Developing novel stem cell-based approaches to treat hearing loss

开发基于干细胞的新型方法来治疗听力损失

• 批准号: 10641152
• 负责人: Zhengqing Hu
• 金额: --
• 依托单位: JOHN D DINGELL VA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-12-01 至 2027-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10641152
• 关键词: Acoustics; Affect; American; Animal Model; Auditory; Binding Proteins; Cell Culture Techniques; Cells; Clinical Trials; Cochlea; Coculture Techniques; Data; Denervation; Development; Ephrin B Receptor; Ephrin Receptor EphB1; Ephrins; Epigenetic Process; Family; Future; Genes; Goals; Hair Cells; Hearing; Hearing problem; In Vitro; Knockout Mice; Knowledge; Labyrinth; MYO7A gene; Membrane; Methods; Modeling; Molecular; Mus; Natural regeneration; Neurons; Neurophysiology - biologic function; Outcome; Patients; Pilot Projects; Play; Protein Family; Proteins; Publishing; Receptor Protein-Tyrosine Kinases; Regenerative research; Reporting; Role; Sensory Hair; Signal Transduction; Source; Synapses; Testing; Translating; Translational Research; Transplantation; Veterans; Work; conditional knockout; design; embryonic stem cell; functional restoration; hair cell regeneration; hearing impairment; hearing restoration; in vivo; insight; interdisciplinary approach; myosin VI; nerve stem cell; neural; neuron regeneration; novel; postnatal; programs; protein expression; regeneration function; reinnervation; ribbon synapse; self-renewal; sound; spiral ganglion; stem cell based approach; stem cells; synaptogenesis

The long-term aim of this project is to develop a stem cell-based approach to regenerate the function of damaged auditory cells and synaptic connections. Our previous application focused on hair cell regeneration using stem cell-based epigenetic approaches. The major objectives of this renewal proposal are to determine the molecular mechanism critical for mouse hair cell synapse formation during normal development and develop a novel stem cell-based approach to reinnervate mouse auditory hair cells. Hair cell regeneration has obtained significant progress. Compared to hair cell regeneration, spiral ganglion neuron (SGN) regeneration has been reported but with relatively limited results. We have identified a stepwise method to guide mouse embryonic stem cells (ESCs) to differentiate into ESC-derived SGN-like neurons (ESNs) that showed SGN features. Up to date, only a few reports show that stem cell-derived neurons can form neural contacts with mouse hair cells. However, whether these neural contacts are bona fide SGN-hair cell synapses remains unclear. The function of these neural contacts has not been determined at the synaptic level. The molecular mechanism critical for regenerated neurons to functionally reinnervate hair cells remains obscure. Therefore, there is a critical need to use ESNs to reinnervate hair cells and determine the molecular mechanism critical for reinnervation. EphrinB family proteins bind to the EphB receptors, a family of transmembrane receptor tyrosine kinases. EphrinB signaling is involved in a variety of developmental programs, including synaptogenesis. Previous studies show that EphrinB1 signaling is critical for excitatory synaptic induction. Our preliminary data suggest the role of EphrinB signaling in ESN-hair cell reinnervation. Based on previous and our preliminary data, we hypothesize that EphrinB signaling may play a critical role in regulating neurons to form functional synapses with sensory hair cells. To test this hypothesis, three complementary specific aims are proposed. Aim 1 will determine the role of neuronal Efnb1 in SGN-hair cell synapse formation during development. Aim 2 will determine the molecular mechanism of ESN-hair cell reinnervation. Results of Aims 1 and 2 will guide the design of hair cell synapse regeneration research. In Aim 3, we will determine the extent to which EphrinB1 regulates ESN-hair cell reinnervation. Completion of this proposal will determine the molecular mechanisms critical for stem cell-derived neurons to reinnervate sensory hair cells. Identifying the role of Efnb1 in hair cell reinnervation will guide the synapse regeneration research, which will be translated into clinical trials to treat hearing loss patients. Therefore, the outcomes of this work will open new avenues to explore a stem cell-based multidisciplinary approach to regenerate hair cell synapses and restore the hearing function of Veteran and civilian patients.

该项目的长期目标是开发一种基于干细胞的方法来再生受损的功能。 听觉细胞和突触连接。我们以前的应用程序集中在毛细胞再生使用干 基于细胞的表观遗传学方法。本更新提案的主要目标是确定 在正常发育过程中小鼠毛细胞突触形成的关键机制，并开发一种新的干细胞 基于细胞的方法来重新神经支配小鼠听觉毛细胞。毛细胞再生已获得显着 中求进工作总与毛细胞再生相比，已经报道了螺旋神经节神经元（SGN）再生，但 结果相对有限。我们已经确定了一个逐步的方法来引导小鼠胚胎干细胞（ESCs） 以分化成显示SGN特征的ESC衍生的SGN样神经元（ESN）。到目前为止，只有少数 报告显示干细胞衍生的神经元可以与小鼠毛细胞形成神经接触。但无论 这些神经接触是真正的SGN-毛细胞突触仍然不清楚。这些神经元的功能 接触尚未在突触水平上确定。再生的关键分子机制 神经元在功能上重新支配毛细胞仍然不清楚。因此，迫切需要使用ESN来 神经再生毛细胞和确定神经再生的关键分子机制。EphrinB家族蛋白 结合EphB受体，一个跨膜受体酪氨酸激酶家族。EphrinB信号通路参与 包括突触发生在内的多种发育过程中。先前的研究表明，EphrinB 1 信号传导对于兴奋性突触诱导是至关重要的。我们的初步数据表明EphrinB信号传导的作用 在ESN-毛细胞神经再生中。基于先前和我们的初步数据，我们假设EphrinB 信号传导可能在调节神经元与感觉毛细胞形成功能性突触中起关键作用。到 为了验证这一假设，提出了三个互补的具体目标。目的1将确定神经元的作用 Efnb 1在发育过程中SGN-毛细胞突触形成中的作用目标2将确定分子机制 毛细胞神经再生目的1和2的结果将指导毛细胞突触再生的设计 research.在目标3中，我们将确定EphrinB 1调节ESN毛细胞神经再支配的程度。 完成这项提案将确定干细胞衍生的神经元的关键分子机制， 重新支配感觉毛细胞。确定Efnb 1在毛细胞神经再支配中的作用将指导突触 再生研究，这将转化为临床试验，以治疗听力损失患者。因此 这项工作的成果将开辟新的途径，探索基于干细胞的多学科方法， 再生毛细胞突触，恢复退伍军人和平民患者的听力功能。