Elucidating the fundamental principles of cell polarity using synthetic biology

利用合成生物学阐明细胞极性的基本原理

• 批准号: 7275652
• 负责人: ALEXANDER L WATTERS
• 金额: $ 4.68万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-01 至 2009-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7275652
• 关键词: Accounting; Actins; Back; Cell Polarity; Cell physiology; Cells; Characteristics; Data; Development; Engineering; F-Actin; Feedback; Future; Genetic Transcription; Immune response; Individual; Knowledge; Lead; Link; Localized; Malignant Neoplasms; Mammalian Cell; Methods; Modeling; Movement; Myosin ATPase; Neoplasm Metastasis; Property; Protein Overexpression; Proteins; Purpose; Range; Regulation; Relative (related person); Reverse Transcriptase Polymerase Chain Reaction; Set protein; Signal Pathway; Signal Transduction; Structure; System; Takeda brand of pioglitazone hydrochloride; Testing; Therapeutic; Tumor Cell Invasion; Wound Healing; base; cancer cell; cell motility; design; insight; novel; novel therapeutics; polymerization; research study; synthetic biology; synthetic protein; tumor

DESCRIPTION (provided by applicant): Actin-based motility is critical for diverse cellular processes, including development, wound healing, the immune response, and tumor invasion and metastasis. A common property of all motile cells is their ability to polarize - they show distinct asymmetry, with a front containing F-actin protrusive structures and a back containing acto-myosin contractile structures. While most cells possess the ability to generate the individual actin formations underlying motility, there is significant variability in the ability of mammalian cells to coordinate the asymmetric polarity of protrusion and contraction required for directional motility. Objective/Hypotheses: This project will explore two possible fundamental principles behind the variability in cell polarization and motility: 1) non-motile cells express all of the necessary components for motility, but that they are not capable of generating an initial level of activation needed to drive the system to polarity, or 2) non-motile cells are missing the requisite feedback circuits needed for separation of the "front" and the "back". The following Specific Aims will be used to test these hypotheses by attempting to convert non-motile and semi-motile cells into highly motile cells. 1) Overexpressing endogenous proteins or sets of proteins involved in actin regulation, (e.g. co- expression of high levels of antagonistic activators of F-actin and acto-myosin). These proteins will be choosen based on results of RT-PCR experiments used to identify endogenous proteins whose expression correlates with cell motility (e.g. up regulated in highly motile cells relative to semi- and non-motile cells). 2) Introducing synthetic signaling circuits specifically engineered to generate local positive feedback loops for F-actin formation, and long-range cross-inhibition between the signaling pathways responsible for F-actin and acto-myosin. These novel synthetic proteins will be designed to be regulated and localized by specific activators of F-actin and acto-myosin structures using methods developed in the Lim Lab. Relevance: Despite the results of many studies suggesting a link between the malignancy of tumors and the deregulation of actin polymerization, little is understood about how this deregulation occurs. Thus, understanding how non-motile cells can acquire the ability to generate and amplify actin based polarity will provide insight not only into how natural cells normally achieve movement, but also into how non-motile cancer cells gain the ability to metastasize. Such knowledge will be useful for future attempts at controlling metastatic tumors and developing novel therapeutics based on controlling cell motility.

描述（由申请人提供）：基于肌动蛋白的运动对于各种细胞过程至关重要，包括发育，伤口愈合，免疫反应以及肿瘤侵袭和转移。所有运动细胞的共同特性是它们的两极分化的能力 - 它们显示出独特的不对称性，前部包含F-肌动蛋白突出结构和一个含有acto肌球蛋白收缩结构的背面。虽然大多数细胞具有产生单个肌动蛋白形成的能力，但哺乳动物细胞协调方向运动所需的突出和收缩的不对称极性的能力存在显着差异。客观/假设：该项目将探索细胞极化和运动变异性背后的两个可能的基本原理：1）非运动细胞表达运动性运动的所有必要组件，但是它们无法产生将系统驱动到极性所需的初始激活水平，或者2）非摩托车细胞所需的反馈反馈循环所需的反馈循环和前方的前方和'''''''''''''''''''''以下特定目的将用于通过尝试将非运动和半运动细胞转换为高度运动细胞来检验这些假设。 1）过表达参与肌动蛋白调节的内源性蛋白质或一组蛋白质（例如，F-肌动蛋白和肌动蛋白的高水平拮抗激活剂的共同表达）。这些蛋白质将基于用于鉴定内源性蛋白的RT-PCR实验的结果来选择其表达与细胞运动相关的内源性蛋白（例如，在高度运动细胞中相对于半运动细胞与半运动细胞的调节）。 2）引入专门设计的合成信号通路，以生成用于F-肌动蛋白形成的局部正反馈回路，以及负责F-肌动蛋白和肌动蛋白的信号传导途径之间的远程交叉抑制。这些新型的合成蛋白将设计为使用LIM实验室中开发的方法由F-肌动蛋白和肌动蛋白肌球蛋白结构的特定活化剂调节和定位。相关性：尽管许多研究的结果表明，肿瘤的恶性肿瘤与肌动蛋白聚合失调之间存在联系，但几乎没有理解这种放松管制。因此，了解非运动细胞如何获得产生和扩增基于肌动蛋白的极性的能力，不仅会洞悉天然细胞通常如何实现运动，还可以介绍非运动癌细胞如何获得转移的能力。这种知识将对未来控制转移性肿瘤和基于控制细胞运动的新型治疗剂的尝试很有用。