Genetic Susceptibility and Epigenetic Modulation in Levodopa Induced Dyskinesia

 描述(由申请人提供)：此申请是为了在大卫·菲格的博士/博士培训的实验室阶段为他提供F31支持。该提案的科学重点是确定遗传影响在左旋多巴诱导的运动障碍(LID)的易感性和维持性中的作用，LID是左旋多巴治疗帕金森病的重要不良反应。LID很常见，50%的PD患者在治疗5年后发生LID。众所周知，LID具有显著的个体间变异性，尽管包括年龄和疾病严重程度在内的各种因素对LID的诱导有已知的影响，但它们无法完全解释这种观察到的变异性。使用在其他药物遗传学研究中取得成功但以前未用于帕金森病的“极端不协调”研究设计，我们将在患有帕金森病和LID的人类患者中寻找LID的重要遗传修饰物。下一步，我们将确定DNA甲基化的变化是否是LID中背侧纹状体细胞记忆长期变化的基础。使用LID的大鼠模型，我们将进行减少代表性的亚硫酸氢盐测序，以具体观察纹状体细胞敏化后CpG岛甲基化的变化。使用甲基化DNA免疫沉淀，我们将评估与LID相关的候选基因，并已知在大脑中经历动态DNA甲基化。我们还将从测序数据中验证已识别的甲基标记。DNA甲基化的重要性将通过在诱导LID的大鼠模型中阻断DNA甲基转移酶来进一步评估，以证明LID的形成是否需要DNA甲基化。大卫·菲格的拟议培训计划由项目导师大卫·斯坦德尔特博士以及合作者尼塔·林迪博士和理查德·迈尔斯博士赞助。培训计划的总体目标是为PI作为内科科学家的成功职业生涯奠定坚实的基础。通过提供一个以临床和实验室为基础的项目，同时专注于以疾病为导向的过程，是任何有抱负的内科科学家的理想培训环境。随着分子信息成为治疗决策中护理标准的一部分，收集和解释这些遗传影响的能力将是至关重要的。内科科学家通过对科学和医学的双重理解，将对这一过程起到至关重要的作用。通过向David提供了解遗传学对神经系统疾病的临床影响的技能，将使他能够扩展我们对分子过程的理解和应用，以实现患者个性化治疗。培训计划中包括的经验有助于PI：1)获得整合神经生物学和生物信息学的各种技术的能力；2)与其他科学家合作；3)发展假设驱动的研究；4)以书面和口头形式展示数据；5)有效地将研究与临床相结合；以及6)负责任地进行研究。

 DESCRIPTION (provided by applicant): This application is for F31 support of David Figge during the laboratory phase of his MD/PhD training. The scientific focus of the proposal is to determine the contribution of genetic influences in the susceptibility and maintenance of levodopa induced dyskinesia (LID), an important adverse effect of levodopa treatment for Parkinson's disease. LID is common, developing in 50% of PD patients after 5 years of treatment. LID is known to have significant inter-individual variability and although a variety of factors, including age and severity of disease, have known effects on LID induction they are unable to fully explain this observed variability. Using a "extreme discordant" study design which has been successful in other pharmacogenetic studies, but not previously employed in PD, we will search for important genetic modifiers of LID in human patients with PD and LID. Next, we will determine whether alterations in DNA methylation underlie the long standing changes in cellular memory of the dorsal striatum seen in LID. Using a rat model of LID, we will conduct reduced representation bisulfite sequencing to specifically look at changes in CpG island methylation following cellular sensitization in the striatum. Using methylated DNA immunoprecipitation we will assess candidate genes that are relevant in LID and known to undergo dynamic DNA methylation in the brain. We will also verify identified methyl marks from our sequencing data. The importance of DNA methylation will be further assessed by blocking DNA methyltransferases during the induction of LID in rat models to demonstrate if DNA methylation is required for LID formation. The proposed training plan for David Figge is sponsored by project mentor, Dr. David Standaert, and collaborators, Dr. Nita Limdi and Dr. Richard Myers. The overall goal of the training plan is to provide the PI with a solid foundation for a successful career as a physician scientist. By providing a project that is based both in the clinic and laboratory, while focused on a disease oriented process, is the ideal training environment for any aspiring physician scientist. As molecular information becomes a part of the standard of care in treatment decisions, the ability to gather and interpret these genetic influences will be of pivotal importance. Physician scientists will be vital to this process throug their dual understanding of science and medicine. By providing David with the skills to understand the clinical influence of genetics on neurological disease will enable him to expand our understanding and application of molecular processes to personalize patient treatments. Included in the training plan are experiences that help the PI: 1) gain competence in a variety of techniques integrating neurobiology and bioinformatics, 2) collaborate with other scientists, 3) develop hypothesis-driven research, 4) present data in a written and oral format, 5) effectively integrate research with clinic, and 6) responsibly conduct research.