Sonogenetic Remote Control of Cellular Function

总结 荧光蛋白和光遗传学的发现和发展使生物学发生了革命性的变化， 可以用可见光成像和控制特定的细胞过程。虽然这些工具使 无数的生物学发现，光对活组织的渗透性差，使得很难使用光学 技术在完整的动物。因此，生物现象，从深层神经元的信号， 大脑区域，免疫细胞浸润到肿瘤中，到胃肠道的微生物定植， 在其自然的体内环境中进行研究具有挑战性。如果不是光，而是视觉， 操纵细胞功能使用更渗透形式的能量，如超声波，这将打开 以前无法进入的领域在体内生物学直接调查。此外，它还将增强 通过在给药后观察和控制细胞因子来开发基于细胞的疗法 植入人体超声的物理特性使其成为深层组织细胞通信的理想模式。 MHz范围内的声波被组织微弱地散射，因此可以穿透几厘米进入组织。 身体超声波的波长约为100 µm，传播时间< 1 ms， 结构和工艺。当聚焦时，声波可以将机械能和热能精确地传递到 解剖位置。这些特性已经使超声波成为世界上应用最广泛的技术之一 医学成像和非侵入性手术技术。然而，超声波作为一种 细胞成像和控制的工具一直相对未开发，由于缺乏方法将其连接到 特定细胞和生物分子的功能。在之前的工作中，夏皮罗实验室率先使用超声波 在细胞和分子成像中，通过开发第一个声学报告基因和用于超声的生物传感器， 其目标是“为超声波做荧光蛋白为荧光显微镜做的事情”。的主要目标 我们提出的一个新的研究方向是“为超声做光遗传学为光做的事情”， 声波控制特定细胞功能的能力，如神经元兴奋，基因表达和 体内细胞内信号传导。我们的方法的基本原理是（1）使用聚焦超声存款 组织中特定位置的声能，（2）使用基因编码的“声天线”将其转换为 能量转化为局部机械力，以及（3）使用该力来驱动机械敏感受体以产生 特定的细胞信号。我们将在神经元和免疫细胞中实施这种方法， 神经科学和细胞治疗应用。如果成功，这项工作将有助于建立新的领域， 通过为研究人员和临床医生提供前所未有的“指向和点击”细胞的能力， 告诉他们该怎么做

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.