Design of Fe2+ and H2O2 Induced Proximity Functionalized Imaging Probes for the Control of Cellular Functions

用于控制细胞功能的 Fe2 和 H2O2 诱导接近功能化成像探针的设计

• 批准号: 9918434
• 负责人: WEI WANG
• 金额: $ 28.78万
• 依托单位: UNIVERSITY OF ARIZONA
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-05-01 至 2023-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9918434
• 关键词: Abscisic Acid; Address; Biological; Biological Process; Biomedical Research; Biotechnology; Brain-Derived Neurotrophic Factor; Buffers; Cell Death; Cell Therapy; Cell physiology; Cells; Chemicals; Complex; Cues; Detection; Disease; Engineering; Environment; Event; Fluorescence Microscopy; Functional Imaging; Future; Genes; Genetic; Gibberellins; High Pressure Liquid Chromatography; Hydrogen Peroxide; Immune response; Knowledge; Logic; Luciferases; Mediating; Mediator of activation protein; Methods; Microscopy; Molecular Biology; Mutation; Nature; Neurodegenerative Disorders; Neurons; Noise; Oxidative Stress; Pathway interactions; Production; Proteins; Reporting; Research; Research Proposals; Series; Signal Pathway; Signal Transduction; Signaling Molecule; Site; Specificity; Stimulus; Technology; Testing; Therapeutic; Toxic effect; Translations; biological systems; design; gene therapy; imaging probe; improved; innovation; new technology; novel; plasmid DNA; programs; protective effect; sensor; side effect; spatiotemporal; stability testing; synthetic biology; theranostics; therapeutic protein; tool

Project Summary Gene/cell therapies finally come of age thanks to the fundamental advances made by innovative and improved biotechnologies. Nonetheless, many challenges particularly toxicity and undesired/uncontrolled immune response remain. This research proposal addresses these concerns and represents a critical first step toward developing novel, more effective cell/gene therapy. Toward this end, we propose a novel synthetic biology technology termed “Environment Stimuli-Induced Proximity (ESIP)” to spatiotemporally manipulate cellular functions. The new strategy can overcome the difficulty of the established chemically induced proximity (CIP) method to mediate the translation of endogenous cellular signals into tailored cellular functions with spatiotemporal precision. To demonstrate the feasibility, we propose to develop fluorescent and chemiluminescent Fe(II) and H2O2 responsive ESIP chemical inducers because Fe(II) and H2O2 are important signal molecules, which are associated with numerous biological functions and diseases, and gibberellin (GA) and abscisic acid (ABA) are established chemical inducers with low toxicity. Specifically, we will design, synthesize, test and optimize Fe2+ and H2O2-responsive fluorescent and chemiluminescent ESIP inducers (Specific Aims 1-2) and test them in cells and construct ESIP-mediated “AND” Boolean logic gates to control the conditional production of AD therapeutic proteins (Specific Aim 3). These studies will prove that these `smart' theranostic probes possess the ability to sense endogenous Fe(II) and H2O2 specifically, and the ability to trigger the release of original bioactive chemical inducers GA and ABA, which then induce downstream biological functions such as luciferase expression and the production of neuron protective BDNF proteins. The technology will ultimately be transformed into entirely new cell/gene therapies for disease treatment with significantly reduced side effects. It is expected that this powerful and general strategy can be integrated with a variety of existing synthetic biology molecular parts and tools to build new cellular genetic and signaling circuitries to generate new functions as toolbox for biomedical research and as therapeutics for various disease treatment.

项目概要 由于创新和技术取得的根本性进展，基因/细胞疗法终于成熟了。 改进的生物技术。尽管如此，仍然存在许多挑战，特别是毒性和不期望/不受控制的 免疫反应依然存在。这项研究提案解决了这些问题，并代表了关键的第一步 致力于开发新颖、更有效的细胞/基因疗法。为此，我们提出了一种新型合成 称为“环境刺激诱导接近（ESIP）”的生物技术来进行时空操纵 细胞功能。新策略可以克服建立化学诱导接近的困难 （CIP）方法介导内源性细胞信号转化为定制的细胞功能 时空精度。为了证明可行性，我们建议开发荧光和 化学发光 Fe(II) 和 H2O2 响应 ESIP 化学诱导剂，因为 Fe(II) 和 H2O2 很重要 与多种生物功能和疾病相关的信号分子和赤霉素 (GA) 和脱落酸（ABA）是已确定的低毒性化学诱导剂。具体来说，我们将设计， 合成、测试和优化 Fe2+ 和 H2O2 响应荧光和化学发光 ESIP 诱导剂 （具体目标1-2）并在细胞中测试它们并构建ESIP介导的“AND”布尔逻辑门来控制 有条件生产 AD 治疗蛋白（具体目标 3）。这些研究将证明这些“聪明” 治疗诊断探针具有特异性感应内源性 Fe(II) 和 H2O2 的能力，并且能够触发 释放原始生物活性化学诱导剂 GA 和 ABA，然后诱导下游生物活性 荧光素酶表达和神经元保护性 BDNF 蛋白的产生等功能。技术 最终将转化为全新的细胞/基因疗法，用于疾病治疗，具有显着的效果 减少副作用。预计这种强大而通用的策略可以与多种集成 现有的合成生物学分子部件和工具，用于构建新的细胞遗传和信号传导电路 产生新的功能作为生物医学研究的工具箱和各种疾病治疗的疗法。