The role of lysosomal deregulation in cancer progression

溶酶体失调在癌症进展中的作用

• 批准号: 10217504
• 负责人: Arash Latifkar
• 金额: $ 9.31万
• 依托单位: WHITEHEAD INSTITUTE FOR BIOMEDICAL RES
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-08-18 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10217504
• 关键词: Aggressive behavior; Animal Model; Behavior; Binding Proteins; Breast Cancer Cell; Cancer Model; Cathepsins; Cell Survival; Cells; Cessation of life; Clinical; DNA; Deacetylase; Disease; Disseminated Malignant Neoplasm; Down-Regulation; Environment; Excision; Fellowship; Foundations; Future; Generations; Goals; Grant; Half-Life; Health; Hydrolase; Impairment; Lysine; Lysosomes; Malignant - descriptor; Malignant Neoplasms; Mediating; Mediator of activation protein; Messenger RNA; Metastatic breast cancer; Metastatic to; MicroRNAs; Modeling; Mus; Neoplasm Metastasis; Oncologist; Outcome; Pathway interactions; Phase; Phenotype; Positioning Attribute; Post-Translational Protein Processing; Postdoctoral Fellow; Process; Property; Proteins; Proton Pump; Pump; RNA; RNA Stability; RNA-Binding Proteins; Regulation; Research; Research Personnel; Research Project Grants; Research Proposals; Research Training; Resistance; Role; SIRT1 gene; Therapeutic; Therapeutic Agents; Therapeutic Intervention; Transcript; Up-Regulation; cancer cell; cancer therapy; cancer type; exosome; extracellular vesicles; in vivo; intercellular communication; metastatic process; novel; novel therapeutic intervention; tumor; tumor microenvironment; tumor progression; vacuolar H+-ATPase

Project Summary Aggressive metastatic breast cancer is responsible for the deaths of more than 40,000 people per year in the U.S., despite the best efforts of researchers and clinical oncologists. I have recently discovered a potentially important mechanism regarding the generation of a unique secretome by breast cancer cells, which makes an essential contribution to their invasiveness and metastatic capability. Specifically, I have delineated an intriguing connection between the down-regulation of Sirtuin 1 (Sirt1), a NAD-dependent deacetylase, in aggressive breast cancer cells and the corresponding reduction in the expression of a major subunit of the vacuolar ATPase (V-ATPase), which results in the impairment of their lysosomes and consequently, dramatic changes in their secretome. These changes include a significant increase in the number of exosomes generated by breast cancer cells, and an enrichment in their ubiquitylated protein cargo. Exosomes are small (extracellular) vesicles, ~30-150 nm in size, that contain a wide range of cargo including proteins, RNA transcripts, microRNA, and even DNA. They function as mediators of intercellular communication and have been implicated in a number of aspects of cancer progression, including the promotion of chemo-resistance and the formation of a pre-metastatic niche. Because exosomes are also attractive vehicles for the delivery of therapeutic agents, studies aimed at determining how exosomes are formed and released, as well as characterizing their functional properties, are being extensively pursued. Thus, these findings now highlight how aggressive breast cancer cells generate exosomes containing unique cargo, which contribute to the metastatic capability of breast cancer cells. I further discovered that the down-regulation of Sirt1 in breast cancer cells results in a significant increase in the secretion of soluble hydrolases, in particular, cathepsins. Collectively, these components making up the secretome of aggressive breast cancer cells give rise to a marked enhancement in migratory and invasive activity. In the F99 phase of this application, these discoveries will be extended by determining the underlying mechanisms by which the down-regulation of Sirt1 in aggressive cancer cells leads to a reduced expression of the V-ATPase (Aim 1). A particular emphasis will be to identify the Sirt1 substrate that is directly responsible for regulating the stability of the RNA transcript encoding one of the major subunits of the V-ATPase. In the K00 phase of the proposal (Aim 2), a research/training environment will be sought to develop animal models that will further establish the functional connection between Sirt1, the v-ATPase and lysosomal function, and demonstrate how this contributes to the metastatic state. The ultimate goal of my studies will be to highlight strategies that alter these connections in a manner that leads to new anti-cancer therapies.

项目摘要 侵袭性转移性乳腺癌每年造成40，000多人死亡， 美国，尽管研究人员和临床肿瘤学家尽了最大的努力。我最近发现了一个潜在的 关于乳腺癌细胞产生独特分泌组的重要机制， 对它们的侵袭性和转移能力有重要贡献。具体来说，我描绘了一个 Sirtuin 1（Sirt 1），一种NAD依赖性去乙酰化酶， 侵袭性乳腺癌细胞和相应的减少表达的主要亚基的 空泡ATP酶（V-ATP酶），导致其溶酶体受损， 它们的分泌组发生变化。这些变化包括外泌体数量的显著增加 由乳腺癌细胞产生，并富集其泛素化蛋白质货物。外泌体很小 （细胞外）囊泡，大小约为30-150 nm，含有广泛的货物，包括蛋白质、RNA 转录物、微RNA甚至DNA。它们作为细胞间通讯的介质发挥作用， 与癌症进展的许多方面有关，包括促进化疗耐药性 和转移前小生境的形成。因为外泌体也是有吸引力的载体， 治疗剂，旨在确定外泌体如何形成和释放的研究，以及 表征其功能特性的方法正在被广泛追求。因此，这些发现现在强调 侵袭性乳腺癌细胞如何产生含有独特货物的外泌体， 乳腺癌细胞的转移能力。我进一步发现，乳腺癌中Sirt 1的下调 癌细胞导致可溶性水解酶，特别是组织蛋白酶的分泌显著增加。 总的来说，这些组成侵袭性乳腺癌细胞分泌组的成分引起了乳腺癌细胞的凋亡。 迁移和入侵活动显著增强。在本申请的F99阶段，这些发现 将通过确定Sirt 1下调的潜在机制来扩展， 侵袭性癌细胞导致V-ATP酶表达减少（Aim 1）。一个特别的重点将 鉴定直接负责调节RNA转录物稳定性的Sirt 1底物 编码V-ATP酶的主要亚基之一。在提案的K 00阶段（目标2）， 将寻求研究/培训环境来开发动物模型，以进一步建立功能性 Sirt 1，v-ATPase和溶酶体功能之间的联系，并证明这如何有助于 转移状态我研究的最终目标将是强调改变这些联系的策略， 一种能带来新的抗癌疗法的方式。