Driving the Progeny of Olfactory HBC Stem Cells toward Neuronal Differentiation

驱动嗅觉 HBC 干细胞后代向神经元分化

• 批准号: 10527167
• 负责人: JAMES E. SCHWOB
• 金额: $ 24.75万
• 依托单位: TUFTS UNIVERSITY BOSTON
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-01 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10527167
• 关键词: Address; Adopted; Adult; Afferent Neurons; Age; Aged, 80 and over; Aging; Anosmia; Area; Automobile Driving; Back; Basal Cell; Biological; Biological Assay; Biology; Birth; Cell Culture System; Cell Differentiation process; Cells; Curcumin; Data; Disease; Dominant-Negative Mutation; Environment; Epithelial; Epithelial Cells; Equilibrium; Evaluation; Failure; Formulation; Functional disorder; GDF11 gene; Gene Activation; Gene Expression Profile; General Population; Generations; Genetic Transcription; Goals; Human; In Situ; In Vitro; Joints; Knock-out; Laboratories; Lamina Propria; Lesion; Metaplasia; Minority; Mission; Molecular; Mus; National Institute on Deafness and Other Communication Disorders; Natural regeneration; Neuroglia; Neuronal Differentiation; Neurons; Nose; Nutritional status; Olfactory Epithelial Cell; Olfactory Epithelium; Olfactory dysfunction; Outcome; Pathologic; Pathology; Pathway interactions; Phase; Phorbol Esters; Population; Preclinical Testing; Process; Proteomics; Proto-Oncogene Protein c-kit; Quality of life; Quercetin; Rejuvenation; Reserve Stem Cell; Resveratrol; Rodent; Safety; Sensory; Signal Transduction; Smell Perception; Stromal Cells; Technology; Testing; Therapeutic; Tissues; Transfection; Transplantation; Work; Xenograft procedure; age related; aged; airway epithelium; base; directed differentiation; efficacy testing; efficacy validation; epithelial injury; epithelial repair; exhaust; exhaustion; experimental study; gamma secretase; genetic manipulation; horizontal cell; in vivo; in vivo evaluation; inhibitor; neurogenesis; neuron regeneration; notch protein; novel; novel strategies; olfactory neurogenesis; olfactory sensory neurons; post-transplant; prospective; receptor; respiratory; small molecule; small molecule inhibitor; stem cell function; stem cells; tissue culture; transcription factor

PROJECT SUMMARY The capacity of the adult olfactory epithelium (OE) for renewing and regenerating the population of sensory neurons depends on the persistence and proper function of stem cells. Presbyosmia is accompanied by pathological changes due to the disordering and eventual depletion of the normally active olfactory stem and progenitor cells, namely globose basal cells (GBCs). In this setting, the reserve stem cells, namely the horizontal basal cells (HBCs), remain dormant despite the neurogenic exhaustion and disappearance of GBCs. In contrast, if the OE is damaged by an olfactotoxin, the HBCs activate and contribute to the repair of the epithelium. The results of HBC activation are strongly context-dependent and do not always capacitate the rejuvenation of neurogenesis. That failure can occur in rodents as a consequence of either experimental lesion or aging/neurogenic exhaustion. Substantial indirect evidence suggests the same is true of the human OE. A therapeutic strategy that directs HBCs down the pathway toward neuronal regeneration in the setting of an exhausted OE offers possibly the best approach for treating age-related olfactory dysfunction. We have demonstrated that the TF p63 is the master switch that regulates HBC activation – a precipitous decline in p63 levels is necessary and sufficient for activation. We are testing factors that are known to enhance neuronal differentiation (Aim 1) or suppress non-neuronal differentiation (Aim 2). We will use quantitative proteomics to define the secretome of olfactory stromal cells, which will nominate candidate mechanisms for the intricate intercommunication between epithelium and cells of the lamina propria that regulates neurogenesis. We will test their efficacy in vitro using our newly developed HBC culture system by quantifying the proportion of neurons that emerge after activation by the various factors. Then we will validate the in vitro effect by transplanting the activated/factor-treated HBCs into the lesioned OE, which provides a neutral environment in which all neuronal and non-neuronal fates are available to either endogenous HBCs or ones that have been cultured. We will test efficacy on both mouse and human HBCs, and we will use a novel xenotransplantation assay to assess the consequences for human HBCs in vivo. When completed, we will have achieved a much more thorough understanding of the process by which HBCs are directed toward neuronal differentiation so that they might rejuvenate neurogenesis. That understanding of mechanism both in mouse, where genetic manipulations offer profound analytic power, and in humans will advance our efforts aimed at identifying therapeutic strategies for alleviating olfactory sensory dysfunction, particularly the sensory loss which accompanies aging.

项目摘要 成人嗅觉上皮（OE）的能力，以更新和再生感官的种群 神经元取决于干细胞的持久性和正常功能。长期症是由 正常活跃的嗅觉茎和最终部署导致的病理变化 祖细胞，即球体碱性细胞（GBC）。在这种情况下，储备干细胞，即 尽管具有神经源性疲劳和GBC消失，但水平基线细胞（HBC）仍处于休眠状态。 相比之下，如果OE被橄榄毒素损坏，HBC会激活并有助于修复 上皮。 HBC激活的结果非常依赖上下文，并且并不总是能使 神经发生的复兴。由于任何一种实验，这种失败可能在啮齿动物中发生 病变或衰老/神经源性疲惫。大量间接证据表明，人类也是如此 OE。一种治疗策略，该策略将HBC引导在神经元再生的途径中 疲惫的OE为治疗与年龄相关的嗅觉功能障碍提供了最佳方法。我们有 证明TF p63是调节HBC激活的主开关 - p63的精确下降 水平是必要的，足以激活。我们正在测试已知可以增强神经元的因素 分化（AIM 1）或抑制非神经元分化（AIM 2）。我们将使用定量蛋白质组学进行 定义嗅觉基质细胞的分泌组，该细胞将提名复杂的候选机制 调节神经发生的上皮和细胞的细胞之间的互通。我们将 使用我们新开发的HBC培养系统在体外测试其效率，通过量化比例 各种因素激活后出现的神经元。然后，我们将通过 将激活/因子处理的HBC移植到病变的OE中，该OE提供了中性环境 所有神经元和非神经元的命运都可以用于内源性HBC或已有的命运 培养。我们将测试小鼠和人类HBC的效率，我们将使用新型的异种移植术 测定评估体内人类HBC的后果的测定。完成后，我们将取得很多成就 更彻底地了解HBC针对神经元分化的过程，因此 它们可能会恢复神经发生。对遗传学的机制的理解 操纵提供了深刻的分析能力，在人类中，我们将促进我们旨在识别的努力 减轻嗅觉感觉功能障碍的治疗策略，尤其是感官损失 伴随着衰老。