Metabolic regulation of exosome biogenesis as a determinant of cancer cell metastasis.

外泌体生物发生的代谢调节作为癌细胞转移的决定因素。

• 批准号: 10367331
• 负责人: MARC A ANTONYAK
• 金额: $ 41.74万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-02-01 至 2027-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10367331
• 关键词: 3-Dimensional; Acetylation; Affect; Automobile Driving; Biochemical; Biogenesis; Biological Models; Biology; Breast Cancer Cell; Breast Cancer Model; Breast cancer metastasis; Cancer Biology; Cathepsins; Cell surface; Cells; Cellular Assay; Characteristics; Chemicals; Cues; Databases; Deacetylase; Dependence; Development; Disease; Down-Regulation; Ensure; Exhibits; Generations; Glutamine; Goals; Immune response; Immunotherapy; Impairment; Laboratories; Lead; Light; Lung; Maintenance; Malignant Neoplasms; Mediating; Metabolic; Metabolism; Monitor; Multivesicular Body; Mus; Nature; Neoplasm Metastasis; Organoids; Play; Process; Production; Proteins; RNA; RNA-Binding Proteins; Reagent; Regulation; Resistance; Role; SIRT1 gene; Signal Transduction; Sirtuins; System; Testing; Therapeutic; Transcript; Work; Xenograft Model; addiction; aggressive breast cancer; base; cancer cell; design; exosome; expectation; extracellular vesicles; high resolution imaging; insight; malignant breast neoplasm; metastatic process; migration; mouse model; multidisciplinary; neoplastic cell; novel; three dimensional cell culture; trafficking; treatment strategy; tumor; tumor growth; tumor metabolism; tumor progression; vacuolar H+-ATPase

Abstract. The studies outlined in this proposal focus on the mechanisms by which aggressive breast cancer cells generate large numbers of exosomes with unique cargo, together with a total secretome that significantly enhances their potential for metastatic spread. They are based on exciting developments in the cancer biology field which show that exosomes, a major class of extracellular vesicles (EVs), play important roles in a number of aspects of cancer progression. These include the ability of exosomes to confer tumor cells with the capability to show resistance to chemotherapeutic reagents as well as to immune therapy, together with their roles in promoting metastatic spread. We recently discovered that the down- regulation of SIRT1 by aggressive breast cancer cells has an important influence on the numbers of exosomes that they generate, the nature of the exosome cargo, as well as the composition of their total secretome. This is due to the NAD+-dependent deacetylase/deacylase Sirtuin (SIRT1) playing a key role in maintaining normal lysosomal function through a novel mechanism that ensures the proper expression of a major subunit of the vacuolar ATPae (v-ATPase). We also have recently found that the formation and shedding of exosomes appear to be dependent on the elevations in glutamine metabolism characteristic of breast cancer cells (i.e. their ‘glutamine addiction’). These findings now raise important questions regarding how the dependence of aggressive breast cancer cells on glutamine metabolism influences and/or works together with the down-regulation of SIRT1 expression/activation to regulate lysosomal function and exosome biogenesis, thus producing a secretome that stimulates cancer cell invasiveness and helps drive the metastatic process. The different laboratories participating in this proposal will take advantage of their multi-disciplinary expertise in biochemical and chemical biology approaches in probing cancer cell metabolism and exosome biogenesis, high-resolution imaging, 3D spheroid culture and tumor organoids, and the use of mouse models, in probing three key aspects of the mechanisms driving breast cancer metastasis. These are: 1) Examining the relationship between SIRT1 down-regulation, elevated glutamine metabolism and the generation exosomes with unique cargo by aggressive breast cancer cells. 2) Understanding how SIRT1 down-regulation impacts vacuolar ATPase expression to generate a secretome capable of promoting cancer cell invasiveness. 3) Determining how SIRT1 expression/activity affects exosome production, cell invasiveness and metastatic spread in breast cancer models. The expectation is that these studies will lead to the identification of exciting new treatment strategies for the devastating effects of aggressive breats cancers, and ultimately, for other metastatic diseases.

抽象的。该提案中概述的研究重点是侵略性乳腺癌的机制 细胞产生大量带有独特货物的外泌体，以及一个总体秘密的外泌体 显着增强了它们的转移扩散潜力。他们基于令人兴奋的发展 癌症生物学领域表明外泌体是一类细胞外蔬菜（EV） 在癌症进展的许多方面的重要作用。这些包括外泌体的能力 会议肿瘤细胞具有表现出对化学治疗试剂的耐药性和免疫的能力 治疗，以及它们在促进转移性扩散中的作用。我们最近发现 - 通过侵略性乳腺癌细胞对SIRT1的调节对数量的数量有重要影响 它们产生的外泌体，外泌体货物的本质以及其总体的组成 秘密。这是由于NAD+依赖性脱乙酰基酶/脱乙酰酶Sirtuin（SIRT1）在 通过一种新的机制来维持正常的溶酶体功能，以确保适当的表达 真空ATPAE（V-ATPase）的主要亚基。我们最近还发现，形成和 外泌体的脱落似乎取决于谷氨酰胺代谢特征的高程 乳腺癌细胞（即“谷氨酰胺成瘾”）。这些发现现在提出了有关的重要问题 侵袭性乳腺癌细胞对谷氨酰胺代谢的依赖性如何影响和/或起作用 以及SIRT1表达/激活的下调以调节溶酶体功能和 外泌体生物发生，因此产生了刺激癌细胞侵袭性并有助于驱动的分泌组 转移过程。参与该建议的不同实验室将利用他们的优势 生化和化学生物学方法探索癌细胞的多学科专业知识 代谢和外泌体生物发生，高分辨率成像，3D球形培养和肿瘤类器官， 以及鼠标模型的使用，在探测驱动乳腺癌机制的三个关键方面 转移。这些是：1）检查SIRT1下调，升高谷氨酰胺之间的关系 侵略性乳腺癌细胞具有独特货物的代谢和产生外泌体。 2） 了解SIRT1下调如何影响真空ATPase表达以产生分泌组 能够促进癌细胞侵入性。 3）确定SIRT1表达/活动如何影响 乳腺癌模型中的外泌体产生，细胞侵袭性和转移性扩散。期望是 这些研究将导致确定令人震惊的灾难性效果的激动人心的新疗法 激进的呼吸癌，最终用于其他转移性疾病。