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

项目摘要 基因/细胞疗法终于成熟了，这要归功于创新和 改进的生物技术。尽管如此，许多挑战，特别是毒性和不希望的/不受控制的 免疫反应仍然存在。这项研究提案解决了这些问题，是关键的第一步。 开发新的、更有效的细胞/基因疗法。为此，我们提出了一种新的合成 生物技术称为"环境刺激诱导邻近（ESIP）"，以时空操纵 细胞功能。新策略可以克服已建立的化学诱导邻近的困难 (CIP)介导内源性细胞信号翻译成定制细胞功能的方法， 时空精度为了证明可行性，我们建议开发荧光和 荧光Fe（II）和H2O2响应的ESIP化学诱导剂，因为Fe（II）和H2O2是重要的 与多种生物学功能和疾病相关的信号分子，以及赤霉素（GA） 和脱落酸（ABA）是公认的低毒化学诱导剂。具体来说，我们将设计， Fe~（2+）和H_2O_2响应性荧光和荧光ESIP诱导剂合成、测试和优化 （具体目的1 - 2）并在细胞中测试它们，并构建ESIP介导的"与"布尔逻辑门以控制细胞的增殖。 AD治疗性蛋白质的条件性产生（特异性目标3）。这些研究将证明，这些"聪明"的 治疗诊断探针具有特异性地感测内源性Fe（II）和H2O2的能力，以及触发 原始生物活性化学诱导物GA和ABA的释放，其然后诱导下游生物活性物质的释放， 功能如荧光素酶表达和神经元保护性BDNF蛋白的产生。技术 最终将转化为全新的细胞/基因疗法，用于疾病治疗， 减少副作用。预计这一强有力的通用战略可以与各种 现有的合成生物学分子部件和工具，以建立新的细胞遗传和信号电路， 作为生物医学研究的工具箱和各种疾病治疗的治疗剂产生新的功能。

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.