Sonomagnetic Imaging and Sonomechanical Control of Biological Processes in Deep Tissues

项目摘要/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。 这种方法在历史上经历了多次范式转变，从胡克的显微镜到卡哈尔的 神经元染色，到基于绿色荧光蛋白(GFP)和视蛋白的现代荧光成像 光遗传学。这些现代的基因编码方法，通过使用基于蛋白质的试剂来定义 可以通过细胞表达，提供细胞特异性靶向表达等关键功能，续 在分裂细胞中进行生产，并结合最先进的基因工程方法。然而，正如大多数人一样 在这些依赖于光学相互作用的蛋白质工具中，它们从根本上受到~1 mm穿透的限制 光线进入不透明组织的深度。随着研究对象从细胞培养和半透明的尺寸增加 例如线虫等生物，通过小型啮齿动物传播给人类，这种限制变得越来越多 很严重。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，并能够与深度渗透的能量形式进行交流。尤其是，我会 利用气泡(GV)的独特物理和生化特性，气泡是一类充满气体的蛋白质- 只有在某些光合作用微生物中发现的纳米结构。在目标1中，采用了一种新的方法--“声磁 成像“(SMI)将被开发出来，它利用GV的双重能力来产生对比度以获得磁 磁共振成像(MRI)，并与超声脉冲相互作用，选择性地消除这种对比。同时， 在目标2中，将开发一种控制基因表达的新方法，即声机械控制(SMC)，其中 超声脉冲可以使GV崩溃，并触发信号通路以激活高空间基因表达 精度高，能量沉积低。在目标3中，我将把这些新方法集成到工程任务中 时空可追踪和可控制的哺乳动物肠道微生物。这些目标的实现将 建立一个可以监测其功能并由外部控制的益生菌设计平台 临床医生在准确的地点和时间提供治疗。此外，这些技术的发明 将激发人们对其他生物医学研究的广泛兴趣，这些研究需要灵敏的成像和细胞控制 在深层组织中发挥作用。该提案还描述了研究专门知识培训、会议出席情况 以及获得领导技能，这些技能将使我成为一名有竞争力的候选人，以建立 在一家主要研究机构的独立研究计划，并刺激我的长期发展 开发成像和控制技术以研究完整的生物系统的目标。

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.