Live-cell Activity Architecture in Cancer

癌症中的活细胞活性结构

• 批准号: 10673027
• 负责人: Jin Zhang
• 金额: $ 92.9万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-08-01 至 2029-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10673027
• 关键词: Apoptosis; Architecture; Automobile Driving; Biochemical; Biochemistry; Biological Assay; Biosensor; Buffers; Cell Proliferation; Cells; Cessation of life; Cyclic AMP; Cyclic AMP-Dependent Protein Kinases; Development; Fibrolamellar Hepatocellular Carcinoma; Goals; Image; Imaging technology; Laboratories; Length; Liquid substance; Malignant Neoplasms; Malignant neoplasm of pancreas; Measures; Molecular; Mus; Oncogenic; Pathway interactions; Peer Review; Phase; Phosphotransferases; Physical condensation; Physiological; Protein Subunits; Publishing; Ras Inhibitor; Regulation; Research; Research Support; Second Messenger Systems; Signal Transduction; Signaling Molecule; Suicide; Technology; Therapeutic; anticancer research; awake; cancer cell; cell growth; cell growth regulation; cell transformation; high resolution imaging; innovative technologies; novel; novel therapeutics; programs; protein protein interaction; spatiotemporal; tumor growth; tumorigenesis; ultra high resolution; uncontrolled cell growth

Project summary: Essential regulation of the cellular machinery is achieved by a network of highly dynamic signaling molecules, which, when dysregulated, allow cancer cells to misinterpret or ignore signals that normally tell cells to stop dividing or begin apoptosis, leading to uncontrolled tumor growth. For the last 18 years, my laboratory has been at the forefront of applying a native biochemistry approach to cell signaling and cancer research. We have developed enabling technologies, and established spatiotemporal regulation as a fundamental paradigm in cell signaling, and discovered that its alteration leads to uncontrolled cell growth. This NCI R35-supported research program seeks to establish a new conceptual framework to specifically understand the cellular organization of molecular activities. We hypothesize that cellular biochemical activities are spatially organized into an “activity architecture” and dysregulated driver molecules can re-organize and re-structure this activity architecture, leading to loss of control over cell growth, division and death. In the past 6 years, we published 51 peer-reviewed articles, and made significant advances in establishing this framework. We developed first-in-class technologies for imaging protein-protein interactions and enzymatic activities in living cells at molecular length-scales and first kinase biosensor that achieved high- resolution imaging in awake mice, which have provided evidence for the biochemical activity architecture across different scales. We also made a breakthrough discovery that a regulatory subunit of Protein Kinase A (PKA), RIα, undergoes liquid-liquid phase-separation (LLPS) to enable the dynamic buffering and spatial compartmentalization of a ubiquitous second messenger, cAMP, providing an answer to a long standing question. We further showed that the oncogenic fusion in fibrolamellar carcinoma (FLC) potently inhibits RIα LLPS and induces aberrant cAMP signaling, which leads to increased cell proliferation and cell transformation. We have also discovered novel regulation in the Ras/ERK pathway and developed a novel Ras biosensor. In the proposed research, we will have three focuses. First, we will develop innovative technologies including super-resolution activity imaging to illuminate the biochemical activity architecture across different scales. Secondly, we will elucidate how the disorganized cAMP-PKA activity architecture leads to tumorigenesis in FLC, and further discover novel, cancer-relevant biomolecular condensates. Thirdly, we will investigate the spatiotemporal regulation of ERK that is critical for its physiological functions and identify the vulnerable connections in the re-organized cancer-driving architecture in pancreatic cancer, which is a deadly cancer that is addicted to the Ras-ERK pathway. We will also facilitate the development of new therapeutics by developing novel assays for evaluating Ras inhibitors and measuring target engagement.

