Regulation of oncogenic Akt ubiquitination and activation by diverse mechanisms in cancer

癌症中多种机制对致癌 Akt 泛素化和激活的调节

• 批准号: 10907082
• 负责人: Hui-Kuan Lin
• 金额: $ 48.57万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-01 至 2026-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10907082
• 关键词: Address; Automobile Driving; Biochemical; Biological; Biological Process; Cancer Intervention; Cell Proliferation; Complex; Development; Dose; Event; Genetic; Goals; Growth Factor; Human; Insulin-Like Growth Factor I; Knock-in Mouse; Knowledge; Ligase; Link; Malignant Neoplasms; Mediating; Membrane; Metabolism; Methylation; Methyltransferase; Modeling; Modification; Mus; Oncogenic; Organoids; PIK3CG gene; PRKDC gene; Patients; Phosphorylation; Phosphotransferases; Play; Post-Translational Protein Processing; Process; Proteomics; Regulation; Research; Role; SETDB1 gene; Signal Pathway; Signal Transduction; TRAF6 gene; Technology; Treatment Factor; Ubiquitination; Xenograft procedure; cancer initiation; cancer therapy; druggable target; frontier; innovation; insight; mouse model; novel; pharmacologic; recruit; side effect; small molecule inhibitor; therapeutic target; tumor; tumor progression; tumor xenograft; tumorigenesis; ubiquitin-protein ligase

Summary The PI3K/Akt signaling is one of most important oncogenic events in human cancers. It regulates many aspects of biological functions including cell proliferation, survival, and metabolism important for cancer initiation and progression. While extensive efforts have been made in the last three decades to understand the downstream effectors responsible for the biological and oncogenic processes regulated by PI3K/Akt signaling, the upstream signals mediating PI3K/Akt signaling activation upon diverse growth factor stimulation is not well understood. Understanding and defining the upstream mechanisms responsible for PI3K/Akt signaling activation will not only provide new insight into how PI3K/Akt signaling activation is orchestrated, but also offer novel paradigms and therapeutic targets for cancer intervention. Although it has been well established that PIP3 is critical for the membrane recruitment and subsequent activation of Akt, our recent studies provide the evidence that Akt undergoes methylation and subsequent non-proteolytic K63-linked ubiquitination, which are crucial for Akt membrane recruitment and subsequent phosphorylation and activation upon stimulation with diverse growth factors, opening up a new frontier for Akt signaling regulation. Of note, we identified SETDB1 as a methyltransferase for Akt K64 methylation and TRAF6 ligase as an upstream E3 ligase triggering K63-linked ubiquitination and activation of Akt, and these events are required for cancer progression. However, the outstanding questions remained to be addressed are how SETDB1 and TRAF6 are activated or recruited to the Akt complex upon growth factor treatment to trigger Akt methylation and subsequent Akt ubiquitination and activation, thus promoting oncogenic processes. The goal of this study is to dissect the upstream regulatory mechanisms by which growth factors activate and recruit SETDB1 and TRAF6 ligase to Akt complex to elicit Akt methylation and subsequent Akt ubiquitination, define the mechanism by which Akt ubiquitination facilitates Akt membrane localization and activation, and finally explore the role of these regulatory modes in cancer development and develop small molecule inhibitors targeting these regulatory mechanisms. Our preliminary results revealed that SETDB1 and TRAF6 undergo novel posttranslational modifications, which are crucial for methylation, ubiquitination and activation of Akt by growth factors and oncogenic activity. We hypothesized that SETDB1 and TRAF6 undergo the novel posttranslational modification upon growth factor treatment, which recruits SETDB1 to the Akt complex and activates TRAF6 E3 ligase to facilitate Akt methylation and subsequent Akt ubiquitination and activation, thus leading to tumorigenesis. Our innovative hypothesis has been formulated based on our preliminary results and prior research. We proposed three specific aims to validate this provocative and paradigm-shifting concept using cutting-edge technologies including xenograft, organoids from genetic mouse tumor models and patient derived tumors, patient-derived models (PDX), knockin mouse models, genetic mouse tumor models and pharmacological approaches. This application is significant, therefore, because it is expected to provide the knowledge needed to develop pharmacologic strategies that will allow concurring cancers with aberrant PI3K/Akt activation. Our study will open up a new avenue for PI3K/Akt signaling regulation, but also offer new concepts and strategies for cancer targeting.

