Defining metabolic adaptations within the PDAC "arid" tumor microenvironment

定义 PDAC“干旱”肿瘤微环境中的代谢适应

• 批准号: 10205608
• 负责人: CHI V. DANG
• 金额: $ 59.87万
• 依托单位: WISTAR INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-05-01 至 2026-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10205608
• 关键词: Affect; Amino Acids; Animal Model; Area; Back; Behavior; Biomass; Blood Vessels; Branched-Chain Amino Acids; CRISPR screen; Cancer Etiology; Cell Culture Techniques; Cell Proliferation; Cell division; Cells; Cellular Metabolic Process; Cessation of life; Citric Acid Cycle; Clinical; DNA Damage; Dependence; Diffusion; Environment; Generations; Genes; Glucose; Goals; Growth; Heterogeneity; Human; Hypoxia; Immune; In Vitro; Individual; Inter-tumoral heterogeneity; Label; Lipids; Malignant Neoplasms; Metabolic; Methods; Modeling; Molecular; Myeloid-derived suppressor cells; Nutrient; Oxidative Phosphorylation; Oxygen; Pancreatic Ductal Adenocarcinoma; Pathway interactions; Pharmaceutical Preparations; Phenotype; Production; Protocols documentation; Resources; Serum; Source; System; Testing; Therapeutic; Translations; Warburg Effect; aerobic glycolysis; base; cancer cell; cancer type; cell growth; chemotherapy; deprivation; experimental study; genome wide screen; in vivo; inhibitor/antagonist; insight; macromolecule; metabolic abnormality assessment; metabolic profile; metabolomics; neoplastic cell; nutrient deprivation; pancreatic cancer cells; pancreatic ductal adenocarcinoma cell; pancreatic ductal adenocarcinoma model; pancreatic neoplasm; pancreatic stellate cell; refractory cancer; response; standard of care; targeted treatment; therapy resistant; tumor; tumor microenvironment; whole genome

PROJECT SUMMARY The goal of our proposal is to identify therapeutic vulnerabilities associated with nutrient-poor conditions in tumors. Pancreatic ductal adenocarcinoma (PDAC) remains a major clinical challenge and is characterized by a dense stroma and paucity of blood vessels. The resulting hypoxia and scarcity of nutrients forces cancer cells to seek alternative sources of nutrients for growth. Paradoxically, cells residing in these nutrient- deprived microenvironments are among the most resistant to therapy as a result of poor drug diffusion and reduced cell proliferation (rendering DNA-damaging agents less effective). To target these cells, therefore, a better understanding of the adaptive mechanisms by which they cope with nutrient deprivation is urgently needed. In preliminary studies, we developed a culture protocol – limiting for oxygen, glucose, amino acids, and serum – to model the “arid conditions” of pancreatic tumors. Under such conditions, tumor cells slow down proliferation and metabolic activity. Using this system, we performed a whole genome CRISPR screen to identify genes required under arid conditions but dispensable under nutrient-replete (“fertile”) conditions. This revealed a strong dependency on the TCA cycle and oxidative phosphorylation for survival, but also suggested that genes involved in many biosynthetic activities (e.g. translation and cell division) are detrimental under arid conditions. Based on these results, we hypothesize that cells survive severe nutrient and oxygen deprivation by maximizing energy generation from limited fuel supplies while minimizing the use of such resources for macromolecule production. We further hypothesize that nutrient and oxygen deprivation alters the dynamic crosstalk between cancer cells and other cells comprising the tumor microenvironment (TME). Importantly, individual tumors may respond to these metabolic changes in different ways, contributing to inter-tumoral heterogeneity. Our proposal will explore these ideas to develop strategies that target therapy-resistant cancer cells residing in these nutrient-deprived niches. Aim 1. Characterize the heterogeneity in metabolic adaptations to arid conditions. Aim 2. Determine how arid conditions influence cellular crosstalk in the TME. Aim 3. Test the activity of metabolic inhibitors against cells in arid conditions in vitro and in vivo.

项目摘要 我们建议的目的是确定与营养贫困条件相关的治疗脆弱性 肿瘤。胰腺导管腺癌（PDAC）仍然是一个主要的临床挑战，其特征是 血管的致密基质和缺乏血管。产生的缺氧和营养素的稀缺性迫使癌症 细胞寻求替代养分生长的来源。矛盾的是，居住在这些养分中的细胞 - 由于药物扩散较差，因此被剥夺的​​微环境是对治疗最具耐药性的一种 并降低细胞增殖（使DNA损害剂的有效性降低）。因此，要靶向这些细胞 对他们应对营养剥夺的自适应机制有更好的理解是紧急的 需要。 在初步研究中，我们制定了一种培养方案 - 氧气，葡萄糖，氨基酸和系列的限制 - 建模胰腺肿瘤的“干旱条件”。在这种情况下，肿瘤细胞慢下来 增殖和代谢活性。使用此系统，我们执行了整个基因组CRISPR屏幕 确定在干旱条件下需要的基因，但在养分恢复（“肥沃”）条件下可分配。这 揭示了对TCA循环和氧化磷酸化的强烈依赖性，但也建议 参与许多生物合成活性（例如翻译和细胞分裂）的基因在干旱下是有害的 状况。基于这些结果，我们假设细胞在严重的营养和氧气中存活 通过从有限的燃料供应中最大化能源发电，同时最大程度地减少了此类资源的使用 大分子生产。我们进一步假设营养和氧气剥夺会改变动态 癌细胞与完成肿瘤微环境（TME）的其他细胞之间的串扰。重要的是， 单个肿瘤可能会以不同的方式对这些代谢变化做出反应，从而导致腹部间 异质性。我们的建议将探索这些想法，以制定针对抗治疗的策略 居住在这些养分缺乏的壁ches中的癌细胞。 目的1。表征对干旱条件的代谢适应中的异质性。 目标2。确定干旱条件如何影响TME中的细胞串扰。 目标3。在干旱条件下在体外和体内测试对细胞的代谢抑制剂的活性。