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Live-cell Activity Architecture in Cancer

癌症中的活细胞活性结构

基本信息

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
项目类别：
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

项目摘要

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：细胞机械的基本调节是通过高度动态的信号网络来实现的分子，当失调时，允许癌细胞误解或忽略通常告诉细胞的信号，停止分裂或开始凋亡，导致不受控制的肿瘤生长。在过去的18年里，我的实验室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最前沿。我们已经开发了使能技术，并将时空监管确立为基本范例在细胞信号传导中，发现它的改变导致细胞生长失控。这项由NCI R35支持的研究计划旨在建立一个新的概念框架，具体了解分子活动的细胞组织。我们假设细胞生化活动在空间上被组织成“活动结构”和失调的驱动分子可以重新组织和重新构建这种活动架构，导致失去对细胞生长，分裂，与死在过去的6年里，我们发表了51篇同行评议的文章，并在以下方面取得了重大进展：建立这个框架。我们开发了一流的蛋白质相互作用成像技术以及在分子长度尺度上的活细胞中的酶活性和第一个激酶生物传感器，清醒小鼠的分辨率成像，这为生物化学活动结构提供了证据在不同的尺度上。我们还取得了突破性的发现，蛋白激酶A的调节亚基 (PKA)，RIα，经历液-液相分离（LLPS）以使动态缓冲和空间分离成为可能。普遍存在的第二信使cAMP的区室化，为长期存在的问题我们进一步发现，纤维板层癌（FLC）中的致癌融合可有效抑制RIα， LLPS并诱导异常cAMP信号传导，这导致细胞增殖和细胞转化增加。我们还发现了Ras/ERK通路的新调控机制，并开发了一种新型Ras生物传感器。在建议的研究，我们将有三个重点。首先，我们会发展创新科技，包括超分辨率活动成像，以照亮不同尺度的生化活动架构。其次，我们将阐明cAMP-PKA活性结构紊乱是如何导致肿瘤发生的。 FLC，并进一步发现新的，癌症相关的生物分子缩合物。第三，我们将调查 ERK的时空调节，这是至关重要的生理功能，并确定脆弱的胰腺癌是一种致命的癌症，对Ras-ERK通路上瘾我们还将促进新疗法的开发，用于评估Ras抑制剂和测量靶接合的新测定。

项目成果

期刊论文数量（27）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

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

DOI：
10.1038/s41589-020-00660-y
发表时间：
2021-01
期刊：
Nature chemical biology
影响因子：
14.8
作者：
Zhang JF;Liu B;Hong I;Mo A;Roth RH;Tenner B;Lin W;Zhang JZ;Molina RS;Drobizhev M;Hughes TE;Tian L;Huganir RL;Mehta S;Zhang J
通讯作者：
Zhang J

Fourier Optical Spin Splitting Microscopy.

DOI：
10.1103/physrevlett.129.020801
发表时间：
2022-07
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：
Junxiao Zhou;Qianyi Wu;Junxiang Zhao;Clara Posner;M. Lei;Guanghao Chen;Jin Zhang;Zhaowei Liu

Observing the Assembly of Protein Complexes in Living Eukaryotic Cells in Super-Resolution Using refSOFI.

使用 refSOFI 在超分辨率下观察活体真核细胞中蛋白质复合物的组装。