Translating Genetic Risk Factors to Therapies: From Big Data to Druggable Targets

项目总结 全基因组关联研究提供了对不明显疾病的潜在病因的洞察。 仅通过临床评估或病理生理学。与复杂的、通常是难治的遗传变异有关 阿尔茨海默病(AD)和多发性硬化症(MS)等疾病可能是下一代的关键 治疗方法的选择。利用遗传数据和当前的护理标准和已知的病理生理学可以 为机械学研究和新的药物靶点提供了强有力的前提。我目前的工作是研究分子 史蒂夫·埃斯特斯博士指导下的阿尔茨海默病患者CD33的遗传学研究。CD33通常作用于抑制大脑中的小胶质细胞激活， 抑制淀粉样蛋白清除。我们正在研究AD保护性单核苷酸多态性是如何 CD33中的(SNP)调节蛋白质，从而调节细胞功能。这种SNP导致了另一种选择的增加 根据我们最近的基因数据，CD33蛋白亚型可能促进而不是抑制小胶质细胞 激活。我将在这个奖项的F99阶段学习新的技术技能，我将在转换过程中使用这些技能 专注于MS并进入K00阶段，以获得其他强大的技术和模型，包括 研究患者样本、多能干细胞、单细胞测序技术和动物模型。 在具体目标1中，我详细介绍了我在分子生物学和遗传概念方面的培训如何使我能够 构思独立的假设，进行复杂的实验。我在分子方面的博士工作 CD33的遗传学及其与AD风险降低的关联提供了GWAS解释方面的培训， 定量聚合酶链式反应，免疫分析，如免疫印迹和免疫共沉淀，以及基因技术 包括细胞培养中的转基因和基因组编辑策略。在具体目标2中，我将继续发展 作为一名科学家，完成了我的博士工作，进行了越来越复杂的研究，包括高分辨率 共聚焦成像和亚细胞定位，测量时间和剂量依赖性的蛋白质磷酸化 原位、基因表达阵列和功能分析，包括体外吞噬作用。我也会继续发展 我的专业技能，如口头和书面沟通，网络，和指导。在具体目标3中，我 我将延长我的博士培训，以包括对人类组织样本和I女士的老鼠模型的研究 我还没有确定具体的博士后导师，但我理想的导师将有指导经验 人类受试者研究，使用多发性硬化症的老鼠模型，并有良好的培训研究员记录 成为终身教职的教师。我会找一个指导团队来指导我的技术和专业 这一阶段的发展。我将利用我目前在分子遗传学方面的培训来识别与多发性硬化症相关的 功能性SNPs，我正在进行的培训，以确定这些SNPs背后的机制在蛋白质和 细胞内信号水平，以及我未来用小鼠模型和人类受试者建立的高- 影响，翻译职业，结合遗传学、临床和病理学的药理学研究结果 神经免疫疾病的突破性进展。

PROJECT SUMMARY Genome-wide association studies (GWAS) provide insight to underlying etiologies of disease not obvious through clinical evaluation or pathophysiology alone. Genetic variants associated with complex, often refractory diseases such as Alzheimer’s Disease (AD) and Multiple Sclerosis (MS) may hold the key for the next generation of treatment options. Leveraging genetic data with current standards of care and known pathophysiology can provide a strong premise for mechanistic studies and novel drug targets. My current work studies the molecular genetics of CD33 in AD under Dr. Steve Estus. CD33 normally acts to inhibit microglial activation in the brain, suppressing amyloid clearance. We are investigating how the AD-protective single nucleotide polymorphism (SNP) in CD33 modulates protein, and thereby cellular, function. This SNP leads to an increase in an alternative CD33 protein isoform which, based on our recent genetic data, may promote—rather than suppress—microglial activation. I will learn new technical skills during the F99 phase of this award, and I will use these skills as I switch focus to MS and progress into the K00 phase to acquire additional, powerful techniques and models including work with patient samples, pluripotent stem cells, single-cell sequencing technologies, and animal models. In Specific Aim 1, I detail how my training in molecular biology and genetic concepts has allowed me to conceive independent hypotheses and carry out complex experiments. My doctoral work on the molecular genetics of CD33 and its association with reduced AD risk has provided training in GWAS interpretation, quantitative PCR, immunoassays such as Western blotting and co-immunoprecipitation, and genetic techniques including transfection and genome editing strategies in cell culture. In Specific Aim 2, I will continue to develop as a scientist and finish my doctoral work, carrying out increasingly complex studies to include high-resolution confocal imaging and subcellular localization, measuring time- and dose-dependent protein phosphorylation in situ, gene expression arrays, and functional assays including phagocytosis in vitro. I will also continue developing my professional skills such as oral and written communication, networking, and mentorship. In Specific Aim 3, I will extend my doctoral training to include work with human tissue samples and mouse models of MS. I have not yet identified a specific postdoctoral mentor, but my ideal mentor will have experience conducting human subjects research, using mouse models of MS, and have a strong track record of training fellows to become tenure-track faculty. I will identify a mentorship team to guide my technical and professional development during this phase. I will leverage my current training in molecular genetics to identify MS-associated functional SNPs, my training-in-progress to identify the mechanism behind these SNPs at the protein and intracellular signaling levels, and my future training with murine models and human subjects to establish a high- impact, translational career, combining genetic, clinical, and pathology findings for pharmacological breakthroughs in neuroimmune diseases.