Targeting oncogenes for hepatocellular carcinoma

靶向肝细胞癌的癌基因

• 批准号: 9904586
• 负责人: DEVANAND SARKAR
• 金额: $ 40.21万
• 依托单位: VIRGINIA COMMONWEALTH UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-04-01 至 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9904586
• 关键词: Astrocytes; Automobile Driving; Binding; Biological Assay; Cancer Etiology; Cell Nucleus; Cessation of life; Clinical; Dendrimers; Development; Endoplasmic Reticulum; Genes; Hepatocyte; Human; Human Cell Line; Knock-out; Knowledge; Malignant Epithelial Cell; Malignant Neoplasms; Membrane; Membrane Proteins; Messenger RNA; Micrococcal Nuclease; Molecular; Molecular Analysis; Monitor; Mus; N-terminal; Oncogenes; Oncogenic; Pathogenesis; Pathway interactions; Patients; Play; Post-Transcriptional Regulation; Pre-Clinical Model; Preventive; Primary carcinoma of the liver cells; Proteins; Protocols documentation; RIPK1 gene; RNA; RNA Interference Therapy; RNA-Induced Silencing Complex; Role; Scaffolding Protein; Signaling Molecule; Site; Small Interfering RNA; System; TRAF2 gene; Therapeutic; Transgenic Organisms; Translating; Translational Regulation; Translations; Transmembrane Domain; Treatment Efficacy; Xenograft procedure; advanced disease; base; combinatorial; design; effective therapy; expectation; experimental study; in vivo; innovation; knock-down; mouse model; mutant; nanocomplexes; nanoparticle delivery; novel; nuclease; overexpression; pre-clinical; protein activation; recruit; secretory protein; targeted treatment; ubiquitin ligase

Summary Hepatocellular carcinoma (HCC), the fifth most common cancer and the second most common cause of cancer-related deaths worldwide, has no effective treatment for advanced disease. The present proposal focuses on two interacting proteins, AEG-1 and SND1, which function as bona fide oncogenes for HCC. AEG-1 and SND1 cooperate to increase RNA-induced silencing complex (RISC) activity where AEG-1 functions as a scaffold protein and SND1 functions as a nuclease. However, there is a gap of knowledge in our understanding of the mechanism(s) driving AEG-1 and SND1 cooperative oncogenic functions. In primary hepatocytes, AEG- 1 is predominantly localized in the nucleus, while in HCC cells AEG-1 is primarily localized in the endoplasmic reticulum (ER) membrane. Preliminary results indicate ER-anchoring is required for AEG-1 oncogenic function. ER-anchored AEG-1 binds specifically to secretory and membrane protein-encoding mRNAs to facilitate their translation. Analysis of SND1 RNA-interactome also identifies membrane protein-encoding mRNAs. Co- localization studies show that both AEG-1 and SND1 are located on ER membrane in HCC cells. Both AEG-1 and SND1 activate NF-κB. ER-anchored AEG-1 functions as a platform for upstream signaling molecules of NF-κB pathway and thus plays an essential role in NF-κB activation. The mechanism by which SND1 activates NF-κB is not known. We hypothesize that in transformed hepatocytes, AEG-1 translocates from the nucleus and anchors into the ER membrane where it recruits SND1 and both cooperate to promote HCC by modulating post-transcriptional regulation of mRNAs in RISC, translational regulation of membrane proteins and activation of NF-κB, AEG-1 and SND1 require each other for optimum functioning and might not exert oncogenic activity alone, and combinatorial inhibition of AEG-1 and SND1 might be an effective therapeutic strategy for HCC. Experiments are designed to interrogate these hypotheses using novel mouse models and targeted nanoplexes delivering siRNA for AEG-1 and SND1. This proposal will contribute to our long-term objectives of identifying key players regulating HCC pathogenesis and translating this knowledge into development of novel and effective targeted therapies. The immediate objective of the proposal is in-depth understanding of the molecular mechanisms by which AEG-1 and SND1 promote HCC and evaluate a combinatorial strategy of inhibiting AEG-1 and SND1 in a mouse model as a potential therapeutic. Thus the proposal has both mechanistic and therapeutic significance and innovation. Successful completion of the proposed studies will establish new targets for developing therapeutics and provide pre-clinical evidence for a targeted protocol.

概括 肝细胞癌 (HCC)，第五大常见癌症，也是第二大常见原因 全球范围内与癌症相关的死亡人数较多，目前尚无针对晚期疾病的有效治疗方法。目前的建议 重点关注两种相互作用的蛋白 AEG-1 和 SND1，它们是 HCC 的真正癌基因。 AEG-1 和 SND1 合作增加 RNA 诱导的沉默复合物 (RISC) 活性，其中 AEG-1 充当 支架蛋白和 SND1 充当核酸酶。然而，我们的理解存在知识差距 驱动 AEG-1 和 SND1 协同致癌功能的机制。在原代肝细胞中，AEG- 1 主要定位于细胞核，而在 HCC 细胞中 AEG-1 主要定位于内质 网状（ER）膜。初步结果表明 ER 锚定是 AEG-1 致癌功能所必需的。 内质网锚定的 AEG-1 与分泌蛋白和膜蛋白编码 mRNA 特异性结合，以促进其发挥作用 翻译。 SND1 RNA 相互作用组的分析还鉴定了膜蛋白编码 mRNA。共同 定位研究表明，AEG-1和SND1均位于HCC细胞的ER膜上。都是AEG-1 SND1 激活 NF-κB。 ER 锚定的 AEG-1 作为上游信号分子的平台 NF-κB 通路因此在 NF-κB 激活中发挥重要作用。 SND1的激活机制 NF-κB 未知。我们假设在转化的肝细胞中，AEG-1 从细胞核转位 锚定到 ER 膜上，招募 SND1，两者合作通过调节促进 HCC RISC 中 mRNA 的转录后调控、膜蛋白的翻译调控和激活 NF-κB、AEG-1 和 SND1 需要彼此才能发挥最佳功能，并且可能不会发挥致癌活性 单独抑制 AEG-1 和 SND1 或联合抑制 AEG-1 和 SND1 可能是 HCC 的有效治疗策略。 实验旨在使用新颖的小鼠模型和有针对性的方法来质疑这些假设 纳米复合物为 AEG-1 和 SND1 提供 siRNA。该提案将有助于我们的长期目标 确定调节 HCC 发病机制的关键参与者，并将这些知识转化为新型药物的开发 以及有效的靶向治疗。该提案的直接目标是深入了解 AEG-1 和 SND1 促进 HCC 的分子机制并评估组合策略 在小鼠模型中抑制 AEG-1 和 SND1 作为潜在的治疗方法。因此，该提案既有 机制和治疗意义和创新。成功完成拟议的研究将 建立开发治疗方法的新目标，并为目标方案提供临床前证据。