Research Testbed 2

摘要 脑瘤仍然是最恶性和最致命的癌症之一，成人胶质母细胞瘤(GBM)具有 中位生存期15个月，五年生存率不到10%(Stupp等人，2009年)，尽管做了手术， 化疗(替莫唑胺)和放射治疗。虽然这些方法中的每一种都提供了总体生存 几个月的好处，他们不提供更长期的生存好处或治愈，即使是组合。因此，新的 治疗方法是迫切需要的。要合理地开发治疗方法，关键是我们有更好的 基础和临床前研究的实验系统，忠实地概括了关键特征 同时将最先进的工程方法的全部力量应用于 建模和仿真。在未发表的工作中，我们发现睡美人(SB)转座酶系统 我们开发的可用于产生两种主要的GBM，原神经(PN)和间充质(MES)，在免疫- 使用已知的人胶质母细胞瘤致癌驱动因素的合格野生型小鼠，以及单个癌细胞 可通过多通道荧光成像在活体肿瘤中进行追踪。我们假设机械力 调节PN和MES亚型之间的关键靶向差异，以及免疫冷/热特征 PN和MES子类型代表了子类型之间的第二个关键目标差异。为了测试这些 假设，我们将测量活的GBM肿瘤中癌细胞和T细胞的细胞力和信号动力学 (目标1)并测量活体基底膜肿瘤的免疫细胞动力学(目标2)。特别是，我们将测量细胞牵引力 在活体肿瘤和脑组织中的力量，激酶和全球环境基金信号，以及细胞外基质结构。此外, 我们将跟踪抗肿瘤T细胞、前列腺癌小胶质细胞和骨髓来源的巨噬细胞，以及它们的 在PN和MES的SB小鼠模型中，相互之间和GBM细胞之间的相关性。这些实验将 用于细胞迁移模拟器(CMS)和布朗动力学肿瘤模拟器(BDTS)的参数化 创建特定于GBM子类型的预测模型。在大多数情况下，这些测量将是第一次 它们在活的GBM肿瘤中的种类，并将提供细胞和肿瘤规模的生物物理模型。通过这样做，RTB2将 正在为成像的发展提供最先进的实验-计算一体化试验台 技术部门的医疗模式。

Abstract Brain tumors remain among the most malignant and deadly cancers, with adult glioblastoma (GBM) having a median survival of 15 months and five-year survival of less than 10 percent (Stupp et al., 2009), despite surgery, chemotherapy (temozolomide), and radiation. While each of these approaches provides an overall survival benefit of a few months, they do not offer a longer-term survival benefit or cure, even in combination. Thus, new therapy approaches are sorely needed. To rationally develop therapies, it is critical that we have better experimental systems for both fundamental and preclinical investigations that faithfully recapitulate key features of the human disease while bringing the full power of state-of-the-art engineering approaches to bear including modeling and simulation. In unpublished work, we have found that the Sleeping Beauty (SB) transposase system we developed can be used to produce two major GBM, proneural (PN) and mesenchymal (MES), in immune- competent wild-type mice using known human oncogenic drivers of glioblastoma, and that individual cancer cells can be tracked in live tumors via multichannel fluorescence imaging. We hypothesize that mechanical forces mediate a key targetable difference between PN and MES subtypes, and that the immune cold/hot signatures of PN and MES subtypes represents a second key targetable difference between subtypes. To test these hypotheses, we will measure cellular force and signaling dynamics of cancer cells and T cells in live GBM tumors (Aim 1) and Measure immune cell dynamics in live GBM tumors (Aim 2). In particular we will measure cell traction forces, kinase and GEF signaling, and extracellular matrix architecture in live tumors and brain tissue. In addition, we will track antitumoral T cells, and protumoral microglia and bone marrow-derived macrophages, and their correlations with each other and GBM cells, in the SB mouse models of PN and MES. These experiments will be used to parameterize the Cell Migration Simulator (CMS) and Brownian Dynamics Tumor Simulator (BDTS) to create predictive models that are GBM subtype-specific. In most cases, these measurements will be the first of their kind in live GBM tumors, and will inform cell and tumor scale biophysical models. In so doing, RTB2 will be providing a state-of-the-art integrated experimental-computational testbed for development of imaging modalities in the TECH unit.