Microenvironmental Nutrient Availability and Immunomodulation in Lung Cancer Cel

.项目总结（见说明）; 我们提出了一个系统的生化研究肺癌（LC）细胞营养代谢的重要性，LC的发展，生存和肿瘤微环境（TME）的进展。主要方法是利用稳定同位素（13 C和15 N）营养示踪剂结合稳定同位素分辨代谢组学（SIRM）和代谢组学编辑的转录组学分析（Meta）来识别LC细胞中的关键代谢事件，这些事件决定了LC细胞对主要TME因素（如缺氧和营养缺乏）的行为和脆弱性。这个以细胞为基础的项目的一个中心焦点是了解营养物质的可用性和缺氧在调节LC细胞代谢中的相互作用，以及这可能如何影响其生长，存活和进展的能力。这一关注部分是由我们最近从人类患者切除的成对癌性和良性肺组织的基因阵列数据中发现的关于参与谷氨酰胺和葡萄糖代谢的关键酶（例如，乙酰氨基酶、乙酰氨基酶、透明质酸合酶2）的失调的结果所激发的。这也是由于我们最近发现肺肿瘤异种移植物中的胰蛋白酶抑制和天然免疫激活剂B-葡聚糖激活小鼠巨噬细胞中的Gin代谢。我们将通过以下具体目标实现这一目标：SAI：肺细胞的SIRM谱，用于重建参与葡萄糖和谷氨酰胺利用的生化途径。这些营养物质的生化途径相关的能量，anastomosis，免疫调节传感器，和细胞迁移相关的细胞外基质因子将被映射在LC细胞窝藏主要驱动基因异常，正常上皮细胞，和相关的基质细胞进行比较。SA 2：探讨营养物质有效性与缺氧的相互作用。我们将重点关注两个关键问题：1）杜松子酒如何缓解LC细胞对葡萄糖的需求？; 2)Gin代谢对缺氧LC细胞至关重要吗？SAS：检查人肿瘤相关巨噬细胞中Arg代谢与Gln和Arg可用性的关系。将探测与肿瘤相关的人巨噬细胞中的免疫调节相关的Gin和Arg利用途径，以响应免疫激活剂（B-葡聚糖）和M2至Ml极化。从该项目中获得的知识将用于帮助解释从项目2和3中获得的数据。

. PROJECT SUMMARY (See instructions); We propose a systems biochemical study of lung cancer (LC) cell nutrient metabolism of importance to LC development, survival and progression in the tumor microenvironment (TME). The primary approach is to utilize stable isotopic (13C and 15N) nutrient tracers in conjunction with stable isotope-resolved metabolomics (SIRM) and metabolomics-edited transcriptomic analysis (META) to discern key metabolic events in LC cells that govern their behavior and vulnerability in response to major TME factors such as hypoxia and nutrient deficiency. A central focus of this cell-based project is to understand the interaction of nutrient availability and hypoxia in modulating LC cell metabolism and how this may affect its ability to grow, survive and progress. This focus is in part motivated by our recent finding from the gene array data of paired cancerous and benign lung tissues resected from human patients regarding the dysregulations of key enzymes (e.g. arginase, glutaminase, hyaluronan synthase 2) involved in metabolism of glutamine and glucose. It is also driven by our recent discovery of arginase suppression in lung tumor xenograft and activation of Gin metabolism in murine macrophages by a natural immune activator, B-glucan. We will accomplish this with the following specific aims: SAI: SIRM profiling of lung cells for reconstructing biochemical pathways involved in utilization of glucose and glutamine. Biochemical pathways of these nutrients relevant to energy, anabolism, immunomodulatory sensors, and cell migration-related extracellular matrix factors will be mapped in LC cells harboring major driver gene anomalies, normal epithelial cells, and relevant stromal cells for comparison. SA2: Probe interactions of nutrient availability in combination with hypoxia. We will focus on 2 key questions: 1) How does Gin alleviate glucose demand by LC cells?; 2) Is Gin metabolism crucial to hypoxic LC cells?. SAS: Examine Arg metabolism in human tumor-associated macrophages as a function of Gin and Arg availability. Gin and Arg utilization pathways of relevance to immunomodulation in tumor-associated human macrophages will be probed in response to immune activator (B-glucans) and M2 to Ml polarization. The knowledge gained from this project will be used to help interpret data obtained from Projects 2 and 3.