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。我们之前的应用重点是使用干毛细胞再生 基于细胞的表观遗传学方法。该更新提案的主要目标是确定分子 在正常发育过程中对小鼠毛细胞突触形成至关重要的机制并发育出新的干细胞 基于细胞的方法来重新支配小鼠听觉毛细胞。毛细胞再生获得显着 进步。与毛细胞再生相比，螺旋神经节神经元（SGN）再生已被报道，但 结果相对有限。我们已经确定了一种引导小鼠胚胎干细胞 (ESC) 的逐步方法 分化为具有 SGN 特征的 ESC 衍生的 SGN 样神经元 (ESN)。截至目前，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。然而，无论 这些神经联系是否是真正的 SGN-毛细胞突触仍不清楚。这些神经元的功能 尚未在突触水平上确定接触。再生关键的分子机制 功能性重新支配毛细胞的神经元仍然不清楚。因此，迫切需要使用ESN来 重新支配毛细胞并确定重新神经支配的关键分子机制。 EphrinB家族蛋白 与 EphB 受体（跨膜受体酪氨酸激酶家族）结合。 EphrinB 信号传导参与 在各种发育程序中，包括突触发生。先前的研究表明 EphrinB1 信号传导对于兴奋性突触诱导至关重要。我们的初步数据表明 EphrinB 信号传导的作用 ESN-毛细胞神经支配。根据之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方面发挥关键作用。到 检验这一假设，提出了三个互补的具体目标。目标 1 将确定神经元的作用 Efnb1 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 ESN-毛细胞神经支配的研究。目标 1 和 2 的结果将指导毛细胞突触再生的设计 研究。在目标 3 中，我们将确定 EphrinB1 调节 ESN-毛细胞神经支配的程度。 该提案的完成将确定干细胞衍生神经元关键的分子机制 重新支配感觉毛细胞。确定 Efnb1 在毛细胞神经支配中的作用将指导突触 再生研究，将转化为治疗听力损失患者的临床试验。因此， 这项工作的成果将为探索基于干细胞的多学科方法开辟新途径 再生毛细胞突触并恢复退伍军人和平民患者的听力功能。