Defining the role of developmental context in oncohistone-driven gliomagenesis

项目摘要 这项提案的首要目标是定义表观遗传学和发育背景之间的相互作用。 驱动儿童高级别胶质瘤(PHGG)，由组蛋白突变定义的普遍致命肿瘤，以便 找出更好的针对性治疗方法。这项工作利用了一种创新的基于大脑器官的模型，该模型允许 癌组蛋白突变体的并列比较和癌基因表达的动态调节 多维的人类环境。利用这一创新模型，格雷厄姆博士将使用尖端的单细胞 侧写技术和临床前胶质瘤建模以解决目前阻碍 这些患者的治疗进展。在目标1中，染色质分析和单细胞RNA测序将 H3.3K27M和H3.3G34V两种不同的肿瘤组蛋白在神经发育过程中的作用 发展。在目标2中，突变的时间和顺序将被操纵以评估细胞的影响 以及肿瘤表型的突变背景。最后，在目标3中，正交遗传“拯救”和药理学 抑制方法将被用来询问这些突变在肿瘤维持中的作用。 这项工作，以及格雷厄姆博士的职业目标，都与NINDS的使命非常一致，即寻求基本的 关于大脑的知识并将这些知识转化为临床影响。通过拟议的研究 以及随之而来的职业发展计划，格雷厄姆博士将获得表观遗传学方面的基本培训， 生物信息学和临床前胶质瘤研究，是她目前技能的关键空白。格雷厄姆博士是一位 纪念斯隆·凯特琳癌症中心英戈·梅林霍夫医生指导下的神经肿瘤学 (MSK)。梅林霍夫博士是翻译性胶质瘤研究的领导者，在指导方面有很好的记录 实习生走向独立。他和格雷厄姆博士组建了一个特殊的咨询委员会， 染色质生物学、翻译肿瘤学和生物信息学分析方面的专业知识：克里斯蒂安·海林博士、罗斯博士 莱文和尼古拉斯·索奇博士。MSK为培养初出茅庐的职业提供了良好的环境 生物医学研究，拥有无与伦比的资源、支持和合作机会。vt.在.的基础上 完成拟议的工作后，格雷厄姆博士将处于理想的位置，并具有独特的职业资格 阐明神经谱系表观遗传调控的基本方面的独立研究人员 致力于并应用她的发现来解决恶性胶质瘤治疗中未得到满足的需求。

Project Summary The overarching goal of this proposal is to define the interplay of epigenetics and developmental context in driving pediatric high-grade gliomas (pHGG), universally lethal tumors defined by histone mutations, so as to identify better targeted treatments. This work utilizes an innovative cerebral organoid-based model that allows side-by-side comparison of oncohistone mutants and dynamic modulation of oncogene expression in a three- dimensional human context. Leveraging this innovative model, Dr. Graham will use cutting-edge single cell profiling techniques and preclinical glioma modeling to address unanswered questions that currently hamper therapeutic development for these patients. In Aim 1, chromatin profiling and single cell RNA sequencing will be used to investigate the distinct effects of two different oncohistones (H3.3K27M and H3.3G34V) during neural development. In Aim 2, the timing and order of mutations will be manipulated to evaluate the impact of cellular and mutational context on tumor phenotype. Finally, in Aim 3, orthogonal genetic “rescue” and pharmacologic inhibition approaches will be used to interrogate the role of these mutations in tumor maintenance. This work, as well as Dr. Graham's career goal, is well-aligned with the NINDS mission to seek basic knowledge about the brain and to translate that knowledge into clinical impact. Through the proposed studies and the accompanying career development plan, Dr. Graham will gain essential training in epigenetics, bioinformatics and preclinical glioma studies, critical gaps in her current skillset. Dr. Graham is an Instructor of Neuro-Oncology under the mentorship of Dr. Ingo Mellinghoff at Memorial Sloan Kettering Cancer Center (MSK). Dr. Mellinghoff is a leader in translational glioma research with a strong track record of mentoring trainees to independence. He and Dr. Graham have assembled an exceptional Advisory Committee with expertise in chromatin biology, translational oncology, and bioinformatic analyses: Dr. Kristian Helin, Dr. Ross Levine and Dr. Nicholas Socci. MSK provides an outstanding environment for cultivating budding careers in biomedical research, with unparalleled resources, support, and opportunities for collaboration. Upon completion of the proposed work, Dr. Graham will be ideally positioned and uniquely qualified for a career as an independent investigator elucidating fundamental aspects of epigenetic regulation in neural lineage commitment and applying her findings to address the unmet needs in the treatment of malignant glioma.