Investigating the effects of psychosocial stress on laryngeal microbiology and epithelial barrier integrity

摘要 心理社会应激导致喉部改变的生物学机制仍未得到充分研究。 在有嗓音问题的患者中，有25%同时出现心理应激症状。从一个 从生物学的角度来看，心理社会压力会对这两个微生物群落产生有害后果 和上皮屏障的完整性，如报告的，在其他粘膜器官，如肠道和胃。首先， 与肠道相比，喉部微生物群具有不同的微生物组成。这一点有多么明显 作曲是否受到心理社会压力的影响仍不清楚。独立地，压力诱导的变化 上皮通透性(如上皮细胞增殖减少和粘液厚度减少)可能 使喉部容易受到进一步感染。这项提议笼统地质疑心理社会压力是否 可导致微生物失调和受损的上皮屏障完整性。在我们提议的实验中，小鼠 将被分配到无压力或有压力的小组。心理社会压力将通过一种有效的慢性 拘束应激协议。特定目标1将确定喉部微生物在慢性、 心理社会压力暴露。我们假设心理社会压力会导致微生物减少 特定细菌分类群的多样性和丰度、变化的组成和不同的丰度，以及不同的 预测了喉部微生物区系的功能分布。为了获得喉部微生物区系的结果，16S rRNA 将对从小鼠喉部组织中提取的细菌进行完整的测序。特定目标2将 描述慢性心理社会应激后声带上皮完整性的变化。我们假设 心理社会压力会导致细菌移位增加，粘液厚度减少， 上皮细胞增殖，上皮细胞连接完整性降低，粘蛋白基因表达改变 声带。免疫组织化学/免疫荧光和定量聚合酶链式反应将用于描述 喉上皮的心理社会压力。综上所述，我们假设心理社会压力会 同时导致微生物失调，改变喉部上皮屏障的完整性。这两个项目的数据 AIMS将为鉴定和操纵特定的细菌门以描绘宿主微生物奠定基础 喉部心理社会应激的上皮相互作用。

ABSTRACT Biological mechanisms underlying psychosocial stress-induced changes to the larynx remain understudied. Psychological stress symptomology is concurrently reported in 25% of patients with voice problems. From a biological perspective, psychosocial stress can have deleterious consequences on both microbial communities and epithelial barrier integrity, as reported, in other mucosal organs such as the gut and stomach. First, the laryngeal microbiome has a distinct microbial composition when compared to the gut. How this distinct composition is influenced by psychosocial stress remains unknown. Independently, stress-induced changes to epithelial permeability (such as reduced epithelial proliferation and reduced mucus thickness) may potentially leave the larynx susceptible to further infection. This proposal broadly questions whether psychosocial stress can lead to microbial dysbiosis and compromised epithelial barrier integrity. In our proposed experiments, mice will be assigned to unstressed or stressed groups. Psychosocial stress will be induced via a validated chronic restraint stress protocol. Specific Aim 1 will determine laryngeal microbial adaptations following chronic, psychosocial stress exposure. We hypothesize that psychosocial stress will result in reduced microbial diversity and abundance, altered composition and differential abundance of specific bacterial taxa, and distinct predicted functional profiles in the laryngeal microbiota. To obtain laryngeal microbiota outcomes, 16s rRNA sequencing will be used completed on bacteria extracted from murine laryngeal tissue. Specific Aim 2 will delineate changes in vocal fold epithelial integrity post-chronic psychosocial stress. We hypothesize that psychosocial stress will result in increased bacterial translocation, decreased mucus thickness, reduced epithelial proliferation, reduced epithelial cell junction integrity, and altered gene expression of mucins in the vocal folds. Immunohistochemistry/ Immunofluorescence and qPCR will be used to delineate consequences of psychosocial stress on the laryngeal epithelium. Taken together, we hypothesize that psychosocial stress will concomitantly lead to microbial dysbiosis and alter epithelial barrier integrity in the larynx. Data from these two aims will lay the groundwork for identifying and manipulating specific bacterial phyla to delineate host-microbial epithelial interactions of psychosocial stress in the larynx.