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

摘要 心理社会压力引起的喉部变化的生物学机制仍然研究不足。 心理压力神经病学同时报告在25%的患者的声音问题。从 从生物学的角度来看，心理社会压力可能对两种微生物群落都产生有害影响。 和上皮屏障完整性，如所报道的，在其它粘膜器官如肠和胃中。一是 与肠道相比，喉部微生物组具有不同的微生物组成。这种独特的 社会心理压力对儿童组成的影响尚不清楚。独立地，压力引起的变化， 上皮渗透性（例如上皮增殖减少和粘液厚度减少）可能潜在地 使喉部容易受到进一步感染这一提议广泛质疑社会心理压力是否 可导致微生物生态失调和损害上皮屏障完整性。在我们提出的实验中， 将被分配到无压力组或有压力组。心理社会压力将通过一种有效的慢性 束缚应激方案具体目标1将确定慢性， 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， 多样性和丰度，改变了特定细菌分类群的组成和差异丰度， 预测喉部微生物群的功能概况。为了获得喉部微生物群结果，16 s rRNA 测序将用于从鼠喉组织中提取的细菌。具体目标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，并改变了粘蛋白的基因表达， 声带免疫组织化学/免疫荧光和qPCR将用于描述 对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 同时导致微生物生态失调并改变喉中的上皮屏障完整性。这两个数据 目标将为识别和操纵特定细菌门以描绘宿主微生物奠定基础 心理社会压力在喉上皮的相互作用。

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.