Mentoring Patient Oriented Research in sensory disorders

摘要： 感觉毛细胞是平衡功能所必需的。前庭毛细胞变性导致平衡 功能障碍主要表现为头晕、眩晕。在小鼠身上，有限程度的自发再生 发生在椭圆体中，这是一个前庭器官，检测线加速。此外，还增加了 转录因子Atoh1强健促进小鼠椭圆囊中毛细胞的再生，但 与天然毛细胞相比，再生毛细胞仅部分成熟。在初步实验中，我们 已经鉴定了一种新的AAV-Atoh1结构，该结构可以在 鼠标椭圆体。在再生人类毛细胞以恢复平衡功能的长期目标下， 仍然存在两个主要障碍：1)我们对分子签名的理解不完整 人类毛细胞、支持细胞和毛细胞前体，2)成人内耳组织 并不容易用来测试在动物模型中发现的有前途的疗法。 为了克服这些障碍，我们设计了一种外科方法，从 通常听力和前庭功能正常的已故器官捐赠者。我们已经开始了 收集他们的医疗记录、单细胞转录本和椭圆体的组织切片。在……里面 同时，我们从接受手术切除的前庭神经鞘瘤患者身上获取了椭圆囊。 在这里，我们建议增加器官捐赠者和前庭神经鞘瘤患者的招募。 并描述和验证毛细胞、支持细胞和毛细胞前体在 这两个队列中的成人椭圆体(目标1)。此外，我们将决定我们的小说是否 AAV-Atoh1促进培养的人卵圆囊的毛发再生和成熟(目标2)。 这个奖项的另一个目标是进一步发展我作为一名研究人员和患者导师的职业生涯- 以研究为导向。为了在我之前的板凳研究和指导经验的基础上，这个奖项 旨在保护我的时间，帮助我获得研究人类毛细胞的知识和技能 再生和2)指导其他从事以患者为中心的研究的人。 总而言之，我们将应用最先进的技术(单细胞RNA测序、基因 治疗，生物信息学策略)来研究活体人卵圆囊内前庭毛细胞的再生。我们 已经组建了一支以患者为中心的研究合作者和经验丰富的导师团队。在 在这项为期5年的计划结束时，我们将有1)描绘的人类毛细胞转录，支持 细胞和毛细胞前体，2)揭示了新的AAV-Atoh1是否促进再生和 人类前庭毛细胞的成熟，以及3)增强了我在 以病人为中心的感觉障碍研究。

Abstract: Sensory hair cells are required for balance function. Vestibular hair cell degeneration causes balance dysfunction manifested as dizziness and vertigo. In mice, a limited degree of spontaneous regeneration occurs in the utricle, a vestibular organ detecting linear acceleration. Moreover, addition of the transcription factor Atoh1 robust enhances regeneration of hair cells in the mouse utricle, but regenerated hair cells mature only partially relative to native hair cells. In preliminary experiments, we have characterized a novel AAV-ATOH1 construct leading to regeneration of more mature hair cells in the mouse utricle. With the long-term goal of regenerating human hair cells to restore balance function, two major obstacles remain: 1) we have an incomplete understanding of the molecular signatures of human hair cells, supporting cells and hair cell precursors, 2) adult human inner ear tissues are not readily available to test promising therapeutics discovered in animal models. To overcome these obstacles, we have designed a surgical method to procure live utricles from deceased organ donors who typically have normal auditory and vestibular function. We have begun assembling their medical records, single-cell transcriptomes, and histologic sections of utricles. In parallel, we have procured utricles from vestibular schwannoma patients undergoing surgical resection. Here, we propose to increase the recruitment of organ donors and vestibular schwannoma patients and delineate and validate the transcriptomes of hair cells, supporting cells, and hair cell precursors in adult human utricles in these two cohorts (Aim 1). Furthermore, we will determine whether our novel AAV-ATOH1 enhances hair regeneration and maturation in cultured human utricles (Aim 2). Another goal of this award is to further my career as an investigator and mentor in patient- oriented research. To build on my previous experience in bench research and mentoring, this award is designed to protect my time and help me gain knowledge and skills to 1) study human hair cell regeneration and 2) mentor others pursuing patient-oriented research. In summary, we will apply state-of-the art technologies (single-cell RNA-sequencing, gene therapy, bioinformatic strategies) to study vestibular hair cell regeneration in live human utricles. We have assembled a team of collaborators and experienced mentors in patient-oriented research. At the end of this 5-year proposal, we will have 1) delineated transcriptomes of human hair cells, supporting cells, and hair cell precursors, 2) revealed whether novel AAV-ATOH1 enhances regeneration and maturation of human vestibular hair cells, and 3) enhanced my ability to perform and mentor others in patient-oriented research in sensory disorders.