项目概要： 细胞机器的基本调节是通过高度动态的信号网络实现的 当这些分子失调时，癌细胞会误解或忽略通常告诉细胞的信号 停止分裂或开始细胞凋亡，导致肿瘤生长失控 在过去 18 年里，我的实验室 我们一直处于将天然生物化学方法应用于细胞信号传导和癌症研究的前沿。 开发了使能技术，并将时空调节作为基本范式 研究细胞信号传导，发现其改变会导致细胞生长失控。 这项 NCI R35 支持的研究计划旨在建立一个新的概念框架 专门了解分子活动的细胞组织。 生化活动在空间上组织成“活动架构”和失调的驱动分子 可以重新组织和重构这种活动架构，导致细胞生长、分裂失去控制 在过去 6 年里，我们发表了 51 篇同行评审文章，并在以下方面取得了重大进展： 我们开发了用于成像蛋白质-蛋白质相互作用的一流技术。 和活细胞中分子长度尺度的酶活性以及第一个实现高 清醒小鼠的高分辨率成像，为生化活性结构提供了证据 我们还取得了突破性的发现，即蛋白激酶 A 的调节亚基。 （PKA）、RIα，经过液-液相分离（LLPS）以实现动态缓冲和空间 普遍存在的第二信使 cAMP 的划分，为长期存在的问题提供了答案 我们进一步表明，纤维板层癌（FLC）中的致癌融合可有效抑制 RIα。 LLPS 会诱导异常的 cAMP 信号传导，从而导致细胞增殖和细胞转化增加。 我们还发现了 Ras/ERK 通路的新调控，并开发了一种新型 Ras 生物传感器。 拟议的研究，我们将有三个重点，第一，我们将开发创新技术，包括。 超分辨率活动成像可阐明不同尺度的生化活动结构。 其次，我们将阐明杂乱的 cAMP-PKA 活性结构如何导致肿瘤发生 FLC，并进一步发现新型的、与癌症相关的生物分子凝聚物。第三，我们将研究这些。 ERK 的时空调节对其生理功能和识别弱势群体至关重要 胰腺癌中重新组织的癌症驱动结构中的联系，这是一种致命的癌症 我们还将通过开发Ras-ERK途径来促进新疗法的开发。 用于评估 Ras 抑制剂和测量靶点参与度的新方法。

项目成果

A genetically encoded near-infrared fluorescent calcium ion indicator.

一种基因编码的近红外荧光钙离子指示剂。

• 发表时间: 2019
• 影响因子: 48
• 作者: Qian, Yong;Piatkevich, Kiryl D;Mc Larney, Benedict;Abdelfattah, Ahmed S;Mehta, Sohum;Murdock, Mitchell H;Gottschalk, Sven;Molina, Rosana S;Zhang, Wei;Chen, Yingche;Wu, Jiahui;Drobizhev, Mikhail;Hughes, Thomas E;Zhang, Jin;Schreiter, Eric R
• 通讯作者: Schreiter, Eric R

Fluorescent Biosensors for Multiplexed Imaging of Phosphoinositide Dynamics.

用于磷酸肌醇动力学多重成像的荧光生物传感器。

• 发表时间: 2020
• 影响因子: 4
• 作者: Hertel, Fabian;Li, Simin;Chen, Mingyuan;Pott, Lutz;Mehta, Sohum;Zhang, Jin
• 通讯作者: Zhang, Jin

Fourier Optical Spin Splitting Microscopy.

傅里叶光学自旋分裂显微镜。

• DOI: 10.1103/physrevlett.129.020801
• 发表时间: 2022-07-05
• 期刊: Physical review letters
• 影响因子: 8.6
• 作者: Junxiao Zhou;Qianyi Wu;Junxiang Zhao;Clara Posner;M. Lei;Guanghao Chen;Jin Zhang;Zhaowei Liu
• 通讯作者: Zhaowei Liu

Single-fluorophore biosensors for sensitive and multiplexed detection of signalling activities.

单荧光团生物传感器，用于信号活动的灵敏和多重检测。

• 发表时间: 2018-10
• 影响因子: 21.3
• 作者: Mehta, Sohum;Zhang, Yong;Roth, Richard H;Zhang, Jin;Mo, Albert;Tenner, Brian;Huganir, Richard L;Zhang, Jin
• 通讯作者: Zhang, Jin

An ultrasensitive biosensor for high-resolution kinase activity imaging in awake mice.

一种超灵敏生物传感器，用于对清醒小鼠进行高分辨率激酶活性成像。

• 发表时间: 2021
• 影响因子: 14.8
• 作者: Zhang, Jin;Liu, Bian;Hong, Ingie;Mo, Albert;Roth, Richard H;Tenner, Brian;Lin, Wei;Zhang, Jason Z;Molina, Rosana S;Drobizhev, Mikhail;Hughes, Thomas E;Tian, Lin;Huganir, Richard L;Mehta, Sohum;Zhang, Jin
• 通讯作者: Zhang, Jin