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.

项目概要 成人嗅觉上皮 (OE) 更新和再生感觉群体的能力 老花症的神经元依赖于干细胞的持续存在和正常功能。 由于正常活跃的嗅觉干的紊乱和最终耗竭而引起的病理变化 祖细胞，即球形基底细胞（GBC）在这种情况下，储备干细胞，即 尽管神经源性衰竭和 GBC 消失，但水平基底细胞 (HBC) 仍处于休眠状态。 相反，如果 OE 被嗅毒素损坏，HBC 就会激活并有助于修复 HBC 激活的结果强烈依赖于环境，并且并不总是使 HBC 具有能力。 啮齿类动物的神经发生的恢复可能是由于任一实验的结果而发生的。 大量间接证据表明，人类的情况也是如此。 OE。一种在以下情况下引导 HBC 沿着神经元再生途径的治疗策略： 疲惫的嗅觉可能是治疗与年龄相关的嗅觉功能障碍的最佳方法。 TF p63 是调节 HBC 激活的主开关 - p63 急剧下降 水平对于激活是必要且充分的，我们正在测试已知可增强神经的因素。 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进行。 定义嗅觉基质细胞的分泌组，这将为复杂的机制提名候选机制 上皮细胞和固有层细胞之间调节神经发生的相互通讯。 使用我们新开发的 HBC 培养系统，通过量化 然后我们将通过各种因素激活后出现的神经元来验证体外效果。 将经过活化/因子处理的 HBC 移植到受损的 OE 中，从而在其中提供中性环境 所有神经元和非神经元的命运都可用于内源性 HBC 或已被 我们将测试对小鼠和人类 HBC 的功效，并且我们将使用一种新型异种移植。 评估人类 HBC 体内影响的测定完成后，我们将取得很大成果。 更透彻地了解 HBC 定向神经分化的过程，以便 他们可能会恢复小鼠神经发生的机制，其中遗传。 操纵提供了深刻的分析能力，并且在人类中将推进我们旨在识别的努力 缓解嗅觉感觉功能障碍的治疗策略，特别是感觉丧失 伴随着衰老。