Integrative Approaches to Stroma-Directed Glioma Therapy

概述（由申请人提供）：总而言之，该提案侧重于RFA-OD-10-005中概述的五个主题中的两个：“应用基因组学和其他高通量技术”和“将基础科学发现转化为新的更好的治疗方法”，并准备立即实施。脑肿瘤是一个复杂的微观世界，肿瘤细胞和非肿瘤细胞之间的双向相互作用驱动着肿瘤的发生和发展。我们对肿瘤细胞和肿瘤周围之间复杂的相互作用的了解有限，这是充分认识脑肿瘤生物学的一个相当大的障碍，很可能是成功治疗脑肿瘤结果的一个重大障碍。我们实验室使用基因工程小鼠进行的研究表明，肿瘤微环境（基质）在决定胶质瘤形成和生长的时间和地点方面发挥了特殊作用。特别是，我们已经证明这些肿瘤中的非肿瘤免疫系统样细胞（小胶质细胞）精心制作了对胶质瘤形成和维持至关重要的特定生长促进因子。在本提案中，我们召集了一个具有跨学科科学专业知识和先前参与大规模合作研究计划的研究人员团队，并将他们的工作重点放在鉴定肿瘤微环境中存在的小胶质细胞产生的靶向分子以进行临床前评估。具体来说，我们计划扩展我们与华盛顿大学医学院基因组中心正在进行的研究，使用下一代测序和生物信息学管道来发现小鼠和人类胶质瘤相关小胶质细胞共享的新标记和信号，并代表基质定向治疗的潜在靶点（1期）。鉴定的基因将用于开发肿瘤相关小胶质细胞亚分类的免疫试剂（2a期），以及发现新的胶质瘤促进因子（“胶质瘤原”）（2b期）。在2a期开发的单克隆抗体接下来将用于将小胶质细胞亚群分类为功能不同的群体，并确定特定小胶质细胞群体与临床肿瘤行为的相关性（3a期）。此外，通过慢病毒操作在Nf1基因工程小鼠（GEM）菌株（2b期）中验证的先导小胶质细胞胶质瘤原将通过使用基质共培养系统的高通量技术进行评估，以确定基质导向的药物抑制剂，用于未来的临床前治疗研究（3b期）。

DESCRIPTION (provided by applicant): In summary, this proposal focuses on two of the five themes outlined in the RFA-OD-10-005: "Applying Genomics and Other High Throughput Technologies" and "Translating Basic Science Discoveries into New and Better Treatments" and is ready for immediate implementation. Brain tumors are complex microcosms in which bi-directional interactions between neoplastic cells and non-neoplastic cells drive tumorigenesis and tumor progression. Our limited knowledge of the complex interplay between tumor cells and the tumor surround is a considerable obstacle to fully appreciating brain tumor biology, and most likely represents a significant barrier to successful brain tumor treatment outcomes. Studies from our laboratories using genetically-engineered mice have demonstrated the obligate role of the tumor microenvironment (stroma) in dictating when and where gliomas form and grow. In particular, we have shown that resident non-neoplastic immune system-like cells (microglia) in these tumors elaborate specific growth-promoting factors critical for glioma formation and maintenance. In this proposal, we have assembled a team of investigators with expertise in cross-disciplinary science and prior involvement in large-scale cooperative research initiatives, and focused their efforts on identifying targetable molecules made by microglia present in the tumor microenvironment for preclinical evaluation. Specifically, we plan to extend our ongoing studies with The Genome Center at Washington University School of Medicine to use next-generation sequencing and bioinformatics pipelines to discover new markers and signals shared by mouse and human glioma-associated microglia, and which represent potential targets for stromal-directed therapy (Phase 1). Identified genes will be used to develop immunologic reagents for the subclassification of tumor-associated microglia (Phase 2a) as well as to discover novel glioma-promoting factors ("gliomagens") (Phase 2b). Monoclonal antibodies developed in Phase 2a will next be employed to classify microglia subsets into functionally distinct populations and to determine the relevance of specific microglia populations to clinical tumor behavior (Phase 3a). In addition, lead microglia gliomagens validated by lentiviral manipulation in Nf1 genetically-engineered mouse (GEM) strains (Phase 2b) will be evaluated by high-throughput technology using a stromal co-culture system to identify stromal-directed pharmacologic inhibitors for future preclinical therapeutic studies (Phase 3b). PUBLIC HEALTH RELEVANCE: Brain tumors (gliomas) are the leading cause of cancer-related death in children with few successful treatments available. This proposal applies genomics and novel high throughput technologies to translate basic science discoveries into new and better treatments for children with low-grade glioma.