Dissecting the role of mitochondrial glutathione homeostasis in cancer

项目摘要 所有真核细胞，无论是正常的还是癌症的，都需要有感知营养水平变化的能力， 确保它们有效地用于生存和增长。营养感应机制使细胞能够迅速适应 环境扰动，这是癌细胞克服不同代谢的一个特别重要的特征 沿转移级联的应力。虽然已经描述了许多营养传感机制，但如何 在亚细胞室中感觉到代谢物仍然是一个主要的悬而未决的问题。这个问题特别是 与氧化还原活性分子有关，如NAD和谷胱甘肽，它们表现出显著的异质性 分布在亚细胞间，并已被证明在癌症转移中发挥关键作用。 最近在线粒体代谢物转运体去孤构化方面的突破提供了前所未有的 有机会以亚细胞的精度探索这些代谢物的动力学和传感机制。在一个 最近发表的一项研究表明，SLC25A39是线粒体谷胱甘肽的关键转运体。 主要的抗氧化剂分子与癌症进展和转移有关。值得注意的是，有证据表明 SLC25A39受线粒体谷胱甘肽的反馈调节，可能是有效 转移性定植，暗示它参与了癌症克服代谢应激的适应机制 转移。 这一建议寻求更深入地了解细胞器谷胱甘肽代谢在癌症中的意义。 该提案的目的1试图了解线粒体谷胱甘肽稳态在肿瘤中的作用。 并破译其调控机制。这项提案的目标2旨在 开发新的遗传编码、单细胞RNAseq兼容的报告程序以分析细胞间 肿瘤中线粒体谷胱甘肽的异质性。结合生化分析，不偏不倚 CRISPR屏幕和新的动物模型，这项提案旨在描绘一幅多层次的动态图景， 线粒体谷胱甘肽稳态在肿瘤中的调节机制及其功能贡献 进展和转移。建议的研究完成后，将加深我们对 癌症代谢重排中的代谢物池划分及新疗法的启示 针对转移的策略。

Project Summary All eukaryotic cells, whether normal or cancerous, require the ability to sense changes in nutrients levels, ensuring their efficient use for survival and growth. Nutrient sensing mechanisms enable cells to rapidly adapt to environmental perturbation, a feature particularly essential for cancer cells to overcome diverse metabolic stresses along the metastatic cascade. Although many nutrient sensing mechanisms have been described, how metabolites are sensed in subcellular compartments remains a major open question. This question is particularly relevant for redox-active molecules such as NAD and glutathione, which display remarkably heterogenous distribution across subcellular compartments and have been shown to play key roles in cancer metastasis. Recent breakthroughs in deorphanizing mitochondrial metabolite transporters provided unprecedented opportunity to probe the dynamics and sensing mechanism of these metabolites at subcellular precision. In a recently published study, SLC25A39 has been identified as a key transporter for mitochondrial glutathione, a major antioxidant molecule implicated in cancer progression and metastasis. Remarkably, evidence suggests that SLC25A39 undergoes feedback regulation by mitochondrial glutathione and may be required for efficient metastatic colonization, implicating it in an adaptive mechanism for cancers to overcome metabolic stress during metastasis. This proposal seeks a deeper understanding of the implication of organellar glutathione metabolism in cancer. The Aim 1 of this proposal seeks to understand the role of mitochondrial glutathione homeostasis in tumor progression and metastasis and decipher the mechanism of its regulation. The Aim 2 of this proposal seeks to develop novel genetically encoded, single-cell RNAseq-compatible reporters for profiling intercellular heterogeneity in mitochondrial glutathione in tumors. Using a combination of biochemical analysis, unbiased CRISPR screens and novel animal models, this proposal aims to paint a multilayered picture of the dynamics, regulatory mechanisms and functional contribution of mitochondrial glutathione homeostasis in tumor progression and metastasis. Completion of the proposed studies will deepen our understanding on the role of compartmentalized metabolite pools in metabolic rewiring of cancers and shed light on novel therapeutic strategies to target metastasis.