概括 PI3K/Akt 信号传导是人类癌症中最重要的致癌事件之一。它规范了很多方面 生物功能，包括细胞增殖、存活和代谢，对于癌症的发生和发展非常重要 进展。尽管在过去三十年中人们做出了大量努力来了解下游 负责受 PI3K/Akt 信号传导（上游）调节的生物和致癌过程的效应器 在不同生长因子刺激下介导 PI3K/Akt 信号激活的信号尚不清楚。 理解和定义负责 PI3K/Akt 信号激活的上游机制不会 不仅提供了关于如何协调 PI3K/Akt 信号激活的新见解，而且还提供了新颖的范例 和癌症干预的治疗目标。尽管 PIP3 对于 Akt 的膜募集和随后的激活，我们最近的研究提供了证据 经历甲基化和随后的非蛋白水解 K63 连接泛素化，这对于 Akt 至关重要 不同生长刺激下的膜募集和随后的磷酸化和激活 因素，为 Akt 信号调控开辟了新领域。值得注意的是，我们将 SETDB1 确定为 用于 Akt K64 甲基化的甲基转移酶和 TRAF6 连接酶作为触发 K63 连接的上游 E3 连接酶 Akt 的泛素化和激活，这些事件是癌症进展所必需的。然而， 仍有待解决的突出问题是如何激活 SETDB1 和 TRAF6 或将其招募到 Akt 复合物经生长因子处理触发 Akt 甲基化和随后的 Akt 泛素化 激活，从而促进致癌过程。本研究的目的是剖析上游监管 生长因子激活并招募 SETDB1 和 TRAF6 连接酶至 Akt 复合物以引发的机制 Akt 甲基化和随后的 Akt 泛素化定义了 Akt 泛素化的机制 促进Akt膜定位和激活，最终探讨这些调节模式在Akt膜中的作用 癌症的发展并开发针对这些调节机制的小分子抑制剂。我们的 初步结果显示 SETDB1 和 TRAF6 经历了新颖的翻译后修饰，即 对于生长因子和致癌活性的 Akt 甲基化、泛素化和激活至关重要。我们 假设 SETDB1 和 TRAF6 经历了生长因子的新型翻译后修饰 治疗，将 SETDB1 招募到 Akt 复合物并激活 TRAF6 E3 连接酶以促进 Akt 甲基化以及随后的 Akt 泛素化和激活，从而导致肿瘤发生。我们的创新 假设是根据我们的初步结果和先前的研究制定的。我们提出了三个 具体目标是利用尖端技术验证这一具有挑衅性和范式转变的概念 包括异种移植物、来自遗传性小鼠肿瘤模型和患者源性肿瘤的类器官、患者源性肿瘤 模型（PDX）、敲入小鼠模型、遗传小鼠肿瘤模型和药理学方法。这 因此，应用程序很重要，因为它有望提供开发所需的知识 允许癌症同时发生异常 PI3K/Akt 激活的药理学策略。我们的研究将 开辟了PI3K/Akt信号调节的新途径，也为癌症提供了新的概念和策略 瞄准。

项目成果

Protocol to analyze the role of various metabolites in impacting global RNA 5-methylcytosine levels in cultured cells by dot blot.

• DOI: 10.1016/j.xpro.2023.102815
• 发表时间: 2024-03-15
• 期刊: STAR PROTOCOLS
• 作者: Chen, Tingjin;Lin, Hui-Kuan
• 通讯作者: Lin, Hui-Kuan

Identification of myo-inositol-binding proteins by using the biotin pull-down strategy in cultured cells.

• DOI: 10.1016/j.xpro.2022.101385
• 发表时间: 2022-06-17
• 期刊: STAR PROTOCOLS
• 作者: Hsu, Che-Chia;Xu, Zhi-Gang;Lei, Jie;Chen, Zhong-Zhu;Li, Hong-Yu;Lin, Hui-Kuan
• 通讯作者: Lin, Hui-Kuan

UHRF1: a novel metabolic guardian restricting AMPK activity.

UHRF1：一种限制 AMPK 活性的新型代谢卫士。

• DOI: 10.1038/s41422-021-00589-4
• 发表时间: 2022
• 期刊: Cell research
• 影响因子: 44.1
• 作者: Hsu,Che-Chia;Tsai,Yau-Sheng;Lin,Hui-Kuan
• 通讯作者: Lin,Hui-Kuan