SIRT1 Regulates RNA Stability to Promote Breast Cancer

SIRT1 调节 RNA 稳定性促进乳腺癌发生

• 批准号: 10674902
• 负责人: Fangyu Wang
• 金额: $ 4.42万
• 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: 2022-08-01 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10674902
• 关键词: 3' Untranslated Regions; Acetylation; Aging; Area; Binding; Biogenesis; Breast Cancer Cell; Cancer Biology; Cancer Cell Growth; Catalytic Domain; Cell Aging; Cell Survival; Cell membrane; Cell physiology; Cell surface; Cells; Cessation of life; Deacetylase; Development; Disease; Disease Progression; Down-Regulation; Ectopic Expression; Exonuclease; Extracellular Space; Goals; Grant; Hydrolase; Impairment; Invaded; Laboratories; Learning; Longevity; Lysine; Lysosomes; Malignant Neoplasms; Mediating; Multivesicular Body; Neoplasm Metastasis; Pathway interactions; Patients; Persons; Phase; Play; Postdoctoral Fellow; Process; Production; Proteins; RNA; RNA Stability; RNA-Binding Proteins; Research; Research Personnel; Research Project Grants; Resistance; Risk Factors; Role; SIRT1 gene; Therapeutic; Time; Transcript; Tumor Promotion; Tumor Suppressor Proteins; Woman; Work; aged; aggressive breast cancer; breast cancer progression; cancer cell; cell age; cell growth; exosome; expectation; extracellular vesicles; graduate school; graduate student; insight; intercellular communication; interest; knock-down; malignant breast neoplasm; migration; new therapeutic target; novel; novel strategies; prevent; protein expression; recruit; senescence; skills; small hairpin RNA; trafficking; trait; transcriptome sequencing; tumor growth; tumor progression; vacuolar H+-ATPase

Project Summary Breast cancer is responsible for the deaths of more than 600,000 women each year. Thus, there is an over- riding need to better understand the mechanisms that promote breast cancer, as this information can potentially lead to new strategies to treat the disease. Recently, the Cerione lab discovered a mechanism through which highly aggressive forms of breast cancer cells produce a secretome containing factors that promote cancer cell survival and metastatic spread. Specifically, the researchers showed that the decreased expression of the NAD+- dependent deacetylase Sirtuin 1 (SIRT1) increases the formation and release of exosomes, a specific class of extracellular vesicles (EVs) that are derived from multi-vesicular bodies (MVBs) within the endocytic/lysosomal trafficking pathway. This effect was due to the increased acetylation of the RNA binding protein, IGF2BP2, that binds to the 3' untranslated region of the transcript encoding ATP6V1A, a major catalytic subunit of the vacuolar ATPase (v-ATPase) that maintains the proper pH and activity of lysosomes. Acetylated IGF2BP2 recruits an exonuclease, XRN2, which degrades the ATP6V1A transcript, resulting in impaired lysosomal activity. Thus, MVBs that would otherwise be targeted and degraded in lysosomes are instead directed to the cell surface where they fuse with the plasma membrane and release their contents, i.e., exosomes enriched in unique cargo and soluble hydrolases, into the extracellular space. These components of the secretome were shown to work together to strongly promote the invasiveness of breast cancer cells. Whether there were additional transcripts that were regulated similarly to the ATP6V1A transcripts by SIRT1 and IGF2BP2 were then determined. Based on a comprehensive RNA sequencing analysis performed, fourteen transcripts were identified; four of which encode proteins that potentially play important roles in breast cancer progression and/or exosome biogenesis. For the F99 phase of this application, I plan to establish that the stability of these transcripts is indeed regulated by SIRT1 and IGF2BP2, and to investigate how changes in their levels impact breast cancer progression (Aim 1). In addition to the tumor suppressor role played by SIRT1 in breast cancer, reductions in its expression have also been heavily implicated in aging, a major risk factor for developing cancer, as well as other diseases. In the K00 phase of this proposal, I would like to investigate the interplay between aging and cancer. It has been recently shown that the accumulation of aged/senescent cells results in the production of a secretome that can increase the growth of cancer cells. My plan as a Post-doctoral trainee will be to determine whether the EVs from aged/senescent cells contribute to this effect. Furthermore, I am also interested in exploring whether the EVs secreted from cancer cells enable cells to evade undergoing senescence (Aim 2). Ultimately, the goal of this study is to understand the relationship between aging and cancer to help identify potential therapeutic treatment for patients.

项目概要 乳腺癌每年导致超过 600,000 名女性死亡。因此，存在一个过度 骑行需要更好地了解促进乳腺癌的机制，因为这些信息可能会 导致治疗该疾病的新策略。最近，Cerione 实验室发现了一种机制 高度侵袭性的乳腺癌细胞产生含有促进癌细胞生长的因子的分泌蛋白组 生存和转移扩散。具体来说，研究人员表明 NAD+- 的表达减少 依赖的去乙酰化酶 Sirtuin 1 (SIRT1) 会增加外泌​​体的形成和释放，外泌体是一类特定的 细胞外囊泡 (EV) 源自内吞/溶酶体内的多囊泡体 (MVB) 贩运途径。这种效应是由于 RNA 结合蛋白 IGF2BP2 的乙酰化增加所致， 与编码 ATP6V1A 的转录本的 3' 非翻译区结合，ATP6V1A 是液泡的主要催化亚基 维持溶酶体适当 pH 值和活性的 ATP 酶（v-ATP 酶）。乙酰化 IGF2BP2 招募 核酸外切酶 XRN2 会降解 ATP6V1A 转录物，导致溶酶体活性受损。因此， 否则，MVB 将在溶酶体中被靶向和降解，而是被引导至细胞表面，其中 它们与质膜融合并释放其内容物，即富含独特货物的外泌体和 可溶性水解酶，进入细胞外空间。分泌蛋白组的这些成分被证明可以发挥作用 共同有力地促进乳腺癌细胞的侵袭。是否有额外的文字记录 然后确定了与 SIRT1 和 IGF2BP2 的 ATP6V1A 转录本类似的调节。基于 经过全面的 RNA 测序分析，鉴定出了 14 个转录本；其中四个 编码可能在乳腺癌进展和/或外泌体生物发生中发挥重要作用的蛋白质。 对于本申请的 F99 阶段，我计划确定这些转录本的稳定性确实受到调节 SIRT1 和 IGF2BP2，并研究其水平的变化如何影响乳腺癌进展（目标 1). SIRT1 除了在乳腺癌中发挥肿瘤抑制作用外，其表达的减少还导致 也与衰老密切相关，衰老是患癌症以及其他疾病的主要危险因素。在 在这个提案的K00阶段，我想研究衰老和癌症之间的相互作用。它一直 最近表明，衰老/衰老细胞的积累会导致分泌蛋白组的产生，该分泌蛋白组可以 增加癌细胞的生长。作为博士后实习生，我的计划是确定电动汽车是否 来自衰老/衰老细胞的细胞有助于这种效果。此外，我也有兴趣探索是否 癌细胞分泌的 EV 使细胞能够逃避衰老（目标 2）。最终的目标是 这项研究旨在了解衰老与癌症之间的关系，以帮助确定潜在的治疗方法 为患者提供治疗。