Transparent neural interface for in vivo interrogation of human organoids

用于体内询问人体类器官的透明神经接口

• 批准号: 10204516
• 负责人: Anna Devor
• 金额: $ 27.03万
• 依托单位: BOSTON UNIVERSITY (CHARLES RIVER CAMPUS)
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-08-01 至 2021-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10204516
• 关键词: Transparent; neural; interface; vivo; interrogation

ABSTRACT Recent advances in pluripotent stem cell technology have enabled generation of neuronal cell lines and cerebral organoids from human embryonic stem cells (hESCs) as well as human induced pluripotent stem cells (hiPSCs) derived from peripheral tissues. These organoids are self-assembled, 3D cellular structures that resemble early developmental stages of the human brain opening unprecedented opportunities for investigation of human neuronal network-level dysfunction underlying developmental brain disease. However, the lack of the natural brain microenvironment in cultured organoids can influence the phenotype and maturation of the reprogrammed neurons. To mitigate this limitation, we recently transplanted human cerebral organoids into the mouse brain and demonstrated their differentiation and vascularization using 2- photon imaging through cranial “windows” made of glass. In the proposed study, we will replace these windows with optically transparent graphene electrode microgrids, developed by members of our team, to enable multimodal longitudinal monitoring and interrogation of neuronal activity in the graft and the surrounding host neuronal circuits. The in vivo organoid transplantation and graphene electrode arrays are existing technologies providing Scientific Premise to the proposed project – without these parts in place, we would be building a bridge too far. Our current goal is to combine these technologies creating a synergistic and transformative result. Critically, this combination will allow examination of cell-type specific spiking (detected with 2-photon imaging) referenced to large-scale network events arising either from the organoid or the host cortex (detected as Local Field Potentials, LFPs, by a graphene electrode grid). We know from our prior work that organoids achieve sufficient laminar organization and synaptic connectivity to generate LFPs, adding to the Scientific Premise. We will engineer implantable graphene devices that would adhere to the cortical surface and the organoid to enable stable, longitudinal recordings, imaging, and photostimulation in mice transplanted with human cortical organoids (Aim 1). Then, we will provide a proof-of-principle demonstration of the unique advantage of this multimodal technology for studying the evolution of organoid activity during its maturation in vivo. To this end, we will focus on participation of specific cell types in LFP events originating from either the organoid or the host (Aim 2). All experiments will be performed in awake mice without confounds of anesthesia. Since transplantation of human cerebral organoids in the mouse brain is still in infancy, this project will deliver a much needed tool for comprehensive functional assessment of this novel biological model system. Further along the road (outside the current scope), this model system will find its use for investigating aspects of human brain development and developmental disorders.

抽象的 多能干细胞技术的最新进展已经能够产生神经细胞系和 来自人类胚胎干细胞（hESC）以及人类诱导多能干细胞的大脑类器官 （hiPSC）源自外周组织，这些类器官是自组装的 3D 细胞结构。 类似于人类大脑的早期发育阶段，为人类大脑提供了前所未有的机会 研究发育性脑疾病背后的人类神经网络水平功能障碍。 培养的类器官中缺乏自然的大脑微环境会影响表型和 为了减轻这种限制，我们最近移植了人类。 将脑类器官植入小鼠大脑并使用 2- 展示其分化和血管化 在拟议的研究中，我们将通过玻璃制成的颅“窗”进行光子成像。 由我们团队成员开发的带有光学透明石墨烯电极微网格的窗户， 能够对移植物和移植物中的神经活动进行多模式纵向监测和询问 周围宿主神经回路。 体内类器官移植和石墨烯电极阵列正在提供现有技术 拟议项目的科学前提——如果没有这些部件，我们也将建造一座桥梁 我们当前的目标是将这些技术结合起来，创造出协同和变革的结果。 至关重要的是，这种组合将允许检查细胞类型特异性尖峰（通过 2 光子成像检测） 涉及由类器官或宿主皮层（检测为本地皮层）引起的大规模网络事件 场电位，LFP，通过石墨烯电极网格）我们从之前的工作中知道类器官可以实现。 足够的层状组织和突触连接来生成 LFP，增加了科学前提。 我们将设计可植入的石墨烯设备，该设备将粘附在皮质表面并 该类器官能够在小鼠中实现稳定的纵向记录、成像和光刺激 然后，我们将提供原理验证。 展示了这种多模态技术在研究进化过程中的独特优势 为此，我们将重点关注特定类器官在体内成熟过程中的参与。 LFP 事件中源自类器官或宿主的细胞类型（目标 2）。 在清醒的小鼠中进行，没有麻醉的干扰。 由于将人类大脑类器官移植到小鼠大脑中仍处于起步阶段，该项目将实现 这是对这种新型生物模型系统进行全面功能评估所急需的工具。 沿着道路（当前范围之外），该模型系统将用于调查以下方面： 人类大脑发育和发育障碍。

项目成果

Chronic Cranial Windows for Long Term Multimodal Neurovascular Imaging in Mice.

用于小鼠长期多模式神经血管成像的慢性颅窗。

• 发表时间: 2020
• 作者: Kılıç, Kıvılcım;Desjardins, Michèle;Tang, Jianbo;Thunemann, Martin;Sunil, Smrithi;Erdener, Şefik Evren;Postnov, Dmitry D;Boas, David A;Devor, Anna
• 通讯作者: Devor, Anna