Evaluating the role of epithelial basal cells in laryngeal homeostasis and disease development

摘要 喉部需要保护，以抵御在履行其职责期间每天遇到的约2500万次吸入性侮辱 正常的功能，包括呼吸、咳嗽、吞咽，以及人类的发声。一种上皮细胞 在喉部表面形成关键的第一道保护性屏障。在正常情况下，上皮细胞 依赖于体内平衡的再生机制。上皮的基底细胞层被认为是 代表形成和再生分化上皮的祖细胞的储存库。 然而，喉部的组成和组织会发生病理变化或重塑。 上皮细胞对吸入侮辱，特别是香烟烟雾(CS)的反应。重塑会影响心脏的功能 上皮起到有效屏障的作用，使组织易于发生疾病。尽管 CS诱导的喉部疾病中上皮重塑的发生率和可能的重要性，其病因尚不清楚 明白了。这是由于对喉上皮细胞的调控机制缺乏基本的了解。 与CS相关的动态平衡再生和永久性组织重塑。具体地说，人们对 不列颠哥伦比亚省种群在正常的上皮内稳态、损伤或疾病条件下的生物学。稀少的 喉癌生物学的研究是预防和治疗喉癌的主要障碍。 治疗CS病的声带。为了解决这一未得到满足的需求，我们建议测试BC 在正常喉上皮的形成和维持以及CS诱导的BC改变中起关键作用 生物学与上皮重塑和人类疾病的发展有关。我们将检验这一假设 用正常小鼠模型(AIM-1)和体内、体外CS暴露模型(AIM-2)研究喉部结构 分化上皮的BC特性、原代培养和生理相关的三维 上皮细胞培养系统。我们还建议评估我们在小鼠体内和体外的研究结果之间的关系。 通过评估患者喉部标本中BC的生物学特性来评估人类状况 Reinke‘s水肿，一种常见的CS引起的喉部疾病(AIM 3)。这些目标的实现将证明 BC在喉上皮生物学中具有中心和不可替代的作用-它们作为 正常的上皮和BC生物学的改变与疾病的发展有关。调查结果将会有 重大的理论和临床影响，并将为潜在的机制提供新的见解 喉和声带上皮再生。靶向介导CS诱导的特定生物通路 BC生物学中的错乱可能代表着创新的治疗目标，以防止发育和 CS和其他吸烟引起的嗓音障碍的进展。

ABSTRACT The larynx requires protection from the ~25 million inhaled insults encountered daily during fulfillment of its normal functions including breathing, coughing, swallowing, and in humans, voice production. An epithelium forms a critical first protective barrier on the surface of the larynx. Under normal conditions, the epithelium relies on homeostatic regenerative mechanisms. The basal cell (BC) layer of the epithelium is believed to represent a reservoir of progenitor cells for the formation and regeneration of differentiated epithelium. However, pathological changes, or remodeling, occurs in the composition and organization of the laryngeal epithelium in response to inhaled insults, notably cigarette smoke (CS). Remodeling affects the ability of the epithelium to function as an effective barrier and predisposes the tissue to disease development. Despite the prevalence and likely importance of epithelial remodeling in CS-induced laryngeal disease, its etiology is poorly understood. This is due to a lack of basic knowledge of the mechanisms which dictate laryngeal epithelial homeostatic regeneration and permanent tissue remodeling linked to CS. Specifically, little is known about the biology of the BC population in normal epithelial homeostasis, injured, or diseased conditions. A paucity of research on laryngeal BC biology serves as a major barrier in the development of therapies to prevent and treat CS-diseased vocal folds. To address this unmet need, we propose to test the central hypothesis that BC are critical to the formation and maintenance of normal laryngeal epithelium and CS-induced alterations in BC biology are associated with epithelial remodeling and human disease development. We will test this hypothesis using a normal mouse model (AIM 1) and in vivo and in vitro models of CS exposure (AIM 2) to study laryngeal BC properties in differentiated epithelium, primary culture, and physiologically relevant three-dimensional epithelial cell culture systems. We also propose to evaluate how our in vivo and in vitro findings in mice relate to the human condition by evaluating the biology of BC in human laryngeal specimens from patients with Reinke’s edema, a common CS-induced laryngeal disease (AIM 3). Completion of these aims will demonstrate that BC have a central and irreplaceable role in laryngeal epithelial biology- they function as progenitor cells for normal epithelium and alterations in BC biology are implicated in disease development. Findings will have significant theoretical and clinical impact and will provide novel new insights into the mechanism underlying laryngeal and vocal fold epithelial regeneration. Targeting specific biologic pathways that mediate CS-induced derangements in BC biology may represent innovative therapeutic targets to prevent development and progression of CS- and other smoke-mediated voice disorders.