Integrative Approaches to Stroma-Directed Glioma Therapy

说明(申请人提供)：总之，本提案侧重于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推动了肿瘤的发生和发展。我们对肿瘤细胞和肿瘤周围复杂相互作用的有限了解是充分理解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的重要障碍。我们的实验室使用基因工程小鼠进行的研究表明，肿瘤微环境(间质)在决定胶质瘤形成和生长的时间和地点方面具有特定的作用。特别是，我们已经证明了这些肿瘤中的常驻非肿瘤免疫系统样细胞(小胶质细胞)阐述了对胶质瘤的形成和维持至关重要的特定的生长促进因子。在这个提案中，我们组建了一个具有跨学科科学专业知识并参与过大规模合作研究计划的研究团队，并将他们的努力集中在识别存在于肿瘤微环境中的小胶质细胞制造的靶向分子上，以进行临床前评估。具体地说，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新的标记和信号，这些标记和信号与小鼠和人类胶质瘤相关的小胶质细胞共享，并代表着间质导向治疗的潜在靶点(第一阶段)。已确定的基因将被用于开发免疫试剂，用于肿瘤相关小胶质细胞的亚类(2a期)以及发现新的胶质瘤促进因子(“胶质瘤”)(2b期)。在2a期开发的单抗接下来将用于将小胶质细胞亚群分类为功能不同的群体，并确定特定的小胶质细胞群体与临床肿瘤行为的相关性(3a期)。此外，在NF1基因工程小鼠(2b期)品系中通过慢病毒操作验证的铅小胶质细胞胶质瘤细胞(2b期)将通过高通量技术进行评估，该技术使用基质共培养系统来识别基质导向的药理抑制剂，用于未来的临床前治疗研究(3b期)。 公共卫生相关性：脑瘤(胶质瘤)是儿童癌症相关死亡的主要原因，几乎没有成功的治疗方法。这项建议应用基因组学和新的高通量技术，将基础科学发现转化为治疗低级别胶质瘤儿童的新的、更好的治疗方法。

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.