Effect of vocal fold injury on laryngeal muscle dysfunction

摘要 嗓音障碍影响了数百万美国人，造成了巨大的经济损失。关于以下方面的决定 治疗通常主要基于两种思维：粘膜损伤和肌肉功能障碍。 然而，喉肌肉功能障碍常常伴随着喉粘膜病理，并且可能两者兼而有之 使役的和反应性的。这一建议是体内和体外方法的创新组合，以 研究暂时性和持续性声带粘膜损伤对潜在功能的影响， 声带的主要肌肉--甲颧肌的结构和机械反应。这个 这项研究的长期目标是提供关于两者之间复杂相互作用的机械性见解 声带粘膜层和肌层，从而有助于临床模式和分类的转变 嗓音障碍，但更重要的是，导致开发新的、有针对性的治疗策略 数以百万计的与声音相关的残疾患者。这项工作的中心假设是声带受伤。 折叠在声带粘膜和底层肌肉之间引起复杂的相互作用，因此 导致喉部肌肉结构和功能的改变。 这项拟议的研究有两个具体目标。目标1是确定功能、结构和 在已建立的活体啮齿动物模型中，喉肌对声带损伤的机械性反应。两个深度 声带损伤、仅粘膜浅损伤或粘膜和下层肌肉深度损伤， 将在雄性和雌性大鼠中创建，以确定声带损伤对a)肌肉功能的时间进程 通过分析超声发声的声学复杂性，b)肌肉的机械反应通过 蛋白质组分析，以及c)在大数据蛋白质组询问中确定的特定蛋白质水平的变化。 损伤深度将通过肌肉组织学来确认。目标2是描述和量化信号之间的关系 基础状态和刺激状态下体外培养的声带成纤维细胞、成肌细胞和炎性细胞 (例如，受伤)情况。体外实验将检验肝纤维化和炎症的影响 声带固有层成纤维细胞和喉内肌成肌细胞的介体 通过共培养和条件培养液实验，询问两种细胞类型之间的独特相互作用。 这些数据将提供关于发音困难和发音周期的关键的机械性和转化性的见解 为药物干预提供潜在靶点。 提出的研究具有重要意义，因为这种基础性的机制洞察力将优化 为数百万与嗓音相关的残疾患者提供治疗。这项提议在概念上是创新的， 提出新颖、尖端的方法，并整合多个实验室和 特色菜。

ABSTRACT Voice disorders affect millions of Americans at substantive economic cost. Decisions regarding treatment are often primarily based on dichotomous thinking: mucosal injury versus muscle dysfunction. However, laryngeal muscle dysfunction often accompanies laryngeal mucosal pathologies and may be both causative and reactive. This proposal is an innovative combination of in vivo and in vitro methodologies to investigate the effect of both transient and persistent vocal fold mucosal injury on the underlying function, structure, and mechanistic response of the thyroarytenoid muscle, the primary muscle of the vocal fold. The long-term goal of this research is to provide mechanistic insight regarding the complex interactions between the mucosal and muscular vocal fold layers, thereby contributing to a shift in clinical schema and classification of voice disorders, but more importantly, lead to the development of novel, targeted therapeutic strategies for the millions of patients with voice-related disability. The central hypothesis of this work is injury to the vocal folds elicits complex interactions between the vocal fold mucosa and underlying muscle, which consequently results in altered laryngeal muscle structure and function. The proposed research has two specific aims. Aim 1 is to determine functional, structural, and mechanistic laryngeal muscle responses to vocal fold injury in an established in vivo rodent model. Two depths of vocal fold injuries, shallow injury of the mucosa only or deep injury of the mucosa and underlying muscle, will be created in male and female rats to determine the time course of vocal fold injury on a) muscle function via analysis of ultrasonic vocalization acoustic complexity, b) mechanistic response of the muscle via proteomic analysis, and c) changes in specific protein levels identified in the big-data proteomic interrogation. Injury depths will be confirmed via muscle histology. Aim 2 is to characterize and quantify signaling between fibroblasts and myoblasts as well as inflammatory cells from the vocal folds in vitro under basal and stimulated (e.g., injured) conditions. In vitro experimentation will examine the effects of fibrotic and inflammatory mediators on fibroblasts from the vocal fold lamina propria and myoblasts from intrinsic laryngeal muscle to interrogate unique interactions between the two cell types via co-culture and conditioned media experiments. These data will provide critical mechanistic and translational insight regarding the cycle of dysphonia and provide potential targets for pharmacological interventions. The proposed research is significant because this foundational mechanistic insight will optimize treatments for millions of patients with voice-related disability. The proposal is conceptually innovative, proposes novel, cutting-edge methodologies, and coalesces expertise across multiple laboratories and specialties.