Sonogenetic Remote Control of Cellular Function

细胞功能的声遗传学远程控制

• 批准号: 10261864
• 负责人: Mikhail Shapiro
• 金额: $ 117.25万
• 依托单位: CALIFORNIA INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-30 至 2026-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10261864
• 关键词: Acoustics; Anatomy; Animals; Biological; Biological Phenomena; Biology; Biosensor; Brain region; Cell Therapy; Cell physiology; Cells; Communication; Deposition; Development; Engineering; Fluorescence Microscopy; Focused Ultrasound; Gastrointestinal tract structure; Gene Expression; Goals; Human body; Image; Immune; Immunotherapy; Investigation; Light; Location; Mechanics; Medical Imaging; Medical Technology; Methods; Modality; Neurons; Neurosciences; Operative Surgical Procedures; Optics; Pathway interactions; Penetration; Physics; Process; Property; Proteins; Reporter Genes; Research; Research Personnel; Signal Transduction; Structure; Techniques; Technology; Time; Tissues; Travel; Tumor-infiltrating immune cells; Ultrasonography; Visible Radiation; Work; biological systems; cellular imaging; in vivo; mechanical force; microbial colonization; molecular imaging; neuroregulation; optogenetics; receptor; remote control; sound; tool; tumor

SUMMARY The discovery and development of fluorescent proteins and optogenetics revolutionized biology by making it possible to image and control specific cellular processes with visible light. While these tools have enabled countless biological discoveries, the poor penetration of light into living tissue makes it difficult to use optical techniques in intact animals. As a result, biological phenomena ranging from the signaling of neurons in deep- brain regions, to the infiltration of immune cells into tumors, to the microbial colonization of the GI tract, are challenging to study within their natural in vivo context. If instead of light it were possible to visualize and manipulate cellular function using a more penetrant form of energy such as ultrasound, this would open previously inaccessible domains of in vivo biology to direct investigation. In addition, it would enhance the development of cell-based therapies by allowing cellular agents to be seen and controlled after administration into the human body. The physics of ultrasound make it an ideal modality for deep-tissue cellular communication. Sound waves in the MHz range are weakly scattered by tissue and can therefore penetrate several cm into the body. With wavelengths on the order of 100 µm and travel times < 1 ms, ultrasound can access many key structures and processes. When focused, sound waves can deliver mechanical and thermal energy to precise anatomical locations. These properties have already made ultrasound one of the world’s most widely used technologies for medical imaging and non-invasive surgery. However, the potential of ultrasound to serve as a tool for cellular imaging and control has been relatively untapped due to a lack of methods to connect it to the function of specific cells and biomolecules. In previous work, the Shapiro lab has pioneered the use of ultrasound in cellular and molecular imaging by developing the first acoustic reporter genes and biosensors for ultrasound, aiming to “do for ultrasound what fluorescent proteins have done for fluorescence microscopy”. The major goal of our proposed new research direction is to “do for ultrasound what optogenetics has done for light” by giving sound waves the ability to control specific cellular functions such as neuronal excitation, gene expression and intracellular signaling in vivo. The basic principle of our approach is to (1) use focused ultrasound to deposit acoustic energy at a specific location in tissue, (2) use genetically encoded “acoustic antennae” to convert this energy into local mechanical force, and (3) use this force to actuate mechanosensitive receptors to produce specific cellular signals. We will implement this approach in neurons and immune cells to enable unique neuroscience and cell therapy applications. If successful, this work will help establish the new field of sonogenetics by providing researchers and clinicians with the unprecedented ability to “point and click” on cells deep within the body and tell them what to do.

概括 荧光蛋白和光遗传学的发现和发展彻底改变了生物学 虽然这些工具已经启用，但可以用可见光对特定的细胞过程进行成像和控制。 无数的生物学发现，光对活体组织的渗透性较差，使得光学技术难以使用 结果，从深部神经元信号传导的生物现象。 大脑区域、免疫细胞浸润到肿瘤、胃肠道微生物定植，这些都是 如果可以不用光来进行可视化和观察，那么在它们的自然体内环境中进行研究就具有挑战性。 使用更具渗透性的能量形式（例如超声波）来操纵细胞功能，这将打开 此外，它将增强以前无法进入的体内生物学领域的直接研究。 通过允许在给药后观察和控制细胞制剂来开发基于细胞的疗法 超声波的物理原理使其成为深层组织细胞通讯的理想方式。 MHz范围内的声波被组织微弱散射，因此可以穿透几厘米进入组织 超声波的波长约为 100 µm，传播时间 < 1 ms，可以访问许多按键。 当聚焦时，声波可以精确地传递机械能和热能。 这些特性已经使超声波成为世界上使用最广泛的技术之一。 然而，超声波作为医疗成像和非侵入性手术的潜力。 由于缺乏将其连接到细胞的方法，用于细胞成像和控制的工具相对尚未开发。 在之前的工作中，夏皮罗实验室率先使用了超声波。 通过开发第一个声学报告基因和超声波生物传感器，在细胞和分子成像领域， 旨在“为超声波做荧光蛋白为荧光显微镜所做的事情”。 我们提出的新研究方向是“为超声波做光遗传学为光所做的事情” 声波控制特定细胞功能的能力，例如神经元兴奋、基因表达和 我们的方法的基本原理是（1）使用聚焦超声沉积。 组织中特定位置的声能，(2) 使用基因编码的“声波天线”将其转换为 将能量转化为局部机械力，并且（3）利用该力来驱动机械敏感受体以产生 我们将在神经元和免疫细胞中实施这种方法，以实现独特的细胞信号。 如果成功，这项工作将有助于建立神经科学和细胞治疗应用的新领域。 声遗传学为研究人员和反叛者提供前所未有的“指向并点击”细胞的能力 深入身体并告诉他们该怎么做。