Ultrasonic modulation of cellular electrical signaling

细胞电信号的超声波调制

基本信息

批准号：
10352016
负责人：
Em Ben Sorum
金额：
$ 11.59万
依托单位：
UNIVERSITY OF CALIFORNIA BERKELEY
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-01-01 至 2023-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10352016
关键词：
Acoustics Action Potentials Address Architecture Area Axon Behavior Biological Biomedical Technology Biophysical Process Biophysics Brain Brain imaging Calcium Calcium Binding Cell Line Cell membrane Cells Chemical Agents Clinical Cultured Cells Cystic Fibrosis Transmembrane Conductance Regulator Data Development Electrophysiology (science)Engineering FDA approved Focused Ultrasound Goals Hybrids Hypersensitivity Image In Vitro Ion Channel Ions Knowledge Laboratory Finding Light Lipids Mechanics Membrane Membrane Lipids Mentors Modality Modeling Modernization Molecular Motion Mus Mutagenesis Mutate Nature Nerve Neurons Neurosciences Permeability Phase Physiological Physiology Potassium Channel Proteins Protocols documentation Radial Ranvier&apos s Nodes Research Resolution Signal Transduction Site Slice Stimulus Structure Surveys System Techniques Testing Time Tissues Training Transgenic Organisms Translating Ultrasonic wave Ultrasonics Validation Work Xenopus oocyte base bone experimental study fluorophore hippocampal pyramidal neuron in vivo mechanical energy minimally invasive neural circuit neuronal excitability neurophysiology neuroregulation novel novel strategies optogenetics programs proteoliposomes reconstitution relating to nervous system response skills sound tool trait ultrasound vibration white matter

项目摘要

Ultrasound (US) neuromodulation (NM) utilizes mechanical energy from sound to modulate the physiology of excitable cells through mechanosensitive ion channels (MSIC). Its uninvasive bone-penetrating nature combined with a unique focusing capability provides advantages over optogenetics and chemogenetics. Recently, the FDA has approved transcranial USNM. There is a lack of a molecular understanding on how US modulates cellular excitability. Furthermore, not all cells express channels that respond to US. To address these issues, novel experimental systems and techniques will be implemented to extract mechanistic biophysical information and engineer sonogenetic tools for robust transgenic expression. The biophysical effects of US on TRAAK K+ channels were recently characterized because of its potential to serve as a sonogenetic silencer of neurons. The endogenous expression of TRAAK at the nodes of Ranvier also makes it an attractive candidate for inhibitory NM when targeting native myelinated axons in the white matter. During the mentored phase (Aim 1, K99), the fundamental biophysical effects of US will continue to be characterized on TRAAK and other channels. This includes experiments to calculate tension and surveying optimal US parameters to maximize channel stimulation. Preliminary data suggests that the CFTR Cl- channel is sensitive to US. Other MSIC will also be screened to identify those sensitive to US. The next aim (Aim 2, K99/R00) looks to optimize TRAAK and other MSIC into sonogenetic tools. To transform TRAAK into a US-hypersensitive action potential generator, structure guided mutagenesis will be used to increase its sensitivity to ultrasound and permeability to Na+. Aim 3 (R00 phase) looks to implement NM by stimulating endogenous MSIC and transgenically expressed sonogenetic tools. They will be activated in vitro in mouse brain slices and cultured neurons, and in vivo in live mice. In summary, this proposal looks to screen for and optimize the activation of endogenous MSIC with US, and also engineer novel sonogenetic tools. In a short period of time, this work will advance our understanding of the effects of US on MSIC and NM. The long-term goal is to implement these tools for clinical use in minimally invasive NM. This research program will generate a platform of complementary techniques and applicable knowledge that can also be applied by others for further studies in sonogenetics and USNM. Dr. Sorum's mentors, Profs. Brohawn and Adesnik, have expertise that spans many areas of neuroscience including mechanobiology, MSIC structure/function, optogenetics, NM, neural circuits, and behavior. Two additional years of training will allow him to fully develop skills in molecular, cellular and systems neuroscience and merge them with USNM and sonogenetics.

超声（US）神经调节（NM）利用声音的机械能通过机械敏感离子通道（MSIC）调节可兴奋细胞的生理机能。其非侵入性骨穿透性质与独特的聚焦能力相结合，提供了优于光遗传学和化学遗传学的优势。最近，FDA批准了经颅USNM。目前缺乏对 US 如何调节细胞兴奋性的分子理解。此外，并非所有细胞都表达对 US 做出反应的通道。为了解决这些问题，将实施新的实验系统和技术来提取机械生物物理信息并设计声遗传学工具以实现稳健的转基因表达。 US 对 TRAAK K+ 通道的生物物理效应最近得到了表征，因为它有可能充当神经元的声遗传学沉默器。当靶向白质中的天然有髓鞘轴突时，TRAAK 在 Ranvier 节点的内源性表达也使其成为抑制性 NM 的有吸引力的候选者。在指导阶段（目标 1、K99），US 的基本生物物理效应将继续在 TRAAK 和其他渠道上得到表征。这包括计算张力和调查最佳超声参数以最大化通道刺激的实验。初步数据表明CFTR Cl-通道对US 敏感。其他 MSIC 也将受到筛查，以识别对美国敏感的人。下一个目标（目标 2、K99/R00）旨在将 TRAAK 和其他 MSIC 优化为声遗传学工具。为了将 TRAAK 转变为美国超敏动作电位发生器，将使用结构引导诱变来增加其对超声波的敏感性和对 Na+ 的渗透性。目标 3（R00 阶段）希望通过刺激内源 MSIC 和转基因表达的声遗传学工具来实施 NM。它们将在体外的小鼠脑切片和培养的神经元中以及在活体小鼠的体内被激活。总之，该提案旨在通过 US 筛选和优化内源 MSIC 的激活，并设计新颖的声遗传学工具。在短时间内，这项工作将增进我们对美国对 MSIC 和 NM 影响的理解。长期目标是将这些工具应用于微创 NM 的临床应用。该研究计划将生成一个补充技术和适用知识的平台，其他人也可以将其应用于声遗传学和 USNM 的进一步研究。 Sorum 博士的导师，教授。 Brohawn 和 Adesnik 拥有跨越神经科学许多领域的专业知识，包括机械生物学、MSIC 结构/功能、光遗传学、NM、神经回路和行为。另外两年的培训将使他能够充分发展分子、细胞和系统神经科学方面的技能，并将其与 USNM 和声遗传学相结合。