因组学研究 对物质使用障碍的遗传基础的理解在最近的过去有了显著的进步； 2000年初S参考人类序列的产生使基因组广谱关联研究成为可能 这反过来又导致了数以千计的具有统计学意义的遗传变异的鉴定 与物质使用和物质使用障碍有关。这些与临床相关的变异的数量 随着GWAS和基因组测序研究样本量的增加，继续按比例增长 药物使用障碍。模式生物研究也导致了对基因和途径的识别 与这些表型相关的基因。然而，这一领域的一个长期瓶颈是缺乏 对基因和调控区域中几乎所有(&gt；99%)这些变异进行实验测试，以确定 它们在功能上具有重要意义。确定遗传变异的功能效应对于获得 对物质使用障碍基础的分子理解，这种更深层次的理解将使 未来有针对性和合理设计的治疗干预措施的开发和测试。这位前锋- Think Application建议通过应用高度创新的高吞吐量解决这一差距 功能基因组学方法，包括大规模平行报告分析(MPRAS)和深度分析 突变扫描(DMS)技术，最近被用于研究罕见的孟德尔人 精神障碍，研究数千种与物质相关的非编码和编码变体 使用障碍。这些方法以前没有被应用于物质使用障碍的遗传学， 尽管它们在其他领域取得了重大进展，例如 神经发育和神经精神障碍。此外，这位早期调查人员的 多学科培训，包括医学/博士学位、儿科住院医师、博士后奖学金和四个 多年致力于K08早期职业发展的工作，专注于获得使用 功能基因组学技术，使他成为人类遗传学领域的一名冉冉升起的领导者。他的前科 改变范式和基于团队的成就包括建立外显子组测序测试 2015年自闭症儿童的管道，目前正在协调模式生物筛查中心 NIH资助的未诊断疾病网络，以及领导一个150万美元的机构资助的多 调查员功能基因组学项目，导致部署关键基础设施并提供10：1 后续赠款资金的投资回报。尽管这位才华横溢的调查员之前并没有 在物质使用障碍方面，他与该领域的世界专家劳拉·比鲁特博士建立了合作关系， 世卫组织已同意在这一研究项目中为他提供建议，并将他与 菲尔德。总而言之，这一极具影响力和极具创新性的提议来自一位特别合格的早期- 阶段性研究人员将大大促进对物质使用障碍遗传学的理解。

ABSTRACT High-throughput functional genomics of variants in genes linked to substance use disorders Understanding of the genetic basis of substance use disorders has advanced significantly in the recent past; the generation of a reference human sequence in the early 2000’s enabled Genome Wide Association Studies (GWAS), which in turn led to the identification of thousands of genetic variants that are statistically significantly associated with substance use and substance use disorders. The number of these clinically relevant variants continues to grow proportionally with the increase in sample sizes for GWAS and genome sequencing studies of substance use disorders. Model organism studies have also led to the identification of genes and pathways that are linked to these phenotypes. However, one persistent bottleneck in this field has been the lack of experimental testing of nearly all (>99%) of these variants in genes and regulatory regions to determine which are functionally significant. Determining the functional effects of genetic variants is crucial to acquiring a molecular understanding of the basis of substance use disorders, and this deeper understanding will enable future development and testing of targeted and rationally designed therapeutic interventions. This forward- thinking application proposes to address this gap by applying highly innovative high-throughput functional genomics approaches, including Massively Parallel Reporter Assays (MPRAs) and Deep Mutational Scan (DMS) techniques, that were recently deployed for the study of rare Mendelian disorders, to the study thousands of both noncoding and coding variants associated with substance use disorders. These approaches have not previously been applied to the genetics of substance use disorders, although they have enabled significant advancements in other fields, such as the genetics of neurodevelopmental and neuropsychiatric disorders. Furthermore, this early-stage investigator’s multidisciplinary training, including MD/PhD degrees, Pediatrics residency, a postdoctoral fellowship, and four years of dedicated K08 Early Career Development work that was focused on acquiring expertise in the use of functional genomics techniques, has established him as a rising leader in the field of human genetics. His prior paradigm-altering and team-based accomplishments include establishment of an exome sequencing testing pipeline for children with autism in 2015, ongoing coordination of the Model Organism Screening Center for the NIH-funded Undiagnosed Diseases Network, and leadership of a $1.5 million institutionally-funded, multi- investigator functional genomics project that resulted in deployment of critical infrastructure and delivered a 10:1 return on investment in subsequent grant funding. Although this talented investigator has not previously focused on substance use disorders, he has established a collaboration with a world expert in this field, Dr. Laura Bierut, who has agreed to advise him in this research program and connect him with other relevant investigators in the field. In sum, this very high impact and extremely innovative proposal from a particularly well qualified early- stage investigator will significantly advance the understanding of the genetics of substance use disorders.