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

摘要 物质使用障碍相关基因变异的高通量功能基因组学 近年来，对物质使用障碍的遗传基础的理解有了很大的进展; 在2000年代早期，一个参考人类序列的产生使得全基因组关联研究成为可能， （GWAS），这反过来又导致了数千种遗传变异的鉴定，这些变异在统计学上是显著的。 与物质使用和物质使用障碍有关。这些临床相关变异的数量 随着GWAS和基因组测序研究样本量的增加， 物质使用障碍模式生物的研究也导致了基因和途径的识别 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 实验测试几乎所有（>99%）这些基因和调控区的变异，以确定 这在功能上是重要的。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，这种更深入的理解将使 未来的发展和测试有针对性的和合理设计的治疗干预措施。这个前锋- Thinking Application建议通过应用高度创新的高通量技术来解决这一差距 功能基因组学方法，包括大规模平行报告基因测定（MPRAs）和深度 突变扫描（DMS）技术，最近部署用于研究罕见的孟德尔遗传 研究了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，如遗传学， 神经发育和神经精神障碍。此外，这个早期的调查员 多学科培训，包括医学博士/博士学位，儿科住院医师，博士后奖学金，和四个 多年专注于K 08早期职业发展工作，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位冉冉升起的领导者。他之前 范式改变和基于团队的成就包括建立外显子组测序测试 在2015年为自闭症儿童提供的管道中，正在进行的协调模式生物筛选中心， 美国国立卫生研究院资助的未诊断疾病网络，以及一个150万美元的机构资助的多个 调查员功能基因组学项目，导致部署关键基础设施，并交付了10：1的 投资回报率在随后的赠款资金。尽管这位才华横溢的调查员此前并没有专注于 在药物使用障碍方面，他与这一领域的世界专家Laura Bierut博士建立了合作关系， 他同意在这项研究计划中为他提供建议，并将他与其他相关调查人员联系起来。 领域总之，这一非常高的影响力和极具创新性的建议，从一个特别合格的早期- 阶段研究者将大大推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.