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: 10642890
• 负责人: JAMES E. SCHWOB
• 金额: $ 20.63万
• 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 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10642890
• 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 Cells; Epithelium; 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; 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; cofactor; directed differentiation; efficacy testing; efficacy validation; epithelial injury; epithelial repair; exhaust; exhaustion; experimental study; genetic manipulation; horizontal cell; in vivo; in vivo evaluation; inhibitor; mortality; neurogenesis; neuron regeneration; notch protein; novel; novel strategies; olfactory neurogenesis; olfactory sensory neurons; post-transplant; prospective; receptor; secretase; small molecule inhibitor; stem; stem cell function; stem cells; synergism; 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.

项目总结 成体嗅觉上皮(OE)更新和再生感觉群的能力 神经元依赖于干细胞的持久性和正常功能。老年性嗅觉障碍伴有 由于正常活动的嗅干紊乱和最终枯竭而引起的病理变化 祖细胞，即球状基底细胞(GBCs)。在这种情况下，储备干细胞，即 水平基底细胞(HBCs)尽管神经源性衰竭和消失，但仍处于休眠状态。 相反，如果嗅觉毒素损伤了OE，HbCs就会激活，并有助于修复 上皮组织。HBC激活的结果强烈地依赖于上下文，并且并不总是使 神经再生的恢复活力。这种失败可能发生在啮齿类动物身上，作为实验的结果之一 损伤或衰老/神经性疲惫。大量间接证据表明，人类也是如此。 大江南北。一种治疗策略，引导HbCs沿着通路走向神经元再生 精疲力竭的OE可能是治疗年龄相关性嗅觉功能障碍的最佳方法。我们有 表明转铁蛋白p63是调节HbC激活的主开关--p63的急剧下降 水平是激活所必需的，也是充分的。我们正在测试已知的增强神经元的因子。 分化(目标1)或抑制非神经元分化(目标2)。我们将使用定量蛋白质组学来 定义嗅觉基质细胞的分泌组，它将提名复杂的 调节神经发生的固有层上皮和细胞之间的相互联系。我们会 使用我们最新开发的HBC培养系统，通过量化 被各种因素激活后出现的神经元。然后我们将通过以下方式验证体外效应 将活化的/经因子处理的HBCs移植到受损的OE中，这在 所有神经元和非神经元的命运对内源性或已经被 有教养的。我们将在小鼠和人的Hbc上测试疗效，我们将使用一种新的异种移植 在活体内评估对人HBCs的影响。建成后，我们将取得更大的成就 更深入地了解HbC定向向神经元分化的过程 它们可能会使神经再生恢复活力。对小鼠机制的理解，在那里基因 操纵提供了深刻的分析能力，在人类身上，将推动我们旨在识别 减轻嗅觉功能障碍，特别是嗅觉丧失的治疗策略 伴随着年龄的增长。