Metabolic imaging comparisons of patient-derived models of renal cell carcinoma

今年，大约62,000名美国人将被诊断为肾癌，超过14,000人将被诊断为肾癌 死于这种疾病。十分之九的肾癌是肾细胞癌(RCC)。为了降低死亡率， 对于肾癌，从诊断到预后再到治疗，所有阶段都需要改进。为回应这一事件 资助机会“癌症患者衍生模型中的生物学比较(U01)”，我们将比较四个 患者衍生的肾癌模型的类型来调查每种模型作为临床前模型的相对真实性。 第一种将是患者来源的异种移植物(PDX)，被广泛认为是最具代表性的 人类病理生理学。这些都是先前从一系列病理和临床证实的。 RCC分期。PDX将作为比较三种PDX派生模型的“黄金标准”， 包括组织切片培养(TSC)、原代细胞培养和由细胞培养产生的异种移植。 我们关注的生物比较是新陈代谢。新陈代谢失调，其中之一 是癌症的特征，与肾癌的发生和发展密切相关。多效性变化 包括氧感知、能量感知和营养感知的失调。尤其是高频 VHL和FBP1基因的突变导致了“Warburg效应”(糖酵解的升高 氧的存在)，在透明细胞RCC，RCC的主要亚型，导致增加产量和 乳酸的排泄。比较四种患者衍生的肾癌模型的新陈代谢将捕捉到 遗传、转录、环境和其他影响的功能后果提供了 每个模型系统的表型的综合图片。我们将使用超极化(HP)13C磁体 磁共振(MR)，一种非常敏感的分子成像技术，用于监测动态通路特异性 患者来源的肾癌模型的代谢和生理过程，产生生物学和临床上的结果 相关数据。 目标1将通过HP MR成像和稳态来识别8个RCC PDX的代谢特征 代谢组学研究。代谢数据将与遗传型、转录型和免疫型相关。 用于确定每个PDX的表型的特征。在目标2中，将准备薄的精密切割的组织切片 从8个PDX中的每一个中提取并放置在与核磁共振兼容的3D组织培养生物反应器中。新陈代谢 TSCs的表型将通过HP MR和稳态研究来确定，并与 原始的PDX，以及遗传、转录和免疫组织学特征。类似的研究将会是 在目标3中使用来源于PDX的原代细胞培养，在目标4中使用由以下物质产生的异种移植 PDX来源的细胞培养物在小鼠体内的移植。在目标5中，将对四种类型的模型进行最终测试 临床相关谷氨酰胺酶抑制剂CB-839的代谢反应比较 进入RCC的临床试验。

This year, ~62,000 Americans will be diagnosed with kidney cancer and more than 14,000 individuals will die from this disease. Nine of ten kidney cancers are renal cell carcinoma (RCC). To reduce mortality from RCC, improvements are needed at all stages, from diagnosis to prognosis to therapy. In response to the funding opportunity “Biological Comparisons in Patient-derived Models of Cancer (U01)”, we will compare four types of patient-derived models of RCC to investigate the relative authenticity of each as a preclinical model. The first will be patient-derived xenografts (PDXs), widely perceived as the most representative models of human pathophysiology. These have been previously established from a range of pathologic and clinical stages of RCC. The PDXs will serve as the “gold standard” to which to compare three PDX-derived models, including tissue slice cultures (TSCs), primary cell cultures, and xenografts generated from cell cultures. The biological comparison on which we focus is metabolism. Dysregulated metabolism, one of the hallmarks of cancer, is strongly implicated in the development and progression of RCC. Pleiotropic changes include dysregulation of oxygen sensing, energy sensing and nutrient sensing. In particular, high frequency mutations in VHL and FBP1 genes contribute to exhibition of the “Warburg effect” (an elevation of glycolysis in the presence of oxygen) in clear cell RCC, the major subtype of RCC, leading to increased production and excretion of lactate. Comparing metabolism among the four patient-derived models of RCC will capture the functional consequences of genetic, transcriptomic, environmental and other influences to provide a comprehensive picture of the phenotype of each model system. We will use hyperpolarized (HP) 13C magnetic resonance (MR), a remarkably sensitive molecular imaging technique, to surveil dynamic pathway-specific metabolic and physiologic processes in the patient-derived RCC models, yielding biologically and clinically relevant data. Aim 1 will identify the metabolic signature of each of 8 RCC PDXs by HP MR imaging and steady state metabolomic profiling. The metabolic data will be associated with genotypic, transcriptomic and immunotypic features to establish the phenotype of each PDX. In Aim 2, thin precision-cut tissue slices will be prepared from each of the 8 PDXs and placed in a NMR-compatible, 3D tissue culture bioreactor. The metabolic phenotype of the TSCs will be determined by HP MR and steady state studies and compared to that of the original PDXs, along with genetic, transcriptomic and immunohistologic features. Similar studies will be performed in Aim 3 with primary cell cultures derived from PDXs, and in Aim 4 with xenografts generated by the implantation in mice of PDX-derived cell cultures. In Aim 5, the final test of the four types of models will be a comparison of metabolic responses to the clinically relevant glutaminase inhibitor CB-839, which is currently entering clinical trials in RCC.