Cellular therapy for the inner ear

内耳细胞疗法

• 批准号: 7258349
• 负责人: Albert Edge
• 金额: $ 28.64万
• 依托单位: MASSACHUSETTS EYE AND EAR INFIRMARY
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-08-12 至 2008-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7258349
• 关键词: Acute; Address; Adult; Afferent Neurons; Age; Animal Model; Biological Assay; Biological Models; Brain; Bungarotoxins; Candidate Disease Gene; Cell Differentiation process; Cell Line; Cell Therapy; Cells; Characteristics; Chromosome Pairing; Cochlea; Cochlear nucleus; Condition; Coupled; Development; Ear; Engraftment; Environment; Etiology; Family; Gene Expression; Gene Silencing; Genetic; Gerbils; Goals; Growth Factor; Growth Factor Overexpression; Hair Cells; Histologic; Immunohistochemistry; In Vitro; Labyrinth; Ligands; Luciferases; Measures; Mus; Natural regeneration; Nature; Nerve; Nervous system structure; Neurodegenerative Disorders; Neurons; Neuropilin-1; Neuropilins; Numbers; Organ of Corti; Ouabain; Parkinson Disease; Pathway interactions; Process; Recovery of Function; Reporter; Role; Sensorineural Hearing Loss; Sensory; Series; Signal Pathway; Small Interfering RNA; Source; Spinal Ganglia; Staging; Stem cells; Synapses; System; Testing; Therapeutic; Time; Tissues; Transplantation; Undifferentiated; Withdrawal; Work; axonal guidance; cell type; design; desire; embryonic stem cell; fetal; ganglion cell; in vitro Model; in vivo; in vivo Model; inner ear diseases; nerve stem cell; nerve supply; neuron loss; neuronal guidance; progenitor; promoter; receptor; reconstitution; reinnervation; relating to nervous system; research study; response; sound; spiral ganglion; success; synaptogenesis; transcription factor

DESCRIPTION (provided by applicant): Loss of cochlear neurons and hair cells is a major cause of sensorineural hearing loss. One approach to potential treatments is the transplantation of exogenous progenitor cells. Our strategy for developing cell based therapies for inner ear disorders is to study the reinnervation of a de-afferented organ of Corti. We take this approach because 1) only a single cell type, the spiral ganglion cell, must be replaced to see functional recovery; 2) partial regeneration of damaged tissue by cell replacement has been successful in animal models of other neurodegenerative diseases, such as Parkinson's, suggesting that neuronal circuits in the adult nervous system can be reconstituted if appropriate progenitor cells are introduced and 3) understanding the mechanisms underlying re-formation of neural connections to hair cells in the adult ear is important to any therapeutic approach to sensorineural hearing loss. The Specific Aims comprise four inter-related experiments to probe key variables likely to influence the success of hair cell reinnervation by transplanted neural progenitor cells. In Aim 1 the developmental expression by spiral ganglion neurons of several families of neuronal guidance molecules will be studied by immunohistochemistry. In Aim 2 we use de-afferented cochlear explants to study the reinnervation of hair cells by grafted progenitors, in vitro, by modifying characteristics of the donor cells (e.g. silencing expression of selected guidance molecules or controlling degree of differentiation) or the host environment (e.g. developmental age of the explant or etiology of the initial de-afferentation). In Aim 3 we build on the in vitro results to choose donor cells for in vivo transplantation into a chemically de-afferented gerbil ear, in which the sensory cells are intact but spiral ganglion cells are destroyed. In this model system, success of reinnervation can be assayed both functionally and histologically. In Aim 4 we use a reporter assay to determine conditions needed to bias differentiation of stem cells to a lineage corresponding to sensory neurons by adding specific growth factors in a timed sequence, and by over expressing transcription factors in the pathway to sensory neurons. Clonal cell lines isolated under this Aim will serve as additional donor cell types to be studied in Aims 3 and 4.

描述（由申请人提供）：耳蜗神经元和毛细胞的丧失是感音神经性听力损失的主要原因。一种潜在治疗方法是移植外源祖细胞。我们开发基于细胞的内耳疾病疗法的策略是研究传入神经缺失的柯蒂氏器官的神经支配。我们采用这种方法是因为：1）只有一种细胞类型，即螺旋神经节细胞，必须被替换才能看到功能恢复； 2) 在其他神经退行性疾病（例如帕金森病）的动物模型中，通过细胞替代实现受损组织的部分再生已取得成功，这表明如果引入适当的祖细胞，可以重建成人神经系统中的神经元回路；3）了解成人耳朵中毛细胞神经连接重建的机制对于感音神经性听力损失的任何治疗方法都很重要。具体目标包括四个相互关联的实验，以探讨可能影响移植神经祖细胞重新神经支配毛细胞成功的关键变量。在目标 1 中，将通过免疫组织化学研究几个神经元引导分子家族的螺旋神经节神经元的发育表达。在目标 2 中，我们使用去传入耳蜗外植体，通过修改供体细胞的特征（例如，沉默选定引导分子的表达或控制分化程度）或宿主环境（例如，外植体的发育年龄或初始传入去传入的病因），在体外研究移植祖细胞对毛细胞的神经支配。在目标 3 中，我们在体外结果的基础上选择供体细胞进行体内移植到化学去传入沙鼠耳朵中，其中感觉细胞完整，但螺旋神经节细胞被破坏。在这个模型系统中，神经支配的成功可以从功能上和组织学上进行测定。在目标 4 中，我们使用报告分析来确定将干细胞分化为与感觉神经元相对应的谱系所需的条件，方法是按时间顺序添加特定的生长因子，并在通往感觉神经元的途径中过度表达转录因子。在此目标下分离的克隆细胞系将作为目标 3 和 4 中研究的其他供体细胞类型。