A novel role for the inflammasome in the immunopathogenesis of Candida vaginitis

炎症小体在念珠菌阴道炎免疫发病机制中的新作用

• 批准号: 9206120
• 负责人: Brian M Peters
• 金额: $ 10.8万
• 依托单位: UNIVERSITY OF TENNESSEE HEALTH SCI CTR
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-01-15 至 2018-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9206120
• 关键词: Affect; Age; Alternative Therapies; Animals; Antifungal Agents; Aspartic Endopeptidases; Bacterial Adhesins; Belief; Biological; Burning Pain; CASP1 gene; Candida; Candida albicans; Career Mobility; Chronic; Clinical; Consult; Cytokine Signaling; Data; Defect; Development; Disease; Epithelial; Epithelium; Faculty; Female of child bearing age; Foundations; Funding; Funding Opportunities; Genetic; Genetic Transcription; Glyburide; Goals; Health; Histologic; Human Volunteers; Immune; Immune response; Immunity; Immunologics; In Vitro; Infection; Inflammasome; Inflammation; Inflammatory; Inflammatory Response; Innate Immune Response; Interleukin-1 beta; Interleukins; K22 Award; Knockout Mice; Knowledge; Lactate Dehydrogenase; Lead; Liquid substance; Manuscripts; Measurement; Mediating; Mediator of activation protein; Medical; Mentors; Mission; Molecular; Morphogenesis; Mucous Membrane; Mus; National Institute of Allergy and Infectious Disease; Neutrophil Infiltration; Oral; Oral mucous membrane structure; Outcome; Output; Pain; Peptide Hydrolases; Pharmacology; Positioning Attribute; Predisposition; Preventive therapy; Process; Production; Pruritus; Publications; Publishing; Quality of life; Receptor Signaling; Recruitment Activity; Recurrence; Redness; Research; Research Personnel; Role; Severities; Signal Pathway; Signal Transduction; Signaling Molecule; Surface; Symptoms; Testing; Tissues; Translational Research; Vagina; Vaginal Douching; Vaginal Itching; Vaginitis; Virulence; Virulence Factors; Vulvovaginal Candidiasis; Woman; Work; Yeasts; adaptive immunity; career; combat; cost; cytokine; design; experience; faculty research; fungus; immunopathology; in vivo; inflammatory marker; innovation; member; migration; mouse model; neutrophil; new therapeutic target; novel; novel strategies; novel therapeutic intervention; overexpression; pathogen; professor; programs; public health relevance; reproductive; response; skills; tenure track; treatment strategy

DESCRIPTION (provided by applicant): Candida albicans, the primary causative agent of fungal vaginitis, will affect 75% of all women of reproductive age at least once in their lifetime. Static activity of commonly deployed antifungals and chronic infections are of growing concern; therefore, alternative therapies are crucially needed. Despite long-held beliefs that susceptibility to Candida vaginitis resulted from defects in adaptive immunity, it has recently been shown that the clinical immunopathology (i.e. itching, burning, pain, discomfort, redness of the vaginal mucosa) associated with Candida vaginitis is triggered by robust recruitment of polymorphonuclear leukocytes (PMN) to the vaginal epithelium. Using a mouse model of vaginitis, the S100 alarmins were identified as key signaling molecules in in this hallmark PMN response. However, recent studies using mice deficient in S100 alarmin production still responded with vigorous PMN recruitment when challenged with C. albicans. Therefore, a fundamental gap in knowledge still exists as to the alternative and/or complementary mechanism(s) by which neutrophils are recruited to the vaginal epithelium during Candida vaginitis. One possible process by which inflammation can be triggered at epithelial surfaces is by activation of the inflammasomes, a group of cytosolic cellular receptors that signal and initiate innate immune responses. Therefore, the objective of this proposal is to 1) demonstrate that the Nlrp3 inflammasome is a key early mediator of the innate inflammatory response and to 2) identify specific hypha-associated C. albicans virulence factors that result in inflammasome activation in the vaginal epithelium. Guided by strong preliminary data, these aims will test our central hypothesis that activation of the inflammasome induced by the C. albicans yeast-to-hypha transition during vaginitis potentiates exacerbated neutrophil influx and pro-inflammatory cytokine signaling in the vaginal epithelium. All aims will utilize a well-established mouse model of vaginitis to test our hypotheses. Under the first aim, genetic or pharmacologic blockade of discrete targets in the inflammasome signaling cascade will be utilized to elucidate signaling pathways contributing to Candida vaginitis. Successful disruption of inflammasome-dependent signaling will be assessed via measurement of inflammatory markers (PMNs, Interleukin-1b, S100 alarmins). In the second aim, morphogenesis-competent C. albicans strains deficient in specific hypha-associated virulence factors (secreted proteinases or adhesins) or strains locked in the yeast form but over-expressing these key identified hyphal virulence factors will be utilized to assess their required role for generating robust innate immune responses in vivo. This approach is innovative because it examines both host and fungal processes required for immune responses associated with Candida vaginitis and seeks to identify inflammasome function in the vaginal mucosa. This proposed work is significant because it will ultimately aid in the identification of novel signaling pathways that may serve as new therapeutic targets for such infections. This work will also provide a strong foundation on which the candidate can attain an independent academic research program. The candidate's immediate career goal is to successfully transition from a non-tenure track junior faculty member to a tenure-track Assistant Professor position studying the host innate immune response and molecular mechanisms associated with fungal vaginitis. His long-term career goal is to attain a tenured research faculty position to direct and maintain a well-funded, translational research team that studies innate immune mechanisms to fungal pathogens at the mucosal interface. In order to attain these career goals, aside from conducting the research described above, the candidate will assemble and maintain a supportive mentoring/consulting team that will aid in the successful completion of the research, hone the development of grantsmanship skills leading to effective funding opportunities, and accelerate the publication of high-impact data. Thus, these career goals fulfill the NIAID's mission to identify novel therapeutic approaches to combat fungal disease and to support junior investigators in making a career transition to independent faculty status under the K22 award mechanism.

