Anti-Candida activity of CCL28 in oropharyngeal candidiasis

CCL28 在口咽念珠菌病中的抗念珠菌活性

• 批准号: 9902401
• 负责人: Anna Huppler
• 金额: $ 16.13万
• 依托单位: MEDICAL COLLEGE OF WISCONSIN
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-05-01 至 2022-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9902401
• 关键词: Achievement; Advanced Development; Animals; Antifungal Agents; Asthma; Authorship; Autopsy; Award; Basic Science; Biological; Biological Assay; Biometry; CC chemokine receptor 3; CSF3 gene; CXCL1 gene; CXCL5 gene; Candida; Candida albicans; Candidiasis; Categories; Cell-Free System; Cells; Cellular Immunology; Charge; Chemotactic Factors; Chemotaxis; Chemotaxis Induction; Child; Childhood; Collaborations; Communicable Diseases; Data; Development; Disease; Disseminated candidiasis; Ensure; Environment; Epithelial Cells; Equipment; Evaluation; Faculty; Fellowship; Funding; Future; GPR2 gene; Gene Expression; Goals; Grant; Grant Review; Growth; Histological Techniques; Histology; Host Defense; Immune; Immune response; Immunocompromised Host; Immunodeficient Mouse; Immunology; In Vitro; Incidence; Individual; Infection; Inflammation; Inflammatory; Interleukin-17; Interleukin-6; Interleukin-8B Receptor; Investigation; K-Series Research Career Programs; Knowledge; Laboratories; Length; Leukocytes; Lymphocyte; Lymphocyte Subset; Manuscripts; Mediating; Mentors; Mentorship; Microbiology; Modeling; Mucosal Immunity; Mucous Membrane; Mus; Mutation; Mycoses; NMR Spectroscopy; Neutrophil Infiltration; Oral; Oral cavity; Oral mucous membrane structure; Oropharyngeal; Pathogenesis; Pathologic; Pathology; Patients; Physicians; Predisposition; Property; Proteins; RNA; Recombinants; Regulation; Research; Research Institute; Research Personnel; Research Project Grants; Research Training; Residencies; Resistance; Resources; Respiratory Mucosa; Risk Factors; Risk Marker; Rodent; Role; Salivary; Salivary Glands; Scientist; Services; Societies; Structure; Testing; Therapeutic; Topical application; Training; Translational Research; United States National Institutes of Health; Up-Regulation; Variant; Viral; Wild Type Mouse; Wisconsin; Work; Writing; anticancer research; antimicrobial; antimicrobial peptide; base; beta-Chemokines; beta-Defensins; burden of illness; career; chemokine; chemokine receptor; clinical care; cytokine; eosinophil; experience; histatin 5; immunoregulation; improved; in vivo; infection burden; innovation; insight; interleukin-22; medical schools; mortality; mouse model; neutrophil; novel therapeutics; oropharyngeal thrush; pathogen; professor; protein folding; receptor binding; recruit; response; responsible research conduct; side effect; skills; structural biology; therapeutic candidate; tissue culture; tool; undergraduate student

PROJECT SUMMARY/ABSTRACT I have the ideal level of experience and institutional environment to maximally benefit from a Career Development Award. My research experiences started as an undergraduate when I was part of a research team who solved an RNA structure using NMR spectroscopy. I followed this project with a Cancer Research Training Award and an intensive research experience at the NIH. After focusing on clinical care during medical school and pediatric residency training, I returned to basic science research as a pediatric infectious diseases fellow. I was granted a prestigious Pediatric Infectious Diseases Society Fellowship Award which supported my fellowship research and a 4th year of mentored research as junior faculty. Mentored by Dr. Sarah Gaffen, I contributed to studies on the innate and adaptive host responses to mucosal Candida infections and initiated work on the relationship of neutrophils and IL-17 in oropharyngeal candidiasis. I also investigated the contribution of IL-17 to fungal burden and disease pathology in disseminated candidiasis. In all, I am an author on 9 manuscripts relating to candidiasis and/or IL-17 of which 4 are first authorships. My research projects have continued since starting as an assistant professor at the Medical College of