The Immortality and Evolution of Adult Brain Tumors

项目摘要 科斯特洛实验室研究脑瘤中的细胞永生和进化，目的是将 我们对新疗法的发现。我在我们的学习中扮演着重要的角色，以理解和克服 由肿瘤进化驱动的基因组和表观基因组肿瘤内异质性，这是治疗的基础 失败。一个翻译目标是识别产生免疫原性新抗原的突变 在整个肿瘤中都存在，与Dr. 冈田。与一个跨学科的临床团队合作，我们开发了一种独特的、三维的完整肿瘤 抽样方法，其中我们从每个肿瘤中选择10个空间映射样本 神经外科医生最大限度地代表整个肿瘤。我们已经开始应用基因组， 从这些空间定位样本产生的转录和T细胞受体(TCR)测序数据I 鉴定肿瘤中存在的免疫原性新抗原及其同源TCR。我也是一名 对我们的肿瘤不朽研究做出了重大贡献。为了无限增殖，肿瘤细胞必须克服 正常的寿命极限在很大程度上是由端粒缩短决定的，端粒缩短是细胞分裂的结果 端粒酶活性缺失。80%的胶质母细胞瘤和许多其他癌症都能克服这一点 通过获得端粒酶启动子突变实现细胞永生的寿命障碍 逆转录酶(TERT)。我们发现，TERT启动子突变会激活正常的 转录因子GABP选择性募集沉默TERT基因与端粒酶活性 这通常不会对TERT进行监管。利用GABP的实验靶向，我们证明了TERT 在带有TERT启动子突变的细胞中选择性地降低表达，当与 化疗后，它会显著减少体内GBM的生长。目前，我正在研究一种新发现的 GABP亚单位表达的动态平衡调控及其与TERT调控的关系。要翻译这些内容 一种新疗法的机械学研究，科斯特洛博士与人共同创立了一家生物技术初创公司，该公司发现 具有药物性质的小分子，以突变依赖的方式降低TERT。除了……之外 为了实现这些翻译目标，我的职责包括：(1) 管理和监督实验室的基础设施和肿瘤样本采集；(2)技术和 为小组开发方法；(3)确保所有实验室研究在 符合法规要求；以及(4)培训整个实验室的新成员和个人 脑肿瘤中心。这个R50应用程序请求为这些正在进行的活动提供薪资支持 NCI资助的以下项目的科学目标：发现基因组效应器的三维空间方法 恶性转化过程中的免疫抑制(R01 CA244838)；脑瘤孢子P1-A TERT启动子突变型胶质瘤的新治疗靶点(2P50CA097257)

PROJECT ABSTRACT The Costello lab investigates cellular immortality and evolution in brain tumors with the goal of translating our discoveries into new therapies. I play an essential role in our studies to understand and overcome the genomic and epigenomic intra-tumor heterogeneity, driven by tumor evolution, that underlies therapeutic failures. One translational goal is to identify mutations that produce immunogenic neoantigens that are present throughout the whole tumor, to develop personalized immunotherapies in collaboration with Dr. Okada. Working with an interdisciplinary clinical team, we developed a unique, 3-dimensional whole tumor sampling approach in which we obtain 10 spatially mapped samples per tumor selected by the neurosurgeon to maximally represent the whole tumor. We have begun applying the genomic, transcriptomic and T cell receptor (TCR) sequencing data I produced from these spatially mapped samples to identify immunogenic neoantigens and their cognate TCR present throughout the tumor. I am also a significant contributor to our tumor immortality studies. To proliferate indefinitely, tumor cells must overcome the normal limits on lifespan dictated in large part by telomere shortening, a consequence of cell divisions in the absence of telomerase activity. Eighty percent of glioblastoma and many other cancers overcome this lifespan barrier to achieve cellular immortality by acquiring a mutation in the promoter of Telomerase Reverse Transcriptase (TERT). We discovered that the TERT promoter mutation activates the normally silent TERT gene and telomerase activity through selective recruitment of GABP, a transcription factor which does not normally regulate TERT. Using experimental targeting of GABP, we showed that TERT expression is reduced selectively in cells with the TERT promoter mutation, and when combined with chemotherapy, it dramatically reduces GBM growth in vivo. Currently, I am studying a newly discovered homeostatic control on GABP subunit expression and its relationship to TERT regulation. To translate these mechanistic studies into a new therapy, Dr. Costello co-founded a biotech startup which has discovered small molecules with drug-like properties that reduce TERT in a mutation dependent manner. In addition to performing bench research to address these translational goals, my responsibilities include: (1) management and oversight of the lab’s infrastructure and tumor sample collection; (2) technology and methods development for the group; (3) ensuring that all laboratory research is conducted safely in accordance with regulatory requirements; and (4) training new lab members and individuals throughout the Brain Tumor Center. This R50 application requests salary support for these ongoing activities to advance the scientific goals of the following NCI-funded projects: 3-D spatial approach to discover genomic effectors of immunosuppression during malignant transformation (R01 CA244838); The Brain Tumor SPORE P1 – A New Therapeutic Target for TERT Promoter Mutant Glioma (2P50CA097257).