Pathway Analysis in Mouse Model for Astrocytoma via Systems Biology Approach

高级别星形细胞瘤是最常见的脑部恶性肿瘤，可供选择的治疗方法有限，而且已被证实与一般较差的临床结果有关。因此，提高对疾病病因学的理解是开发创新诊断和治疗策略的一条很有希望的途径。目前的建议包括CAPR(Van Dykes博士临床前研究中心)、UNC(Ryan Millers博士实验室)和ISB(Leroy Hoods博士实验室)之间更广泛合作努力的初步试点阶段，以表征星形细胞缺乏症过程中潜在的分子指标和途径，并确定有希望的癌症转化和治疗反应的早期预后标记。为此，该项目利用了小鼠可诱导疾病启动的优势，扰乱和/或监测癌症发生过程的能力，以及无偏见系统生物学方法的广泛力量。随着疾病的进展，各种最先进的分子技术将监测组织和血清中的基因组、转录组、蛋白质组和代谢动力学。在可能的情况下，将利用跨物种数据分析来确定研究结果与人类疾病的相关性。在过去的一年里，CAPR的科学家们开发和优化了育种方法，以保持高级别星形细胞瘤和胶质母细胞瘤的一种可诱导遗传模型的群体。该模型以复杂的遗传结构为特征，将几个突变的等位基因混合在一起，使pRb/p105/p130、K-ras和PTEN/Akt通路单独或以不同的组合出现功能障碍。已经设计并验证了所需的一套分析方法，以快速识别具有所需等位基因组合的单个动物，以及多个基于组织学、分子生物学和活体成像的临床相关终点，以监测疾病进展阶段。例如，CAPR采用了一种升级的方法，允许通过MRI模式同时对多个动物进行成像，从而使整个实验队列的多个纵向成像会话成为可能。在另一项实验中，CAPR的临床前评估小组制定了一种原位异种颅内移植模型，该模型使用了人胶质瘤细胞系或从荷瘤动物中分离的原代GBM细胞。这些原位模型已被用于几个合作项目，旨在研究纳米技术化合物(例如，与NCL合作加载紫杉醇的纳米乳剂)和通过体外筛选策略确定的候选药物(例如，与Reilly博士合作的SchweinFursins)在脑组织中的生物利用度。

High grade astrocytomas represent the most common brain malignancies with limited therapeutic options available and proven association with generally poor clinical outcomes. Improved understanding of disease etiology thus represents a promising avenue for developing innovative diagnostic and therapeutic strategies. The current proposal comprises the initial pilot phase of a broader collaborative effort among CAPR (Dr. Van Dykes Preclinical Research Center), UNC (Dr. Ryan Millers lab) and ISB (Dr. Leroy Hoods lab) to characterize molecular indicators and pathways underlying the astrocytomagenesis processes and to identify promising early prognostic markers of cancerous transformation and response to treatment. To this end, the project leverages the advantages of inducible disease initiation in the mouse, the ability to perturb and/or monitor the carcinogenesis process, and the extensive power of unbiased systems biology approach. A variety of state-of-the-art molecular technologies will monitor genome, transcriptome, proteome, and metabolom dynamics in tissues and blood serum as the disease progresses. Where possible, a cross-species data analysis will be exploited to determine the relevance of findings to human disease. During the past year, CAPR scientists developed and optimized breeding approaches to maintain the colony of one-of-a-kind inducible genetic model for high-grade astrocytoma and glioblastoma. The model features a complex genetic setup that mingles several mutant alleles to render dysfunctional pRb/p105/p130, K-ras, and PTEN/Akt pathways alone or in different combinations. The required set of assays to rapidly identify individual animals with desired allelic combinations has been designed and validates, along with multiple histology-, molecular biology-, and in vivo imaging-based clinically relevant endpoints to monitor disease progression stage. To exemplify, an upgraded approach allowing simultaneous imaging of multiple animals by MRI modality has been adopted by CAPR making feasible multiple longitudinal imaging sessions of the entire experimental cohort. In another line of experiment, the Preclinical Evaluation group of CAPR elaborated an orthotopic intracranial xenograft models employing either human glioma cell lines or primary GBM cells isolated from tumor bearing animals. These orthotopic models have been utilized in several collaborative projects aimed at bioavailability studies in brain tissue for nanotechnology compounds (e.g. paclitaxel loaded nanoemulsion, in collaboration with NCL) and drug candidates identified through in vitro screening strategies (e.g. Schweinfurthins, in collaboration with Dr. Reilly).