Biological Comparisons Among Three Derivative Models of Glioma Patient Cancers Under Microenvironmental Stress

项目摘要/摘要： 目前潜在治疗药物的临床前测试方法在很大程度上是平淡无奇的。 就其产生临床影响的能力而言。胶质母细胞瘤(GBM)尤其如此，其预后 在过去的75年里只增加了2-3个月，5年存活率不到4%。许多有希望的人 临床前研究在临床测试中未能达到预期。此问题可能是由于：a) 临床前模型系统的局限性和b)缺乏可靠的生物标记物来正确选择患者。至 为了解决这些问题，研究人员越来越多地利用患者衍生的癌症模型(PDMC) 具有全面的分子图谱。然而，模型组成、生长条件和 其他微环境因素限制了这些模型的可靠性，并阻碍了解释。我们相信，一个 在3种患者来源的肿瘤模型(异种)上仔细研究肿瘤微环境应激源 GBM，即异种移植(PDX)、球形培养(神经球)和植入3D的人生物基质 微肿瘤，将提供对受模型和TME影响的肿瘤生物学的关键方面的洞察。 这些TME扰动前后的模型也将在基因组学上得到全面的描述， 转录和动态学(通过肽底物微阵列评估全球激酶活性)水平 生成相似性距离度量。我们假设将TME应激源应用于PDMC的意志 提高各自模型的分子和生物学保真度，使之与原始肿瘤或亲本模型的保真度相比 可以使用先进的计算数据建模系统在硅胶中进行可视化和测试的磷灰石 (GeneTerrain)。我们从NIH以前资助的项目中产生的初步数据表明，我们现有的Xen oline 概述在癌症基因组图谱中发现的所有四种GBM分子亚型，同时复制 将GBM原位移植到免疫低下的小鼠体内时的关键特征。重要的是，我们可以发展 一种新的三维(3D)培养系统中的分解的异种肿瘤，该系统整合了许多 肿瘤微环境产生适合更高通量药物测试的3D微肿瘤。此外，我们 有初步证据表明TME处理BTICs或3D微肿瘤(例如缺氧或营养不良) 剥夺)在体外和体内促进更具侵袭性的肿瘤表型，并伴随着变化 在动态学签名中。因此，我们将：1)从现有的磷烯烃生成3个PDMC模型以及 Novo GBM患者肿瘤患者进行全面的基因组测试以计算 具有相应生物学评估的模型，包括生长、化学辐射敏感性和茎 标记物；;2)对衍生的PDMC(神经球和3D微肿瘤)进行TME扰动测试，以及 确定对肿瘤生物学和相似距离的影响；;3)开发和验证GeneTerrain模型 关于TME和治疗敏感性(辐射和替莫唑胺)的各种PDMC。

PROJECT SUMMARY/ABSTRACT: Current methods of preclinical testing of potential therapeutics have been, for the most part, underwhelming in terms of their ability to yield a clinical impact. This is particularly true for glioblastoma (GBM) where prognosis has increased only by 2-3 months over the last 75 years with a 5-year survival of less than 4%. Many promising preclinical studies have failed to live up to expectations when tested clinically. This problem is likely due to: a) limitations of the preclinical model system and b) lack of reliable biomarkers for proper patient selection. To address these issues, investigators are increasingly utilizing patient-­derived models of cancer (PDMC) coupled with comprehensive molecular profiling. However, differences in model composition, growth conditions, and other microenvironmental factors limit reliability of these models and hamper interpretation. We believe that a careful investigation of tumor microenvironmental (TME) stressors on 3 patient­derived models (xenolines) of GBM, namely xenografts (PDX), spheroid cultures (neurospheres), and human biomatrix embedded 3D microtumors, will provide insight into critical aspects of tumor biology that are influenced by model and TME. These models pre­ and post­ TME perturbagen will also be comprehensively profiled at the genomic, transcriptomic, and kinomic (global kinase activity assessment through a peptide substrate microarray) level to generate a similarity distance metric. We hypothesize that application of TME stressors to the PDMC’s will improve both molecular and biological fidelity of the respective models to that of the original tumor or parent xenoline that can be visualized and in silico tested using an advanced computational data modeling system (GeneTerrain). Our preliminary data generated from prior NIH funded projects indicate that our existing xenolines recapitulate all four molecular subtypes of GBM identified in The Cancer Genome Atlas while reproducing the key hallmarks of GBM when implanted orthotopically in immunocompromised mice. Importantly, we can grow disaggregated xenoline tumors in a novel three-dimensional (3D) culture system incorporating many cells of the tumor microenvironment to produce 3D microtumors suitable for higher throughput drug testing. Moreover, we have preliminary evidence that TME manipulation of BTICs or 3D microtumors (e.g., hypoxia or nutrient deprivation) promotes a more aggressive tumor phenotype in vitro and in vivo that is accompanied by changes in kinomic signatures. Therefore, we will: 1) Generate 3 PDMC models from existing xenolines as well as de novo GBM patient tumors with comprehensive omic testing to calculate similarity distance metrics among the models with corresponding biological assessments including growth, chemoradiation sensitivity, and stemness markers;; 2) Perform TME perturbagen testing of the derivative PDMCs (neurospheres and 3D microtumors) and determine impact on tumor biology and similarity distance metric;; 3) Develop and validate GeneTerrain models of the various PDMCs with respect to TME and therapeutic sensitivity (radiation and temozolomide).