Sonomagnetic Imaging and Sonomechanical Control of Biological Processes in Deep Tissues

深层组织生物过程的声磁成像和声机械控制

• 批准号: 9750745
• 负责人: George J Lu
• 金额: $ 9.02万
• 依托单位: CALIFORNIA INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-08-01 至 2020-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9750745
• 关键词: Acoustics; Address; Adopted; Anatomy; Animal Model; Animals; Area; Atmosphere; Bacterial Proteins; Basic Science; Biochemical; Biochemistry; Biological Process; Biology; Biomedical Research; Caenorhabditis elegans; Cell Culture Techniques; Cell Size; Cell division; Cell physiology; Cells; Clinical; Collaborations; Coupling; Deposition; Devices; Diagnosis; Disease; Engineering; Escherichia coli; Face; Fungal Proteins; Gases; Gastrointestinal tract structure; Gene Expression; Genetic Engineering; Genotype; Goals; Green Fluorescent Proteins; Human; Hydrophobic Surfaces; Hydrophobicity; Image; Knowledge; Leadership; Light; Location; Magnetic Resonance Imaging; Mammals; Manuscripts; Membrane Proteins; Mentors; Methods; Microbe; Microscope; Modernization; Monitor; Mus; Nanostructures; Nature; Neurons; Opsin; Optics; Organism; Other Genetics; Pathway interactions; Penetration; Permeability; Physiologic pulse; Probiotics; Production; Property; Protein Engineering; Proteins; Protocols documentation; Recording of previous events; Research; Research Institute; Research Personnel; Rodent; Signal Pathway; Signal Transduction; Stains; Surface; System; Techniques; Technology; Testing; Therapeutic; Thick; Time; Tissues; Training; Transgenes; Transmission Electron Microscopy; Ultrasonography; Vesicle; Water; Work; animal imaging; base; biological systems; clinical translation; commensal microbes; design; fluorescence imaging; gut microbes; human subject; in vivo; in vivo imaging; instrumentation; interest; invention; magnetic field; microbiome; microscopic imaging; misfolded protein; monolayer; nanometer; new technology; next generation; novel; optical imaging; optogenetics; particle; post-doctoral training; pressure; probiotic therapy; programs; research and development; skills; spatiotemporal; symposium; synthetic biology; therapy development; tool

Project Summary / Abstract New ways to observe and manipulate cellular function have revolutionized our understanding of biology. Such methods have undergone multiple paradigm shifts in history, from Hooke's microscope and Cajal's staining of neurons, to modern fluorescent imaging based on green fluorescent protein (GFP) and opsin-based optogenetics. These modern genetically encoded methods, defined by their use of protein-based agents that can be expressed by cells, provide key capabilities such as cell-specific targeted expression, continued production in dividing cells, and coupling with state-of-the-art genetic engineering methods. However, as most of these protein tools rely on optical interactions, they are fundamentally limited by the ~ 1 mm penetration depth of light into opaque tissue. As the objects of study increase in size from cell cultures and translucent organisms such as C. elegans, via small rodents, to human beings, this limitation becomes increasingly severe. To overcome this technological challenge, I aim to develop next-generation methods that are both genetically encoded and able to communicate with deeply penetrant forms of energy. In particular, I will leverage the unique physical and biochemical properties of gas vesicles (GVs), a class of gas-filled protein- only nanostructures discovered in certain photosynthetic microbes. In Aim 1, a new method, “sonomagnetic imaging” (SMI), will be developed, which takes advantage of GVs' dual ability to induce contrast for magnetic resonance imaging (MRI) and interact with ultrasound pulses for selective erasing of such contrast. In parallel, a new method to control gene expression, sonomechanical control (SMC), will be developed in Aim 2 wherein ultrasound pulses can collapse GVs and trigger signaling pathways to activate gene expression at high spatial precision and low energy deposition. In Aim 3, I will integrate these novel methods to the task of engineering spatiotemporally trackable and controllable mammalian gut microbes. The completion of these aims will establish a platform for designing probiotics that can be monitored for their function and controlled externally by clinicians to deliver therapies at precise locations and times. Furthermore, the invention of these technologies will stimulate broad interest in other biomedical research that requires sensitive imaging and control of cellular function in deep-lying tissues. The proposal also describes research expertise training, conference attendance and the acquisition of leadership skills that, altogether, will prepare me as a competitive candidate to establish an independent research program at a major research institute and stimulate advances towards my long-term goal of developing imaging and control technologies to study intact biological systems.

项目概要/摘要 观察和操纵细胞功能的新方法彻底改变了我们对生物学的理解。 从胡克的显微镜到卡哈尔的显微镜，这些方法在历史上经历了多次范式转变。 神经元染色，基于绿色荧光蛋白 (GFP) 和视蛋白的现代荧光成像 光遗传学。这些现代基因编码方法通过使用基于蛋白质的试剂来定义 可以由细胞表达，提供细胞特异性靶向表达等关键能力，续 在分裂细胞中生产，并与最先进的基因工程方法相结合。然而，正如大多数 这些蛋白质工具依赖于光学相互作用，它们从根本上受到 ~ 1 毫米穿透力的限制 光进入不透明组织的深度。随着细胞培养物和半透明研究对象尺寸的增加 从线虫等生物体，通过小型啮齿动物，到人类，这种限制变得越来越明显 严重。为了克服这一技术挑战，我的目标是开发下一代方法 基因编码并能够与深层渗透的能量形式进行交流。特别是，我将 利用气体囊泡 (GV) 的独特物理和生化特性，气体囊泡是一类充满气体的蛋白质 仅在某些光合微生物中发现的纳米结构。在目标 1 中，一种新方法“声磁 将开发“成像”（SMI），它利用 GV 的双重能力来诱导磁性对比 磁共振成像（MRI）并与超声波脉冲相互作用以选择性地消除这种对比度。并联， 目标 2 将开发一种控制基因表达的新方法，即声机械控制 (SMC)，其中 超声波脉冲可以瓦解 GV 并触发信号通路，从而在高空间激活基因表达 精度和低能量沉积。在目标 3 中，我将把这些新颖的方法整合到工程任务中 时空可追踪和可控的哺乳动物肠道微生物。这些目标的完成将 建立一个设计益生菌的平台，可以监测其功能并通过外部控制 临床医生可以在精确的地点和时间提供治疗。此外，这些技术的发明 将激发人们对其他需要敏感成像和细胞控制的生物医学研究的广泛兴趣 在深层组织中发挥作用。该提案还描述了研究专业知识培训、会议出席 以及领导技能的获得，这些技能总的来说将使我成为一名有竞争力的候选人，建立 一个主要研究机构的独立研究项目，并刺激我的长期进步 开发成像和控制技术来研究完整的生物系统的目标。