Wisconsin (MCW) in August, 2014. The environment at MCW provides an outstanding venue for the proposed project. In particular, the vibrant campus Immunology group encourages collaboration and interaction. Although I continue to collaborate with Dr. Gaffen, I have established relevant mentorship for my current project with experts in chemokine structural biology and mucosal immunology (Drs. Brian Volkman and Mitchell Grayson, respectively). My laboratory participates in immunology, microbiology, and infectious diseases seminars. I have excellent lab space, ample support for animal studies, and access to all equipment and core resources required for the completion of this project including flow cytometers, histology services, and biostatistics support through the MCW Children’s Research Institute. My overall career goal is to be an independent, NIH-funded physician scientist focused on translational research in mucosal immunology as it relates to fungal infections. I am guided by my advisory Mentorship Committee, composed of local experts in mucosal immunology, structural biology, antimicrobial peptides, cellular immunology, chemokines, and pathogenesis (mentors Volkman and Grayson, as well as Drs. Nita Salzman, Bonnie Dittel, Michael Dwinell, and Jenifer Coburn). I plan training and mentorship activities to augment my knowledge of the responsible conduct of research, manuscript and grant reviews, and research- related topics including tissue culture, chemotaxis, rodent necropsy, histological techniques, mucosal immunology, and biostatistics. During the latter two years of the award period I intend to write and submit a successful NIH R01 application. In terms of ongoing guidance and evaluation, I will meet with each of my mentors independently biweekly and with my Mentorship Committee quarterly to ensure both achievement of research goals and growth towards independence. In the current proposal, we aim to investigate the function of the chemokine CCL28 in oral mucosal immunity from fungal infection. Oropharyngeal candidiasis (OPC) is a frequent and serious problem for immunocompromised individuals and a marker of risk for the development of high-mortality disseminated candidiasis in patients with barrier and immune deficits. CCL28 is a CC chemokine with in vitro antimicrobial peptide (AMP) activity, chemotactic activity for lymphocytes and eosinophils, and mucosal localization. Our approach and rationale for investigating CCL28 in mucosal host defense from infection is based on preliminary data on the anti-Candida motifs and gene expression in OPC. CCL28’s activities potentially fill gaps in the current model of OPC host defense which includes pro-inflammatory cytokines, cellular recruitment, and AMPs. We propose to study the function of CCL28 using constructed variants with abolished AMP or chemotactic activity. We hypothesize that CCL28 controls Candida at the mucosal interface directly as an AMP and indirectly through induction of neutrophil chemoattractants. The aims take an innovative approach of applying structural biology tools to determine the mechanisms of small immune proteins and their role in limiting infection. In Aim 1, we will characterize the regulation of chemokine and cytokine expression by CCL28 and determine the active motif for each distinct function. From the studies in Aim 2, we will determine the activity and potency of each CCL28 function in the mouse model of OPC. Using structure-function data to maximize CCL28’s potency in OPC while minimizing pathologic inflammatory activity will allow us to harness the biologic properties of CCL28 and expand understanding of immune mechanisms. Overall, this K award will allow me to gain independence in structure-function analysis of molecules relevant to mucosal host defense. I will be able to apply these skills to obtain future R01 funding as an independent investigator on the host immune response to fungal infections, including AMPs and chemokines, in order to improve recognition of risk factors for infection and develop new therapeutics.

