Sonic Hedgehog Signaling in Inner Ear Organoid Development

内耳类器官发育中的 Sonic Hedgehog 信号传导

• 批准号: 9336149
• 负责人: Emma Longworth-Mills
• 金额: $ 2.83万
• 依托单位: INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-01 至 2019-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9336149
• 关键词: Aging; Agonist; Area; Auditory; Biochemical; Biological Models; Cell Therapy; Cell model; Cells; Cellular biology; Cochlea; DNA Sequence Alteration; Derivation procedure; Development; Dimensions; Disease; Dorsal; ES Cell Line; Electrophysiology (science); Exhibits; FBXO2 gene; Foundations; Functional disorder; Generations; Gravity Perception; Hair Cells; Hearing; Human; Immunohistochemistry; In Vitro; Inherited; Laboratories; Labyrinth; Loudness; Methods; Modeling; Molecular Biology; Morphogenesis; Morphology; Mus; Natural regeneration; Nature; Noise; Organ; Organ of Corti; Organoids; Otic Vesicle; Pathology; Pattern; Pharmaceutical Preparations; Play; Pluripotent Stem Cells; Population; Preclinical Drug Evaluation; Process; Property; Quantitative Reverse Transcriptase PCR; Reporter; Research; Role; SHH gene; Sensorineural Hearing Loss; Sensory; Sensory Hair; Sensory Receptors; Signal Transduction; Sonic Hedgehog Pathway; Stem cells; Structure; Study models; System; Testing; Tissues; Type II Hair Cell; Vestibular Hair Cells; cell type; deafness; embryonic stem cell; human pluripotent stem cell; human tissue; in utero; inner ear development; inner ear diseases; novel; ototoxicity; progenitor; purmorphamine; small molecule; smoothened signaling pathway; sound; three dimensional cell culture

ABSTRACT Loss of the finite cochlear hair cells in the inner ear results in sensorineural deafness. Human cochlear hair cells do not regenerate, and there is no cure for deafness. Our lab has recently established a novel three-dimensional culture system for deriving functional sensory hair cells from mouse and human pluripotent stem cells. A major limitation of this approach, however, is that derived hair cells exhibit structural, biochemical and electrophysiological properties of gravity-sensing vestibular hair cells. The processes underlying the commitment to cochlear versus vestibular fate in inner ear sensory hair cell development are poorly understood. Previous studies have shown that establishment of a dorsal-ventral (DV) axis in the developing otic vesicle is necessary for proper morphogenesis of both auditory and vestibular inner ear structures. Sonic hedgehog signaling has been shown to play a key role in precise DV patterning of the otic vesicle. In Specific Aim 1, I will characterize the nature of DV patterning in otic vesicles derived using our three-dimensional inner ear culture model, and determine whether commitment to a vestibular fate is due to a lack of ventralizing signals. In Specific Aim 2, I will assess whether modulation of Sonic hedgehog signaling via small molecule application in our culture model is able to induce differentiation of cochlear cell types. Stem cell-derived cochlear hair cells may serve as a potent human model system to study pathophysiology of various forms of hereditary deafness. Such an advance could profoundly impact our understanding of disease processes that normally occur in utero. Furthermore, an in vitro system recapitulating both cochlear and vestibular sensory cell development is amenable to high-throughput drug screening to identify compounds that either enhance inner ear differentiation, promote the survival of inner ear sensory cells, or have ototoxic effects.

摘要 内耳中有限耳蜗毛细胞的丧失导致感音神经性耳聋。人类 耳蜗毛细胞不能再生，耳聋也无法治愈。我们的实验室最近 建立了一种新的三维培养系统，用于从 小鼠和人类多能干细胞。然而，这种方法的一个主要限制是， 毛细胞具有重力感应的结构、生物化学和电生理特性 前庭毛细胞耳蜗与前庭命运的潜在承诺过程 内耳感觉毛细胞的发育知之甚少。以前的研究已经表明 在发育中的耳泡中建立背腹（DV）轴对于正确的 听觉和前庭内耳结构的形态发生。音速刺猬信号 已显示在耳泡的精确DV图案化中起关键作用。在具体目标1中，我将 表征使用我们的三维内耳导出的耳泡中DV图案的性质 文化模式，并确定是否承诺前庭的命运是由于缺乏腹 信号.在具体目标2中，我将评估是否通过小的 在我们的培养模型中应用分子能够诱导耳蜗细胞类型的分化。干 细胞来源的耳蜗毛细胞可以作为一个有效的人类模型系统，以研究病理生理学 各种遗传性耳聋这样的进步可能会深刻地影响我们对 通常在子宫内发生的疾病过程此外，一个体外系统概括了两者， 耳蜗和前庭感觉细胞的发育适合于高通量药物筛选， 鉴定增强内耳分化、促进内耳存活的化合物 感觉细胞，或具有耳毒性作用。