Genomic Analyses for Elucidating Novel Targets for Symptoms Management

大多数症状是由多因素引起的，遗传和环境因素都会导致个体差异。在生物学假说的基础上进行了候选基因研究，以确定疼痛等复杂特征的相关遗传变异。然而，复杂的致病因素和涉及不同症状的数千个分子，使得很难检测到一个独特的疾病易感性个体的负责任的基因变异。单个基因中常见的变异不太可能主要作用于复杂的特征；相反，每个基因的贡献似乎很微妙，作用于多个生物途径中的一个，使其信号很难被检测到。 疾病知识的迅速增加以及应用强大和高容量技术的能力的共同影响，提高了人们对用于各种症状管理的更有效和更安全的药物的期望。为症状管理开发新的治疗策略也严重依赖于识别新的靶分子和确定特定类型疾病的表型。因此，这个项目的第一步是确定实验和临床表型的特征。 最近，我们启动了多个使用下一代测序(NGS)技术的项目，该技术允许我们执行完整和/或有针对性的基因组测序。将继续研究遗传和表观遗传因素在各种疾病和/或条件下的症状中的作用。例如，在急性疼痛、轻度创伤性脑损伤和创伤后应激障碍等神经系统疾病中，我们正在使用基因分型、基因和蛋白质表达以及患者报告的结果，以更好地了解这些因素与包括炎症级联在内的众多生物/机制途径之间的相互作用。从战区返回的士兵的基因表达谱在多次会议(CNRM和ASN会议)上提交，并已提交发表。两项表观遗传学研究使用DNA甲基化、染色质免疫沉淀和全基因组规模测序，分别来自平民和军事群体。运动相关脑震荡患者的表观遗传学变化将在ASHG会议上公布。第二项军队创伤后应激障碍患者的表观遗传学研究完成了样本测序，数据正在分析中。利用定向下一代测序技术，我们还有两个正在进行的微生物组项目，以表征来自不同疾病状态的人体和/或动物体内的微生物种群。使用新一代半导体测序仪，对肠易激综合征动物模型和人类患者的微生物组进行了广泛的16S rRNA区域测序。研究结果在科学会议上发表，并提交发表。最近启动了另一个针对再生障碍性贫血患者口腔微生物组的微生物组项目。到目前为止，初步数据分析已经完成，其结果将在机构间反兴奋剂机构会议上提出。将对更多的样本进行排序和分析。主要是这两个项目都发现，在特定的疾病条件下，微生物组的组成会发生变化。为下一代测序仪产生的高通量大数据开发新的分析方法是这些项目的另一个目标。 根据这些结果以及神经系统疾病中多个途径的生物学知识，我们将能够在个体水平上提出这些疾病的分子遗传机制。最后，我们可以建议进行综合基因组分析来开发新药，并基于个人遗传信息对其进行测试。

Most symptoms are multifactorial in origin with both genetic and environmental factors contributing to individual variations. Candidate gene studies on the basis of biological hypotheses have been performed to identify relevant genetic variation in complex traits such as pain. However, the complicated mesh of contributing factors and the thousands of molecules involved in different symptoms makes it difficult to detect responsible genetic variations for an individuals unique susceptibility to disorders. It is unlikely that common variations in a single gene act dominantly on complex traits; rather, the contribution of each gene seems to be subtle, acting on one of multiple biological pathways, making its signal difficult to detect. The combined impact of the rapid increase in knowledge of diseases and the ability to apply powerful and high capacity technology has raised expectations for more effective and safer medicines for various types of symptoms management. Developing new treatment strategies for symptoms management is also critically dependent on identifying new target molecules and defining phenotypes for specific types of disorders. Therefore the first step of this project has been to define the characteristics of experimental and clinical phenotypes. Recently, we launched multiple projects with next generation sequencing (NGS) technology, which allows us to perform entire and/or targeted genome sequencing. The role of genetic and epigenetic factors on symptoms in various disorders and/or conditions will continue to be studied. For example, in neurological disorders such as acute pain, mild traumatic brain injury and PTSD, we are using genotyping, gene and protein expression, and patient reported outcomes to better understand the reciprocal interplay between these factors and the numerous biologic/mechanistic pathways including the inflammatory cascade. Gene expression profiles from the soldiers back from war zone were presented at the multiple meetings (CNRM and ASN meeting) and have been submitted for publication. Two epigenetic studies using DNA methylation chromatin immunoprecipitation followed by whole genome scale sequencing were conducted from civilian and military groups. Epigenetic changes in the sports related concussion patients will be presented at the ASHG meeting. The second epigenetic study in military PTSD patients finished sequencing of samples and the data are under the analysis. Using the targeted next generation sequencing technology, we also have 2 on-going microbiome projects to characterize population of microorganisms in human and/or animal body from different disease status. Using a next generation semiconductor sequencer, microbiome was analyzed with extensive sequencing of 16s rRNA region in an animal model of irritable bowel syndrome and human patients. The results were presented at the scientific meeting and submitted for publication. Another microbiome project was launched recently for oral microbiome from the aplastic anemia patients. So far, preliminary data analysis was finished and its result will be presented at the IADR meeting. Additional samples will be sequenced and analyzed. Mainly both projects found that the microbiome composition is altered in specific disease conditions. Developing new analyzing methods for the high throughput big data generated by the next generation sequencers is another goal of these projects. From these results along with biological knowledge of multiple pathways in neurological disorders, we will be able to suggest molecular-genetic mechanisms of those diseases at the level of the individual. Finally, we can suggest integrative genomic analysis to develop new drugs and test them based on individual genetic information.