Acoustically targeted molecular control of cell type specific neural circuits in non-human primates

非人类灵长类动物细胞类型特异性神经回路的声学靶向分子控制

• 批准号: 9804641
• 负责人: Mikhail Shapiro
• 金额: $ 116.22万
• 依托单位: CALIFORNIA INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-08-15 至 2022-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9804641
• 关键词: Acoustics; Animal Model; Animals; Area; BRAIN initiative; Behavioral; Bioavailable; Blood - brain barrier anatomy; Brain; Brain region; Cells; Clinical; Complex; Development; Directed Molecular Evolution; Disease; Dose; Engineering; Evolution; Face; Focused Ultrasound; Functional Imaging; Future; Gene Delivery; Genetic; Goals; Hippocampus (Brain); Histologic; Human; Immunologics; Ligands; Location; Macaca; Magnetic Resonance Imaging; Mediating; Memory; Methods; Molecular Target; Monitor; Monkeys; Mus; Names; Nature; Neurons; Neurosciences; Oral; Organism; Patients; Performance; Pharmacology; Phase; Primates; Procedures; Reagent; Safety; Serotyping; Specificity; System; Techniques; Technology; Testing; Transfection; Translating; Translations; Ultrasonics; Ultrasonography; Viral; Viral Vector; Virus; Work; XCL1 gene; adeno-associated viral vector; behavioral study; cell type; cranium; design; designer receptors exclusively activated by designer drugs; excitatory neuron; experimental study; image guided; improved; in vivo; innovation; intersectionality; millimeter; neural circuit; neuropsychiatric disorder; neuroregulation; non-invasive imaging; nonhuman primate; optogenetics; pressure; receptor; relating to nervous system; small molecule; success; translation to humans; visual processing

SUMMARY Controlling specific neural circuits across large areas of the brain is a major technology goal of the BRAIN Initiative. To achieve this goal, technologies should ideally provide a combination of spatial, temporal and cell- type specificity and be noninvasive to facilitate their translation across animal models and, ultimately, human patients. Here, we propose an approach to modulating neural circuits noninvasively with spatial, cell-type and temporal specificity. This approach, which we have named Acoustically Targeted Chemogenetics, or ATAC, uses transient focused ultrasound (FUS) blood brain barrier opening (BBBO) to transduce neurons at specific locations in the brain with virally-encoded engineered receptors, which subsequently respond to systemically administered bio-inert compounds to activate or inhibit the activity of these neurons. This technology allows a brief, noninvasive procedure to make one or more specific brain regions capable of being selectively modulated using orally bioavailable compounds. In preliminary experiments, we have implemented this concept in mice by using ATAC to noninvasively target AAV9 viral vectors encoding chemogenetic DREADD receptors to excitatory neurons in the hippocampus, and showing that this enables pharmacological inhibition of memory formation. Building on this proof of concept, we will now scale ATAC to work in non-human primates. This goal is particularly important given the relatively limited success of existing technologies, including optogenetics and conventional chemogenetics, in robust behavioral neuromodulation in larger animals. Scaling ATAC to larger animals requires several innovations beyond the core concept, including evolving viral vectors for more efficient and intersectional transfection of neurons with FUS-BBBO, developing ultrasound methods to overcome skull aberrations and enable precise targeting in large animals, establishing ways of confirming the functionality of ATAC non- invasively with functional imaging, and optimizing the selection and pharmacological administration of chemogenetic ligands for large-animal behavioral studies. In this project, we will first establish the basic capabilities of ATAC in NHPs and integrate them with non-invasive functional imaging, setting a baseline for ATAC performance. Then, we will use a pioneering technology for in vivo evolution of viral vectors to develop AAV viruses specifically optimized to efficiently deliver chemogenetic receptors to brain regions targeted with FUS-BBBO. In parallel, we will develop non-clinical image guidance and aberration correction methods to enable precise targeting and verification of FUS-BBBO in NHPs. This will make it possible for academic groups without access to expensive clinical FUS systems to perform ATAC in larger organisms. Finally, as motivating example applications, we will demonstrate that the optimized ATAC paradigm can be used to inhibit multiple distinct brain regions in macaques, reversibly and repeatably modulating their ability to recognize faces and also apply it in a sensorimotor circuit to alter functional connectivity. We will also show its stability, reliability and non-toxicity.

概括 控制大脑大区域的特定神经回路是大脑的主要技术目标 倡议。为了实现这一目标，理想情况下应提供空间，时间和细胞的组合 键入特异性，无创，以促进它们在动物模型中的翻译，最终人类 患者。在这里，我们提出了一种通过空间，细胞类型和 时间特异性。这种方法，我们将其命名为拟声靶向的化学遗传学或ATAC， 使用瞬态聚焦超声（FUS）血脑屏障开口（BBBO）在特定的 用病毒编码的工程受体在大脑中的位置，随后对系统做出反应 施用的生物启动化合物激活或抑制这些神经元的活性。这项技术允许 简短的无创方法，使一个或多个特定的大脑区域能够选择性调节 使用口服生物利用化合物。在初步实验中，我们通过 使用ATAC非侵入性靶向编码化学遗传DREADD受体的AAV9病毒载体 海马中的神经元，并表明这可以对记忆形成的药理抑制作用。 在此概念证明的基础上，我们现在将扩展ATAC在非人类灵长类动物中工作。这个目标特别是 考虑到现有技术的成功相对有限，包括光遗传学和常规技术的成功很重要 化学遗传学，在较大动物的鲁棒行为神经调节中。将ATAC缩放到大型动物需要 核心概念以外的几项创新，包括发展病毒向量，以提高高效和交叉 通过FUS-BBBO转染神经元，开发了克服颅骨畸变和 在大型动物中启用精确的靶向，建立确认ATAC非 - 的功能的方法 具有功能成像的侵入性，并优化 用于大动画行为研究的化学生成配体。在这个项目中，我们将首先建立基本 ATAC在NHP中的功能并将其与非侵入性功能成像集成在一起，为 ATAC性能。然后，我们将使用开创性的技术进行病毒媒介的体内演变 专门优化的AAV病毒可有效地将化学发生受体传递给针对的大脑区域 FUS-BBBO。同时，我们将开发非临床图像指导和畸变校正方法以实现 NHP中FUS-BBBO的精确靶向和验证。这将使没有 访问昂贵的临床FUS系统以在较大的生物体中执行ATAC。最后，作为激励人心的榜样 应用程序，我们将证明优化的ATAC范式可用于抑制多个不同的大脑 猕猴中的区域，可逆，重复地调节其识别面的能力，并将其应用于 感觉运动电路以改变功能连通性。我们还将显示其稳定性，可靠性和无毒性。