BLRD Research Career Scientist Award Application

项目摘要/摘要 弗里曼博士这项研究的首要目标是了解 基因组/表观基因组有助于衰老，以及治疗如何潜在地针对这些变化来维持 延缓衰老，延缓/预防与年龄相关的疾病，并改善健康和 老年人的生活质量。表观遗传失调是衰老的一个标志，但这到底是如何导致 细胞功能障碍和疾病尚不清楚。为了阐明这一贡献，弗里曼博士的退伍军人事务部研究已经 重点研究了星形胶质细胞、小胶质细胞和海马神经元随年龄增长的表观基因组调控。vbl.使用 新颖的细胞分离和先进的测序方法，两者的细胞类型特定碱基分辨率图 在CG和非CG环境中正在产生甲基化(MC)和羟甲基化(HMC)。与配对 RNA表达数据，生物信息学技术正被用来理解基因表达的调节 随着年龄的增长。这种类型的碱基特异性、全基因组和细胞类型特异性研究还没有在 老龄化研究，并可能提供迄今为止关于表观基因组变化的最深入的观点。 此外，为了证明防止与年龄相关的表观基因组变化，正在使用卡路里限制。在……里面 总之，这些研究不仅将增加对衰老的神经表观组学的理解，而且还将提供 使用表观基因组编辑的未来干预性研究的目标。重要的是，这些研究检查了这两个男性 和女性提供对性知识的洞察力，因为他之前的研究表明，大多数年龄- 相关的表观基因组变化是性别分化的。 弗里曼博士也是NIH三个项目和一个基金会资助项目的PI或联合PI。这些研究扩展了 他通过使用新的转基因小鼠模型和大脑抗衰老干预在衰老方面的VA研究， 视网膜和胃肠系统。这些由他的实验室开发的模型允许时间控制的、细胞类型的特定标记 用于表观基因组分析的DNA和用于表达分析的RNA 需要对细胞进行分选。在大脑和视网膜中，异时血浆转移，其中来自幼鼠的血浆 用在老年小鼠身上以恢复活力，正在被用来确定与年龄相关的表观基因组变化是否可以 在它们发生后颠倒。同时，使用蛋白质组学方法，DNMT和Tet相互作用的蛋白质 它们被确定为针对特定基因组区域进行表观基因组改变。 更广泛地说，弗里曼博士还与OKC VAMC的其他调查人员进行了广泛的合作。在……里面 这些由退伍军人事务部和美国国立卫生研究院资助的研究，弗里曼博士正在研究肌肉衰老的细胞类型特定机制， 大脑、卵巢、关节和结肠。除了为这些研究带来分子和生化专业知识外， 衰老表观遗传学的共同线索允许跨器官系统进行比较和对比。 这些合作与他作为最近更新的一个基因组科学核心的主任的工作相匹配 P30 NIA中心赠款：俄克拉荷马州内森休克衰老中心。这个核心提供了表观基因组学和 对全国各地的老年研究人员进行转录分析和线粒体基因组分析。 此外，弗里曼博士还领导了许多跨机构(OMRF、OUHSC)计划，重点关注学员和教职员工 发展，包括培训学生和博士后(F-Troop)和初级教员(NeuroGrant Seeking) 成功地准备和提交了赠款。他还与OKC VAMC的领导人合作，帮助 招募新的、非常成功的调查人员到退伍军人制度，其中一人获得了优秀奖 又有三名调查人员目前正在审查优秀的申请。

Project Summary/Abstract The overarching goal of Dr. Freeman’s research has been to understand how alterations of the genome/epigenome contribute to aging and how therapies could potentially target these changes to maintain a ‘youthful’ epigenome that retards aging, delays/prevents age-related diseases, and improves the health and quality of life of the elderly. Epigenetic dysregulation is a hallmark of aging, but exactly how this contributes to cellular dysfunction and disease is not known. To elucidate this contribution, Dr. Freeman’s VA research has focused on epigenomic regulation in astrocytes, microglia, and neurons in the hippocampus with aging. Using novel cell isolation and advanced sequencing approaches, cell-type specific base resolution maps of both methylation (mC) and hydroxymethylation (hmC) in CG and non-CG contexts are being generated. With paired RNA expression data, bioinformatic techniques are being used to understand the regulation of gene expression with aging. This type of base-specific, genome-wide, and cell-type specific studies have not been performed in aging research and will potentially offer the most in-depth view of epigenomic changes with aging to date. Further, to demonstrate the prevention of age-related epigenomic changes, caloric restriction is being used. In total, these studies will not only increase the understanding of the neuroepigenomics of aging but also provide targets for future interventional studies using epigenome editing. Importantly these studies examine both males and females to provide sex-informed insight as his prior studies have demonstrated that the majority of age- related epigenomic changes are sexually divergent. Dr. Freeman is also PI or co-PI on three NIH projects and one foundation funded project. These studies extend his VA research in aging through use of novel transgenic mouse models and anti-aging interventions in the brain, retina, and GI system. These models developed by his lab allow temporally controlled, cell-type specific tagging of nuclei and ribosomes to isolate DNA for epigenomic analysis and RNA for expression analysis without the need for cell sorting. In the brain and retina, heterochronic plasma transfer, in which plasma from young mice is administered to old mice for rejuvenation, is being used to determine if age-related epigenomic changes can be reversed after they have occurred. In parallel, using proteomic approaches, DNMT and TET interacting proteins are being identified that target them to specific genomic regions for epigenomic alteration. More broadly, Dr. Freeman also has an extensive collaborative portfolio with other OKC VAMC investigators. In these VA and NIH funded studies, Dr. Freeman is examining cell-type specific mechanisms of aging in muscle, brain, ovary, joints, and colon. In addition to bringing molecular and biochemical expertise to these studies, a common thread of epigenetics of aging allows comparisons and contrasts to be made across the organ systems. These collaborations are paired with his work as Director of the Genomics Sciences Core of a recently renewed P30 NIA Center grant: the Oklahoma Nathan Shock Aging Center. This core provides epigenomic and transcriptomic analyses as well as mitochondrial genomic assays for aging researchers across the country. Additionally, Dr. Freeman leads many cross-institution (OMRF, OUHSC) initiatives focused on trainee and faculty development, including training students and post-docs (F-Troop) and junior faculty (NeuroGrant Seekers) in successful preparation and submission of grants. He has also worked with leaders at the OKC VAMC to help recruit new, highly successful investigators to the VA system, one of which has received a MERIT award with three more investigators having MERIT applications currently under review.