Regeneration of auditory synaptic contacts using stem cell based approaches

使用基于干细胞的方法再生听觉突触接触

• 批准号: 9263690
• 负责人: Zhengqing Hu
• 金额: $ 32.3万
• 依托单位: WAYNE STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-05-01 至 2019-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9263690
• 关键词: Address; Adult; Affinity; Animals; Astrocytes; Auditory; Auditory Evoked Potentials; Auditory system; Binding; Biological; Brain Stem; Cell Nucleus; Cells; Chemicals; Chronic; Cochlea; Cochlear nucleus; Coculture Techniques; Conditioned Culture Media; Cues; Data; Disease; Dose; Ensure; Future; Gene Expression; Generations; Glutamates; Goals; Hair Cells; Hearing; Human; Impairment; Implant; In Vitro; Knockout Mice; Labyrinth; Lead; Ligands; Measures; Mediating; Modeling; Mus; Natural regeneration; Neurites; Neurons; Ouabain; Pilot Projects; Play; Proteins; RNA Interference; Rattus; Research; Retinal Ganglion Cells; Role; Stem cells; Supplementation; Synapses; Testing; Thrombospondin 1; Vestibular ganglion; auditory pathway; base; deafness; gabapentin; hearing impairment; implantation; improved; in vivo; in vivo Model; in vivo regeneration; injured; inner ear diseases; knock-down; nerve stem cell; neurotrophic factor; neurotropin; overexpression; postnatal; public health relevance; receptor; restoration; sensory system; spiral ganglion; success; synaptogenesis

DESCRIPTION (provided by applicant): Spiral ganglion neurons (SGNs) and their synapses with the cochlear nucleus (CN) are important components of the auditory system that are impaired in a variety of auditory disorders. Currently, no biological approach exists to regenerate damaged SGNs. Our long-term aims of this proposal are to use stem cell (SC)-derived neurons to replace injured SGNs and regenerate afferent CN synapses that connect the cochlea to the brainstem. In this proposal, we will focus on afferent synapse generation between SC-derived glutamatergic neurons and CN neurons using in vitro and in vivo models. Regenerating the connections between hair cells and SC-derived neurons will be examined in a separate project. A recent study shows that implanted SC-derived cells improved the evoked auditory responses of deafened animals. However, the mechanism (protein-receptor interaction) whereby synapse formation occurs between implanted cells and host CN neurons remains unknown. We have identified neural SCs from mouse cochlear-vestibular ganglia and induced these SCs to differentiate into glutamatergic neurons (ScNs). We observed that neurotrophins stimulated neurite extensions from ScNs. However, neurotrophin supplementation did not significantly promote synaptogenesis between ScNs and CN neurons. Therefore, identification of mechanisms that stimulate auditory synapse regeneration is a major challenge to the restoration of auditory function. Recently, we developed a co-culture model using ScNs and mouse CN neurons to address this issue. We hypothesize that astrocyte-released thrombospondin-1 (TSP1) and the alpha2delta-1 (�2�-1) receptor are critical for SC-based synaptogenesis. To test this hypothesis, we propose the following specific aims: Aim 1: Determine whether TSP1 is a critical stimulator of ACM-induced synaptogenesis in vitro; Aim 2: Investigate the role of �2�1 in ACM-induced synaptogenesis in co-cultures; Aim 3: Examine the roles of TSP1 and �2�1 in CN synapse regeneration in vivo. This proposal studies a protein and a receptor that appear to be critical for the induction of synaptogenesis between SC-derived cells and CN neurons. When our in vitro and in vivo models are optimized in this proposal, we will study the strategies to develop and refine synaptic connections to ensure that the circuit is properly wired in a tonotopic organization in our future research. Identification of proteins and receptors that are important fo CN synapse formation not only will be fundamental to auditory pathway regeneration, but also will provide cues for synapse regeneration in other sensory systems.

描述（由应用提供）：螺旋神经节神经元（SGN）及其与耳蜗核（CN）的突触是听觉系统的重要组成部分，在多种听觉障碍中受到损害。目前，没有生物学方法可以再生 损坏的SGNS。该提案的长期目的是使用干细胞（SC）衍生的神经元来替代受伤的SGN和再生传入的CN突触，将耳蜗连接到脑干。在此提案中，我们将专注于使用体外和体内模型之间SC衍生的谷氨酸能神经元和CN神经元之间的传入突触产生。将在一个单独的项目中检查毛细胞与SC衍生神经元之间的连接。最近的一项研究表明，植入的SC衍生细胞改善了被破坏动物的诱发听觉反应。但是，在植入细胞和宿主CN神经元之间发生突触形成的机制（蛋白质受体相互作用）仍然未知。我们已经鉴定出小鼠人工耳蜗卵巢神经节的神经元SC，并诱导这些SC分化为谷氨酸能神经元（SCN）。我们观察到神经营养蛋白刺激了SCN的神经蛋白延伸。然而，神经营养蛋白补充并未显着促进SCN和CN神经元之间的突触发生。因此，识别刺激听觉突触再生的机制是对听觉功能恢复的主要挑战。最近，我们使用SCN和Mouse CN神经元开发了共同培养模型来解决此问题。我们假设星形胶质细胞释放的血小板传播-1（TSP1）和α2Delta-1（�2。1）受体对于基于SC的基于SC的突触发生至关重要。为了检验这一假设，我们提出了以下特定目的：目标1：确定TSP1是否是ACM诱导的突触发生的关键刺激剂； AIM 2：研究�2。1在ACM诱导的共培养中的突触发生中的作用； AIM 3：检查TSP1和�2.1在体内突触再生中的作用。该建议研究了一种蛋白质和接收器，这对于SC衍生细胞和CN神经元之间的突触发生至关重要。当我们的体外和体内模型在此提案中优化时，我们将研究开发和完善突触连接的策略，以确保电路正确连接到吨位中 在我们未来的研究中组织。鉴定重要的FO CN突触形成的蛋白质和接收器不仅对听觉途径再生至关重要，而且还将为其他感觉系统中的突触再生提供线索。