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.

项目摘要/摘要 我有理想的经验水平和机构环境，可以最大限度地从职业中受益 发展奖。我的研究经历始于本科，当时我是一项研究的一部分 使用核磁共振光谱学解决了RNA结构问题的团队。我跟随这个项目进行了一项癌症研究 培训奖和在美国国立卫生研究院的密集研究经验。在医疗期间专注于临床护理之后 在学校和儿科住院医生的培训中，我回到了基础科学研究领域，成为儿科传染病 伙计。我获得了享有盛誉的儿科传染病学会奖学金，该奖项支持 我的研究员研究和作为初级教员进行的第四年指导研究。在Sarah Gaffen博士的指导下，我 致力于研究粘膜念珠菌感染的先天和适应性宿主反应，并发起了 口咽念珠菌病中性粒细胞与IL-17关系的研究我还调查了 IL-17在播散性念珠菌病真菌负荷和疾病病理中的作用总而言之，我是一个作家 关于9篇关于念珠菌病和/或IL-17的手稿，其中4篇是第一作者。我的研究项目 自从8月份开始在威斯康星医学院(MCW)担任助理教授以来，这种情况一直在继续， 2014年。MCW的环境为拟议的项目提供了一个出色的场地。尤其是， 充满活力的校园免疫学小组鼓励合作和互动。尽管我还在继续 与Gaffen博士合作，我已经为我当前的项目与专家建立了相关的指导关系 趋化因子结构生物学和粘膜免疫学(博士Brian Volkman和Mitchell Grayson， )。我的实验室参加免疫学、微生物学和传染病研讨会。我 拥有良好的实验室空间，为动物研究提供充足的支持，并可获得所有设备和核心资源 完成这个项目所需要的，包括流式细胞仪、组织学服务和生物统计学 通过妇幼保健会儿童研究所提供支持。 我的整个职业目标是成为一名独立的、由NIH资助的内科科学家，专注于翻译 与真菌感染有关的粘膜免疫学研究。我是由我的顾问指导的 委员会，由当地的粘膜免疫学、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，以 增加我对负责任的研究、稿件和资助评审以及研究的知识- 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 免疫学和生物统计学。在获奖期的后两年，我打算撰写并提交一份 NIH R01申请成功。在正在进行的指导和评估方面，我将会见我的每一位 独立的导师每两周一次，与我的导师委员会每季度一次，以确保两项目标的实现 研究目标和自主成长。 在目前的方案中，我们旨在研究趋化因子CCL28在口腔粘膜免疫中的作用 死于真菌感染。口咽念珠菌病(OPC)是一种常见且严重的疾病。 免疫功能低下的个体和发展为高死亡率播散性疾病的风险标志 屏障和免疫缺陷患者中的念珠菌病。CCL28是一种具有体外抗菌活性的CC趋化因子 多肽(AMP)活性，淋巴细胞和嗜酸性粒细胞趋化活性，以及粘膜定位。我们的 研究CCL28在粘膜宿主抗感染防御中的方法和原理是基于初步的 OPC中抗念珠菌基序和基因表达的数据。CCL28的S活动可能填补中国的空白 目前的OPC宿主防御模型包括促炎细胞因子、细胞募集和 安培。我们建议使用已取消AMP的构建变体来研究CCL28的功能 趋化活性。我们假设CCL28在粘膜界面上直接作为一种 AMP，并间接通过诱导中性粒细胞趋化物质。AIMS采取了创新的方法 应用结构生物学手段研究小分子免疫蛋白的作用机制 限制感染。在目标1中，我们将描述趋化因子和细胞因子表达的调控。 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 CCL28各功能在OPC小鼠模型中的活性和效力。使用结构函数数据来 最大化CCL28在OPC中的S效力，同时最小化病理炎症活动将使我们能够利用 CCL28的生物学特性，并扩大了对免疫机制的理解。总体而言，这一K奖将 允许我在与粘膜宿主防御相关的分子的结构-功能分析中获得独立性。我 将能够应用这些技能来获得未来的R01资金，作为主持人的独立调查员 对真菌感染的免疫反应，包括AMP和趋化因子，以提高对风险的认识 感染因子，并开发新的治疗方法。