Expression genetics of pharmacogenes

药物基因的表达遗传学

• 批准号: 10376336
• 负责人: Danxin Wang
• 金额: $ 36.7万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-01 至 2026-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10376336
• 关键词: Adopted; Affect; Amino Acid Sequence; Architecture; Bioinformatics; Biological Markers; CYP2D6 gene; CYP3A4 gene; CYP7A1 gene; Clinical; Collaborations; Complex; Cytochrome P450; Enzymes; Epigenetic Process; Estrogen Receptor alpha; Funding; Gene Cluster; Gene Expression; Genes; Genetic; Genetic Epistasis; Genetic Markers; Genetic Variation; Genomics; Goals; Hepatocyte; Human; Knock-out; Ligands; Liver; Methodology; National Institute of General Medical Sciences; Outcome; Persons; Pharmaceutical Preparations; Pharmacogenetics; Pharmacotherapy; Positioning Attribute; Proteomics; Research; Sampling; Testing; Untranslated RNA; Variant; base; biomarker discovery; clinical biomarkers; clinical translation; clinically significant; cohort; design; drug efficacy; drug metabolism; enzyme activity; genetic variant; genome wide association study; induced pluripotent stem cell; innovation; insight; mRNA Expression; new technology; non-genetic; precision drugs; programs; protein expression; side effect; success

PROJECT SUMMARY The overarching goal of my research program is to gain a comprehensive understanding of the variability in drug metabolism to make drug therapies more effective and less toxic. The cytochrome P450 drug- metabolizing enzymes (dmCYPs) are the main enzymes that metabolize nearly 70% of the drugs currently used, which are highly variable from person to person, necessitating personalized drug therapy. Genetic factors significantly affect the drug metabolism, and the variants of some dmCYPs are used as clinical biomarkers to predict the metabolizer status. However, the underlying principles of their variable expression/activity are far from clear. Variants that result in altered amino acid sequences have been studied extensively, but they fail to account for all the differences observed. On the other hand, technical challenges have thus far limited the studies on factors that affect mRNA and protein expression of dmCYPs. Most dmCYPs are multi-gene clusters with complex genomic architecture and diverse genetic variations. With the support of NIGMS funding, we have adopted innovative methodologies and dissected the complex genomic architecture of multiple gene clusters (e. g., CYP3A) and identified hidden cis-acting regulatory variants that affect gene expression (e. g., 3A4, 3A5 and 3A43). Our results demonstrate that a single variant can have opposing effects on two genes in the same locus (e. g, 3A4, and 3A43), while more than one variant can regulate a single gene in the same or opposite direction (e. g., CYP7A1 and CYP2D6) indicating complex domain-domain interactions within the gene cluster and epistasis between variants. Such complexity usually escapes genome wide association study based discovery and warrants a dedicated approach proposed here. With the success of dissecting the CYP3A cluster, we now plan to expand and analyze the other dmCYP gene clusters over the next five years. Furthermore, we have recently identified ligand-free estrogen receptor alpha (ESR1) as a master regulator of CYP3A4 and many other dmCYP genes. Genetic, epigenetic, and non-genetic factors that affect ESR1-centered regulatory network will likely affect dmCYP gene expression and enzyme activity via trans-acting mechanisms. We plan to use ESR1-knockout hepatocytes differentiated from iPSC to characterize ESR1-centered regulatory network and to identify factors regulating ESR1 and dmCYPs expression. The identified functional cis and trans-acting genetic variants and epigenetic factors, including non-coding RNAs, will be tested in human liver samples for their impact on mRNA and protein expression of dmCYPs. The clinical significance of the identified variants will be tested in clinical cohort via collaboration. With my track record of pharmacogenetics biomarker discovery, insights gained from studying complex multi-gene clusters, availability of novel technologies, and support from collaborators with expertise in bioinformatics, proteomics and pharmacogenetics implementation, I am well positioned to undertake the proposed studies. The outcome will have immediate clinical translation leading to more accurate biomarkers for guiding personalized drug therapy.

项目摘要 我研究计划的首要目标是全面了解 药物代谢，使药物治疗更有效，毒性更小。细胞色素P450药物- 代谢酶（dmCYP）是目前代谢近70%药物的主要酶 使用，这是高度可变的人与人之间，需要个性化的药物治疗。遗传 药物代谢受多种因素的影响，一些dmCYP的变异体被用作临床药物代谢的候选基因。 生物标志物来预测代谢状态。然而，其变量的基本原理 表达/活动还远不清楚。已经研究了导致氨基酸序列改变的变体 广泛，但他们没有考虑到所有观察到的差异。另一方面，技术挑战 迄今为止，限制了对影响dmCYP的mRNA和蛋白质表达的因素的研究。最 dmCYP是具有复杂基因组结构和多样遗传变异的多基因簇。与 在NIGMS资金的支持下，我们采用了创新的方法， 多个基因簇的基因组结构（例如，例如，在一个实施例中，CYP 3A），并确定了隐藏的顺式作用调节 影响基因表达的变体（e.例如，在一个实施例中，3A 4、3A 5和3A 43）。我们的研究结果表明，一个单一的变量， 可以对同一基因座上的两个基因产生相反的影响（例如，g，3A 4和3A 43），而多于一个变体 可以在相同或相反的方向上调节单个基因（例如，例如，在一个实施例中，CYP 7A 1和CYP 2D 6），表明 基因簇内复杂的结构域相互作用和变异体之间的上位性。等 复杂性通常逃脱了基于全基因组关联研究的发现， 这里提出的方法。随着对CYP 3A簇的成功解剖，我们现在计划扩大和 在接下来的五年里分析其他的dmR基因簇。此外，我们最近发现， 无配体雌激素受体α（ESR 1）作为CYP 3A 4和许多其他dmR基因的主要调节因子。 影响以ESR 1为中心的调控网络的遗传、表观遗传和非遗传因素可能会影响 通过反式作用机制调节dmR基因表达和酶活性。我们计划使用ESR 1敲除 从iPSC分化的肝细胞表征以ESR 1为中心的调控网络并鉴定因子 调节ESR 1和dmCYP的表达。鉴定的功能性顺式和反式作用遗传变体和 表观遗传因素，包括非编码RNA，将在人类肝脏样本中测试其对 dmCYP的mRNA和蛋白表达。将检测已识别变体的临床意义 在临床队列研究中。根据我在药物遗传学生物标志物发现方面的记录， 从研究复杂的多基因簇，新技术的可用性，以及来自 在生物信息学，蛋白质组学和药物遗传学实施的专业知识的合作者，我很好 准备进行拟议的研究。结果将立即临床转化， 更准确的生物标志物，用于指导个性化药物治疗。