Transcriptomic analysis of vulvovaginal candidiasis identifies a role for the NLRP3 inflammasome.
Transcriptomic analysis of vulvovaginal candidiasis identifies a role for the NLRP3 inflammasome.
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DOI：
10.1128/mbio.00182-15
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发表时间：
2015-04-21
期刊：
影响因子：
6.4
通讯作者：
Peters BM
中科院分区：
文献类型：
--
作者：
Bruno VM;Shetty AC;Yano J;Fidel PL Jr;Noverr MC;Peters BM
外阴阴道念珠菌病（VVC），最常见的白色念珠菌引起的治疗，代表了一个显着的未满足的临床需求。白色念珠菌，作为一种阴道和致病生物体，与阴道环境有着复杂且知之甚少的相互作用。了解这种关系的复杂性质对于开发迫切需要的治疗有症状感染的疗法是必要的。使用转录组测序（RNA-seq），我们的特点是早期小鼠阴道和真菌的转录组的有机体在VVC。对感染和未感染小鼠之间差异表达的宿主基因的网络分析预测了先前与VVC无关的几种信号传导途径的激活或抑制，包括NLRP 3炎性体激活。用白色念珠菌对Nlrp3 −/−小鼠进行阴道内激发，结果显示阴道灌洗液中多形核白细胞（PMN）、警报素和炎性细胞因子（包括白细胞介素-1 β（IL-1 β）（VVC免疫发病机制的标志）水平严重降低。野生型（WT）小鼠阴道内给予格列本脲（一种NLRP 3炎性体的强效抑制剂），可将PMN浸润和IL-1 β降低至与Nlrp 3 −/−小鼠中观察到的水平相当。此外，白色念珠菌基因的RNA-seq分析表明菌丝相关的分泌型乙酰基蛋白酶4、5和6（SAP 4 - 6）的稳健表达，其是已知的炎性小体激活剂。尽管定植与WT株相似，但Δ SAP 4 - 6三联突变体和Δ SAP 5单突变体在阴道内激发期间诱导的PMN流入和IL-1 β显著减少。我们的研究结果表明，炎性小体在VVC的免疫发病机制中发挥了新的作用，并暗示菌丝相关的SAP是外阴阴道念珠菌病期间白色念珠菌的主要毒力决定因素。阴道炎，最常见的由真菌白色念珠菌引起，导致所有育龄妇女的生活质量问题。最近的研究表明，阴道炎是由宿主先天免疫控制的免疫病理反应引起的，尽管解释机制尚未确定。使用全面的基因组学，免疫学和药理学方法，我们已经阐明了NLRP 3炎性体作为一个重要的分子机制，有助于宿主免疫病理学。我们还证明了白色念珠菌菌丝相关的分泌型β-淀粉酰蛋白酶（更具体地说，SAP 4 - 6和SAP 5）有助于疾病的免疫病理学。最终，这项研究增强了我们对阴道粘膜宿主和真菌之间复杂相互作用的了解，并为炎性小体治疗症状性外阴阴道念珠菌病提供了原理性证据。
Treatment of vulvovaginal candidiasis (VVC), caused most frequently by Candida albicans, represents a significant unmet clinical need. C. albicans, as both a commensal and a pathogenic organism, has a complex and poorly understood interaction with the vaginal environment. Understanding the complex nature of this relationship is necessary for the development of desperately needed therapies to treat symptomatic infection. Using transcriptome sequencing (RNA-seq), we characterized the early murine vaginal and fungal transcriptomes of the organism during VVC. Network analysis of host genes that were differentially expressed between infected and naive mice predicted the activation or repression of several signaling pathways that have not been previously associated with VVC, including NLRP3 inflammasome activation. Intravaginal challenge of Nlrp3−/− mice with C. albicans demonstrated severely reduced levels of polymorphonuclear leukocytes (PMNs), alarmins, and inflammatory cytokines, including interleukin-1β (IL-1β) (the hallmarks of VVC immunopathogenesis) in vaginal lavage fluid. Intravaginal administration of wild-type (WT) mice with glyburide, a potent inhibitor of the NLRP3 inflammasome, reduced PMN infiltration and IL-1β to levels comparable to those observed in Nlrp3−/− mice. Furthermore, RNA-seq analysis of C. albicans genes indicated robust expression of hypha-associated secreted aspartyl proteinases 4, 5, and 6 (SAP4–6), which are known inflammasome activators. Despite colonization similar to that of the WT strain, ΔSAP4–6 triple and ΔSAP5 single mutants induced significantly less PMN influx and IL-1β during intravaginal challenge. Our findings demonstrate a novel role for the inflammasome in the immunopathogenesis of VVC and implicate the hypha-associated SAPs as major C. albicans virulence determinants during vulvovaginal candidiasis. Vaginitis, most commonly caused by the fungus Candida albicans, results in significant quality-of-life issues for all women of reproductive age. Recent efforts have suggested that vaginitis results from an immunopathological response governed by host innate immunity, although an explanatory mechanism has remained undefined. Using comprehensive genomic, immunological, and pharmacological approaches, we have elucidated the NLRP3 inflammasome as a crucial molecular mechanism contributing to host immunopathology. We have also demonstrated that C. albicans hypha-associated secreted aspartyl proteinases (SAP4–6 and SAP5, more specifically) contribute to disease immunopathology. Ultimately, this study enhances our understanding of the complex interplay between host and fungus at the vaginal mucosa and provides proof-of-principle evidence for therapeutic targeting of inflammasomes for symptomatic vulvovaginal candidiasis.