描述（由申请人提供）： 白色念珠菌是真菌性阴道炎的主要病原体，75% 的育龄妇女一生中至少会感染一次。常用抗真菌药物和慢性感染的静态活性日益受到关注；因此，迫切需要替代疗法。尽管长期以来人们认为对念珠菌性阴道炎的易感性是由以下缺陷引起的： 适应性免疫，最近表明，与念珠菌阴道炎相关的临床免疫病理学（即阴道粘膜瘙痒、灼烧感、疼痛、不适、发红）是由多形核白细胞（PMN）大量招募到阴道上皮所引发的。使用阴道炎小鼠模型，S100 报警素被确定为这一标志性 PMN 反应中的关键信号分子。然而，最近的研究使用 S100 警报素产生缺陷的小鼠，当受到白色念珠菌的攻击时，仍然会出现剧烈的 PMN 募集反应。因此，关于念珠菌阴道炎期间中性粒细胞被募集到阴道上皮的替代和/或补充机制，在知识上仍然存在根本差距。 在上皮表面引发炎症的一个可能过程是通过激活炎症小体，炎症小体是一组发出信号并启动先天免疫反应的胞质细胞受体。因此，该提案的目的是 1) 证明 Nlrp3 炎症小体是先天炎症反应的关键早期介质，2) 识别导致阴道上皮炎症小体激活的特定菌丝相关白色念珠菌毒力因子。在强有力的初步数据的指导下，这些目标将检验我们的中心假设，即阴道炎期间白色念珠菌酵母菌向菌丝转变诱导的炎性小体的激活会加剧阴道上皮中的中性粒细胞流入和促炎细胞因子信号传导。所有目标都将利用成熟的阴道炎小鼠模型来检验我们的假设。在第一个目标下，将利用炎症体信号级联中离散靶点的遗传或药理学阻断来阐明导致念珠菌阴道炎的信号通路。将通过测量炎症标志物（PMN、Interleukin-1b、S100 警报素）来评估是否成功破坏炎症小体依赖性信号传导。在第二个目标中，缺乏特定菌丝相关毒力因子（分泌的蛋白酶或粘附素）的具有形态发生能力的白色念珠菌菌株或锁定在酵母形式但过度表达这些关键已识别菌丝毒力因子的菌株将被用来评估其所需的在体内产生强大的先天免疫反应的作用。这种方法具有创新性，因为它检查了与念珠菌阴道炎相关的免疫反应所需的宿主和真菌过程，并试图识别阴道粘膜中的炎性体功能。这项拟议的工作意义重大，因为它将最终有助于识别新的信号传导途径，这些信号传导途径可以作为此类感染的新治疗靶点。 这项工作还将为候选人获得独立的学术研究项目奠定坚实的基础。该候选人的近期职业目标是成功从非终身教授职位的初级教员过渡到终身职位助理教授职位，研究宿主先天免疫反应和与真菌性阴道炎相关的分子机制。他的长期职业目标是获得终身研究教职 领导和维持一个资金充足的转化研究团队，研究粘膜界面真菌病原体的先天免疫机制。为了实现这些职业目标，除了进行上述研究之外，候选人还将组建并维持一个支持性的指导/咨询团队，这将有助于成功完成研究，磨练资助技能的发展，从而获得有效的资助机会，并加快高影响力数据的发布。因此，这些职业目标实现了 NIAID 的使命是寻找新的治疗方法来对抗真菌病，并支持初级研究人员在 K22 奖励机制下实现职业转型，成为独立教职人员。

项目成果

Candidalysin Drives Epithelial Signaling, Neutrophil Recruitment, and Immunopathology at the Vaginal Mucosa.

念珠菌溶酶驱动阴道粘膜的上皮信号传导、中性粒细胞募集和免疫病理学。

• 发表时间: 2018
• 影响因子: 3.1
• 作者: Richardson, Jonathan P;Willems, Hubertine M E;Moyes, David L;Shoaie, Saeed;Barker, Katherine S;Tan, Shir Lynn;Palmer, Glen E;Hube, Bernhard;Naglik, Julian R;Peters, Brian M
• 通讯作者: Peters, Brian M

Polymicrobial Biofilm Studies: From Basic Science to Biofilm Control.

多种微生物生物膜研究：从基础科学到生物膜控制。

• 发表时间: 2016-03
• 作者: Willems, Hubertine Me;Xu, Zhenbo;Peters, Brian M
• 通讯作者: Peters, Brian M