项目概要/摘要 我拥有理想水平的经验和制度环境，可以从职业生涯中最大限度地受益 发展奖 我的研究经历是从本科生开始的，当时我参与了一项研究。 团队使用 NMR 光谱法解决了 RNA 结构 我通过癌症研究跟踪了这个项目。 在医学期间专注于临床护理后，获得了培训奖并在 NIH 获得了深入的研究经验。 学校和儿科住院医师培训，我回到儿科传染病基础科学研究 我获得了著名的儿科传染病协会奖学金奖，该奖支持 我的奖学金研究和作为初级教师的第四年指导研究，由莎拉·加芬博士指导。 有助于研究宿主对粘膜念珠菌感染的先天性和适应性反应，并发起 我还研究了中性粒细胞和 IL-17 在口咽念珠菌病中的关系。 IL-17 对播散性念珠菌病真菌负荷和疾病病理学的贡献 总之，我是一名作者。 9 篇有关念珠菌病和/或 IL-17 的手稿，其中 4 篇是我的研究项目。 自八月份开始在威斯康星医学院 (MCW) 担任助理教授以来， 2014. MCW 的环境为拟议项目提供了绝佳的场地。 充满活力的校园免疫学小组鼓励合作和互动，尽管我继续这样做。 与加芬博士合作，我与以下领域的专家为我当前的项目建立了相关的指导 趋化因子结构生物学和粘膜免疫学（Brian Volkman 博士和 Mitchell Grayson 博士， 我的实验室参加了免疫学、微生物学和传染病研讨会。 拥有优良的实验室空间、对动物研究的充足支持以及所有设备和核心资源的使用权 完成该项目所需的资源，包括流式细胞仪、组织学服务和生物统计学 通过 MCW 儿童研究所提供支持。 我的总体职业目标是成为一名独立的、由 NIH 资助的医师科学家，专注于转化 我在我的顾问指导下进行了与真菌感染相关的粘膜免疫学研究。 委员会由当地粘膜免疫学、结构生物学、抗菌肽、 细胞免疫学、趋化因子和发病机制（导师 Volkman 和 Grayson，以及 Nita 博士） Salzman、Bonnie Dittel、Michael Dwinell 和 Jenifer Coburn）我计划培训和指导活动 增强我对负责任的研究行为、手稿和资助审查以及研究的了解- 相关主题包括组织培养、趋化性、啮齿动物尸检、组织学技术、粘膜 在奖励期的后两年，我打算撰写并提交一份关于免疫学和生物统计学的文章。 成功的 NIH R01 申请 在持续的指导和评估方面，我会与我的每个人见面。 每两周独立辅导一次，每季度与我的辅导委员会一起辅导，以确保实现 研究目标和走向独立的成长。 在当前的提案中，我们旨在研究趋化因子CCL28在口腔粘膜免疫中的功能 口咽念珠菌病 (OPC) 是一种常见且严重的问题。 免疫功能低下的个体和传播高死亡率的风险标志 CCL28 是一种具有体外抗菌作用的 CC 趋化因子。 肽 (AMP) 活性、淋巴细胞和嗜酸性粒细胞的趋化活性以及粘膜定位。 研究 CCL28 在粘膜宿主感染防御中的方法和基本原理基于初步 OPC 中抗念珠菌基序和基因表达的数据可能填补了该领域的空白。 目前 OPC 宿主防御模型包括促炎细胞因子、细胞募集和 我们建议使用已废除 AMP 的构建变体来研究 CCL28 的功能。 我们勇敢地说，CCL28 可以直接控制粘膜界面的念珠菌。 AMP 并通过诱导中性粒细胞化学引诱剂间接实现。 用于确定小免疫蛋白机制及其作用的结构生物学工具 在目标 1 中，我们将通过以下方式来表征趋化因子和细胞因子表达的调节。 CCL28 并确定每个不同功能的活性基序 根据目标 2 中的研究，我们将确定。 使用结构功能数据来分析 OPC 小鼠模型中每个 CCL28 功能的活性和效力。 最大限度地发挥 CCL28 在减少 OPC 方面的功效，同时病理性炎症活动将使我们能够利用 总体而言，该 K 奖将研究 CCL28 的生物学特性并扩大对免疫机制的了解。 让我能够独立进行与粘膜宿主防御相关的分子的结构功能分析。 将能够应用这些技能作为主机的独立调查员获得未来的 R01 资金 对真菌感染的免疫反应，包括 AMP 和趋化因子，以提高对风险的识别 感染因素和开发新疗法。