Therapeutic potential for modulation of olfactory basal stem cells

项目摘要/摘要 据估计，至少有12%的人口受到嗅觉丧失的影响。嗅觉丧失，嗅觉丧失 功能障碍可能是由于衰老、先前的感染或损伤，目前还没有有效的治疗方法。虽然 病因可能有所不同，有证据表明，神经源性疲惫，或未能更换或维持 嗅神经元群是许多嗅觉障碍的基础。成人嗅神经发生的调控 因此，这是一个活跃的研究领域。建立在最近的成功基础上，允许净化和培养 成体嗅基干或祖细胞(球状基底层细胞)的鉴定 在嗅觉细胞亚群中的复合体，这一建议询问Polycomb复合体是否以及如何介导 表观遗传修饰影响嗅觉上皮的神经发生和上皮内环境的稳定。 多梳复合体是发育过程中必不可少的表观遗传调节因子，但它们在嗅觉中的作用 维护和更新还没有调查过。使用培养模型，染色体 免疫沉淀-DNA测序(CHIP-SEQ)以及体内方法，Aim1将检验这一假说 多梳复合体调节嗅觉上皮的更新和分化。除了定义 基底细胞的表观遗传调控，AIM2将测试基底干细胞用于修复嗅觉的能力 损坏。这一目标将解决一个主要的翻译问题：基于细胞的疗法能否治疗感觉神经 嗅觉障碍？因为再生的宿主神经元干扰了对术后功能恢复的评估 在小鼠的嗅觉损伤中，我们将采用一种新的可诱导的无嗅觉小鼠模型，在该模型中， 神经元缺乏纤毛，用于测试移植细胞的治疗潜力。这些实验将结合组织学， 电生理学和行为学方法综合评价基底细胞植入 诱导性嗅觉缺失症宿主。此外，该模型还将用于直接测试改变Polycomb的效果 在供体细胞中表达。这些研究在缺乏当前治疗方法的重要临床问题的指导下，将 使用创新的多管齐下的方法来定义以前未曾探索过的嗅觉机械控制 更新和分化，并将为设计基于细胞的感觉神经治疗提供必要的数据 嗅觉丧失继发于损伤或神经性疲惫，如老年性嗅觉障碍。

PROJECT SUMMARY/ABSTRACT It is estimated that at least 12% of the population is affected by olfactory loss. Anosmia, the loss of olfactory function, can be due to aging, prior infection, or injury, and there are currently no effective treatments. Although etiologies may vary, evidence suggests that neurogenic exhaustion, or a failure to replace or maintain the olfactory neuron population, underlies many olfactory disorders. The regulation of adult olfactory neurogenesis is, therefore, an area of active research. Building upon recent successes permitting the purification and culture of adult olfactory basal stem or progenitor cells (globose basal cells), and the identification of Polycomb complexes in subsets of olfactory cells, this proposal asks whether and how Polycomb complex-mediated epigenetic modifications influence neurogenesis and epithelial homeostasis in the olfactory epithelium. Polycomb complexes are essential epigenetic regulators during development, but their roles in olfactory maintenance and renewal have not been investigated. Using a culture model, chromosome immunoprecipitation- DNA sequencing (ChIP-seq), as well as in vivo approaches, Aim1 will test the hypothesis that Polycomb complexes regulate renewal and differentiation in the olfactory epithelium. In addition to defining epigenetic regulation in basal cells, Aim2 will test the ability of basal stem cells to be used to repair olfactory damage. This Aim will address a major translational question: can a cell-based therapy treat a sensorineural anosmia? Because regenerating host neurons interfere with the assessment of functional recovery following olfactory lesions in mice, we will employ a novel inducible anosmia mouse model, in which regenerating neurons lack cilia, for testing therapeutic potential of engrafted cells. The experiments will combine histology, electrophysiology and behavioral approaches to comprehensively evaluate basal cell engraftment into inducible anosmia hosts. Also, this model will be used to directly test the effects of altering Polycomb expression in donor cells. These studies, guided by important clinical problems lacking current treatments, will use innovative multi-pronged approaches to define previously unexplored mechanistic controls of olfactory renewal and differentiation, and will provide essential data to design cell-based therapy for sensorineural olfactory losses secondary to damage or neurogenic exhaustion, such as presbyosmia.