Microenvironmental Nutrient Availability and Immunomodulation in Lung Cancer Cel

。项目总结(见说明)； 我们建议对肺癌(LC)细胞的营养代谢进行系统的生化研究，该研究对肺癌的发生、生存和肿瘤微环境(TME)的进展具有重要意义。主要的方法是利用稳定同位素(13C和15N)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分析(META)来识别LC细胞中的关键代谢事件，这些代谢事件决定了LC细胞在应对主要TME因素(如缺氧和营养缺乏)时的行为和脆弱性。这个以细胞为基础的项目的中心焦点是了解营养可获得性和低氧在调节LC细胞代谢过程中的相互作用，以及这可能如何影响其生长、生存和发展的能力。这一关注的部分原因是我们最近从从人类患者切除的癌和良性肺组织的基因阵列数据中发现了参与谷氨酰胺和葡萄糖代谢的关键酶(如精氨酸酶、谷氨酰胺酶、透明质酸合成酶2)的失调。这也是因为我们最近发现在肺癌异种移植中抑制精氨酸酶，并通过天然免疫激活剂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来实现这一点：SAI：SIRM分析肺细胞，以重建与葡萄糖和谷氨酰胺利用有关的生化途径。这些营养物质与能量、合成代谢、免疫调节感受器和细胞迁移相关的细胞外基质因子相关的生化途径将被定位在存在主要驱动基因异常的LC细胞、正常上皮细胞和相关基质细胞中进行比较。SA2：探索与低氧相结合的养分有效性的相互作用。我们将集中讨论两个关键问题：1)Gin如何减轻LC细胞对葡萄糖的需求；2)Gin代谢对低氧LC细胞是否至关重要？SAS：检测人体肿瘤相关巨噬细胞的Arg代谢与Gin和Arg可用性的关系。在免疫激活剂(B-葡聚糖)和M2至M1极化的反应中，将探索与肿瘤相关的人巨噬细胞的免疫调节相关的GIN和Arg的利用途径。从这个项目中获得的知识将被用来帮助解释从项目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.