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

将遗传风险因素转化为治疗方法：从大数据到可药物靶点

基本信息

批准号：
10318416
负责人：
Benjamin C. Shaw
金额：
$ 3.89万
依托单位：
UNIVERSITY OF KENTUCKY
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-07-01 至 2022-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10318416
关键词：
Address Affect Alternative Splicing Alzheimer&apos s Disease Alzheimer&apos s disease risk American Amyloid Amyloid beta-Protein Animal Model Antigen-Antibody Complex Award Big Data Biological Assay Brain CRISPR/Cas technology Caring Cell Culture Techniques Cell Line Cell Surface Receptors Cell surface Cells Cellular biology Clinical Pathology Clinical Trials Co-Immunoprecipitations Communication Complex Confocal Microscopy Data Data Analyses Development Disease Dose Drug Targeting Educational workshop Electronic Health Record Engineering Enzyme-Linked Immunosorbent Assay Etiology Exons Faculty Functional disorder Future Gene Chips Gene Expression Genetic Genetic Techniques Goals Heritability Human Human Subject Research Immune Immunoassay Impairment In Situ In Vitro Individual Institution Knowledge Learning Ligand Binding Domain Manuscripts Measures Mentors Mentorship Microglia Microscopy Modeling Molecular Biology Molecular Genetics Molecular and Cellular Biology Multiple Sclerosis Myeloid Cells Neurodegenerative Disorders Neuroimmune Neurosciences Oral PTPN6 gene Patients Peptides Phagocytosis Pharmacology Phase Phosphoric Monoester Hydrolases Phosphorylation Phosphotransferases Pluripotent Stem Cells Positioning Attribute Protein Isoforms Proteins Proto-Oncogene Proteins c-akt Quality of life RNA Splicing Refractory Disease Research Research Personnel Resolution Risk Factors Role Sampling Schools Scientist Signal Transduction Single Nucleotide Polymorphism Small Interfering RNA Societies System TREM2 gene Technical Expertise Techniques Technology Therapeutic Time Tissue Sample Training Transfection Translating Tyrosine Variant Viral Western Blotting Work abeta toxicity base brain cell career confocal imaging data mining druggable target effective therapy experience experimental study gain of function genetic risk factor genetic variant genome editing genome wide association study graduate student human data human disease human subject human tissue hyperphosphorylated tau immunoregulation improved induced pluripotent stem cell inhibitor/antagonist insight meetings monocyte mouse model nano-string neurotoxicity new therapeutic target next generation novel overexpression post-doctoral training receptor research clinical testing sample collection single cell sequencing single-cell RNA sequencing skills symposium tau Proteins tenure track therapeutic target undergraduate student

项目摘要

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.

项目摘要全基因组关联研究（GWAS）为疾病的潜在病因提供了见解仅通过临床评估或病理生理学。与复合物相关的遗传变异，通常是难治性的阿尔茨海默氏病（AD）和多发性硬化症（MS）等疾病可能是下一代的关键治疗选择。利用当前护理标准和已知病理生理学标准的遗传数据可以为机械研究和新型药物靶标提供了有力的前提。我目前的工作研究分子史蒂夫·埃斯特图斯（Steve Estus）博士在AD中的CD33的遗传学。 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保护单一核苷酸多态性（SNP）在CD33中调节蛋白质，从而调节细胞功能。该SNP导致替代方案增加 CD33蛋白质同工型基于我们最近的遗传数据，可能会促进（抑制）microglial 激活。在此奖项的F99阶段，我将学习新的技术技能，我将在切换时使用这些技能专注于MS并进入K00阶段，以获取其他功能强大的技术和模型，包括与患者样品，多能干细胞，单细胞测序技术和动物模型一起工作。在特定目标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想独立的假设并进行复杂的实验。我在分子上的博士学位 CD33的遗传学及其与降低的AD风险的关联提供了GWAS解释的培训，定量PCR，诸如蛋白质印迹和共免疫沉淀等免疫测定以及遗传技术包括细胞培养中的转化和基因组编辑策略。在特定目标2中，我将继续发展作为科学家并完成我的博士工作，进行越来越复杂的研究以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。我也将继续发展我的专业技能，例如口头和书面交流，网络和心态。在特定的目标3中，我将扩展我的博士培训，包括与MS的人体组织样本和小鼠模型的工作。我尚未确定特定的博士后心理，但是我理想的心理会有经验人类受试者使用MS的鼠标模型进行研究，并具有训练研究员的良好记录成为终身教师。我将确定一个心态团队来指导我的技术和专业人士在此阶段的发展。我将利用目前在分子遗传学上的培训来识别与MS相关的功能性SNP，我进行的训练，以识别蛋白质上这些SNP背后的机制细胞内信号传导水平，以及我对鼠模型和人类受试者的未来培训，以建立高影响，翻译职业，结合了药物的遗传，临床和病理发现神经免疫性疾病的突破。