Project 2

项目2

• 批准号: 10676733
• 负责人: Gina Marie DeNicola
• 金额: $ 33.89万
• 依托单位: H. LEE MOFFITT CANCER CTR & RES INST
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-06-25 至 2026-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10676733
• 关键词: Alveolar; Cancer cell line; Catalysis; Cell Culture Techniques; Cell Death; Cell Proliferation; Cell Survival; Cells; Cessation of life; Compensation; Consumption; Culture Media; Cyst; Cystathionine; Cysteine; Cysteine Metabolism Pathway; Cystine; Cytoprotection; Dependence; Diet; Dipeptides; Energy Transfer; Environment; Enzyme Induction; Enzymes; Epithelial Cells; Exhibits; Genotype; Glutamates; Glutathione; Glycine; Goals; Immune; Impairment; KRASG12D; Liquid substance; LoxP-flanked allele; Lung; Lung Neoplasms; Malignant neoplasm of lung; Mediating; Metabolic; Metabolism; Mus; Non-Small-Cell Lung Carcinoma; Oxidation-Reduction; Play; Population; Primary carcinoma of the liver cells; Proliferating; Proteins; Resistance; Role; Serine; Serum; Source; Starvation; Sulfur; Testing; Therapeutic Intervention; Tissues; Treatment Efficacy; Work; cancer cell; deprivation; extracellular; in vivo; interstitial; lung cancer cell; metabolome; neoplastic cell; resistance mechanism; response; therapeutic target; transcription factor; tumor; tumor microenvironment; uptake

PROJECT 2 SUMMARY INVESTIGATING CYSTEINE ESSENTIALITY IN LUNG CANCER Cancer cells accumulate significant intracellular cysteine due to increased cystine uptake and loss of homeostatic control. Cysteine-derived molecules are crucial for cancer cell survival and proliferation as a consequence of their sulfur moiety, which facilitates diverse functions, including enzyme catalysis, energy transfer, and redox metabolism. Consistently, we have found that many lung cancer cell lines are highly sensitive to cystine starvation and lack the capacity for de novo cysteine synthesis, suggesting they are dependent on exogenous cystine as a source of cysteine. Importantly, the recently developed cyst(e)inase enzyme depletes extracellular cystine and cysteine efficiently, suggesting it may have therapeutic efficacy against lung cancer. However, intracellular metabolic circuits can modify the cellular response to starvation independent of transsulfuration capacity. Thus, understanding the compensatory mechanisms to cysteine starvation is crucial for efficient therapeutic targeting of cysteine in lung cancer. Importantly, our results suggest the glutathione synthesis machinery, an important target of the NRF2 transcription factor, has a glutathione-independent role to protect cells against cysteine starvation-induced ferroptosis. In addition, the tumor microenvironment has significant levels of alternative sources of cysteine, including glutathione, cystathionine and protein, that are lacking in cell culture and may promote survival in the absence of cystine. The central hypothesis of this proposal is that tissue of origin, metabolic responses to cystine starvation, and alteration in sulfur sources within the tumor microenvironment influence cystine essentiality in lung cancer, thereby leading to targetable metabolic vulnerabilities. We will test this hypothesis in the following specific aims: In Aim 1 we will evaluate the influence of tissue and cell of origin on cysteine metabolism and essentiality. In Aim 2 we will decipher metabolic circuits that dictate responsiveness to cysteine limitation. In Aim 3 we will evaluate cysteine metabolism in the tumor microenvironment. The ultimate goal, and the overall impact, of this project is to characterize key determinants of cysteine metabolism and dependence in lung cancer in order to define opportunities for therapeutic intervention.

项目 2 总结 研究半胱氨酸在肺癌中的重要性 由于胱氨酸的摄取增加和半胱氨酸的丢失，癌细胞在细胞内积累了大量的半胱氨酸。 稳态控制。半胱氨酸衍生分子对于癌细胞的存活和增殖至关重要 它们的硫部分的结果，促进多种功能，包括酶催化、能量 转移和氧化还原代谢。一致地，我们发现许多肺癌细胞系高度敏感 胱氨酸饥饿并且缺乏从头合成半胱氨酸的能力，表明它们依赖于 外源胱氨酸作为半胱氨酸的来源。重要的是，最近开发的半胱氨酸酶（cyst(e)inase）会消耗 细胞外胱氨酸和半胱氨酸有效，表明它可能对肺癌具有治疗功效。 然而，细胞内代谢回路可以改变细胞对饥饿的反应，独立于 转硫能力。因此，了解半胱氨酸饥饿的补偿机制至关重要 用于肺癌中半胱氨酸的有效治疗靶向。重要的是，我们的结果表明谷胱甘肽 合成机器是 NRF2 转录因子的重要靶标，具有不依赖于谷胱甘肽的作用 保护细胞免受半胱氨酸饥饿诱导的铁死亡。此外，肿瘤微环境还 大量的半胱氨酸替代来源，包括谷胱甘肽、胱硫醚和蛋白质， 缺乏细胞培养物，在缺乏胱氨酸的情况下可能会促进存活。本提案的中心假设 是组织的起源、对胱氨酸饥饿的代谢反应以及肿瘤内硫源的改变 微环境影响肺癌中胱氨酸的必需性，从而导致靶向代谢 漏洞。我们将在以下具体目标中检验这一假设： 在目标 1 中，我们将评估组织和细胞来源对半胱氨酸代谢和重要性的影响。 在目标 2 中，我们将破译决定对半胱氨酸限制反应的代谢回路。 在目标 3 中，我们将评估肿瘤微环境中的半胱氨酸代谢。 该项目的最终目标和总体影响是表征半胱氨酸的关键决定因素 肺癌的代谢和依赖性，以确定治疗干预的机会。