Vocal fold epithelial barrier and proliferation in laryngeal papillomatosis

摘要 复发性呼吸道乳头状瘤病（RRP）是一种高度病态的上呼吸道疾病， 引起人乳头瘤病毒（HPV）6型和11型。RRP的特点是快速增长的集群 喉部和上呼吸道的上皮病变虽然病变在组织学上是良性的， 疾病，并可能导致危及生命的气道阻塞，特别是在儿童中。反复外科 每隔几个月切除一次病变以维持呼吸可能会导致声带疤痕， 进一步损害语音功能。疫苗只是预防性的。没有治愈已建立的HPV感染或 对于RRP。由于缺乏临床前模型，RRP研究受到限制。乳头瘤病毒需要分层 完成其生命周期。临床上，RRP进展包括复发和消退。 病变疾病的这些方面最好使用动物模型来捕捉， 乳头瘤病毒也是物种特异性的。最近发现的小鼠乳头状瘤病毒MmuPV 1， 创造了新的机会，在遗传修饰的实验室动物中模拟乳头瘤病毒感染。 然而，迄今为止，还没有关于MmuPV 1在声带和/或喉中的研究。这项工作的总体目标是 利用一种新的鼠乳头状瘤病毒， 喉乳头状瘤病模型。目的1将探讨机械损伤在MmuPV 1疾病中的作用 使用疾病负荷和病毒载量的纵向测量，以及 在创伤期间的多个时间点定位病毒RNA和组织内的感染性颗粒的组装 治愈同时，目标2将表征上皮形态、屏障完整性和细胞增殖的变化。 在整个伤口愈合过程中，由乳头瘤病毒感染诱导增殖和死亡。我们的核心假设 上皮损伤将促进病毒感染，而感染将破坏正常上皮完整性， 细胞周转率通过阐明声带损伤在喉乳头状瘤病发病中的作用和初步疗效， 声带上皮乳头状瘤病，完成我们的具体目标将产生重大影响， 咽喉学和病毒学。这一提议将产生一种新的RRP临床前模型， 研究疾病的持续性和复发，包括免疫系统的作用，并最终将 用于开发针对这种棘手和毁灭性疾病的创新预防和治疗策略。

ABSTRACT Recurrent respiratory papillomatosis (RRP) is a highly morbid upper airway disease caused by non-cancer- causing human papillomavirus (HPV) types 6 and 11. RRP is characterized by quickly growing clustered epithelial lesions in the larynx and upper airway. Although lesions are histologically benign, they cause voice disorders and can result in life-threatening airway obstruction, particularly in children. Repeated surgical excision of lesions required as often as every few months to maintain breathing can cause vocal fold scar and further impair voice function. Vaccination is preventive only. There is no cure for established HPV infection or for RRP. RRP research has been limited by the lack of a preclinical model. Papillomaviruses require layered epithelium to complete their life cycle. Clinically, RRP progression includes regression and recurrence of lesions. These aspects of disease are best captured using animal models, which have been limited since papillomaviruses are also species-specific. The recent discovery of murine papillomavirus MmuPV1 has created novel opportunities to model papillomavirus infection in genetically modifiable laboratory animals. However, there are no studies of MmuPV1 in vocal folds and/or larynx to date. The overall goal of this work is to define the interaction between vocal fold epithelial injury and papillomavirus infection using a novel murine model of laryngeal papillomatosis. Aim 1 will investigate the role of mechanical injury on MmuPV1 disease progression in murine larynx using longitudinal measures of disease burden and viral load, as well as localizing viral RNA and assembly of infectious particles within tissue at multiple timepoints during wound healing. In parallel, Aim 2 will characterize changes in epithelial morphology, barrier integrity, and cell proliferation and death induced by papillomavirus infection throughout wound healing. Our central hypothesis is that epithelial injury will facilitate viral infection, and that infection will disrupt normal epithelial integrity and cell turnover. By clarifying the role of vocal fold injury in onset of laryngeal papillomatosis and the initial effects of papillomatosis on vocal fold epithelium, completion of our specific aims will have a significant impact on laryngology and virology. This proposal will result in a novel preclinical model of RRP that will facilitate further study of disease persistence and recurrence, including the role of the immune system, and will ultimately be used to develop innovative prevention and treatment strategies for this intractable and devastating disease.