Inhibition of Candida virulence and biofilm formation by a bacterial peptide

细菌肽抑制念珠菌毒力和生物膜形成

基本信息

批准号：
10621569
负责人：
Danielle A Garsin
金额：
$ 7.6万
依托单位：
UNIVERSITY OF TEXAS HLTH SCI CTR HOUSTON
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-07-01 至 2024-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10621569
关键词：
Acquired Immunodeficiency Syndrome Actins Adherence Adhesions Affect Amino Acids Animal Model Antibiotics Antifungal Agents Area Attenuated Bacteria Bacterial Adhesins Binding Biochemical Biogenesis Biological Biology Caenorhabditis elegans Candida Candida albicans Candidiasis Cell Surface Proteins Cell Wall Cell surface Cells Chemicals Data Development Disease model Enterococcus faecalis Epithelial Epithelial Cells Felis catus Generations Genes Genetic Glycoproteins Goals Growth HIV Seropositivity Head and neck structure Homologous Gene Human Microbiome Immunocompromised Host In Vitro Infant Infection Inflammation Investigation Knowledge Label Malignant Neoplasms Medical Membrane Microdomains Microbial Biofilms Modeling Morbidity - disease rate Morphogenesis Mus Oral Oral candidiasis Oral cavity Outcome Pathway interactions Patients Peptide Hydrolases Peptides Post-Translational Protein Processing Process Proteins Public Health Radiation Research Risk Role Signal Pathway Site Source Steroids Surface Testing Therapeutic Vertebrates Virulence Work Yeasts base cell envelope cell growth chemotherapy design disulfide bond efficacy testing experimental study fungus gastrointestinal gene product genetic approach gut colonization immunosuppressed innovation macrophage microorganism interaction mouse model mutant nanomolar normal microbiota novel novel strategies novel therapeutics oral cavity epithelium oral infection oropharyngeal thrush prevent therapeutic development tissue/cell culture treatment strategy

项目摘要

PROJECT SUMMARY/ABSTRACT The medical condition oropharyngeal candidiasis (OPC) is a major source of oral morbidity in patient groups including the immunosuppressed (cancer and AIDS patients) and those taking broad-spectrum antibiotics. We recently identified a peptide produced by Enterococcus faecalis that inhibits the hyphal morphogenesis and biofilm formation of the primary causative agent of OPC, Candida albicans. The research proposed in this ap- plication seeks to understand how the mature form of this peptide, EntV, is generated, how it targets C. albi- cans hyphal morphogenesis, and the extent to which it protects against OPC, thereby filling a critical gap in our knowledge and providing new avenues for the development of therapeutics. The long-term goal of this re- search is to develop novel strategies for treating and preventing oropharyngeal candidiasis. The objective of this application is to determine the generation, mode of action and therapeutic potential of EntV. The central hypothesis is that secreted and processed EntV acts on signaling pathway(s) that controls hyphal development in C. albicans resulting in protection from candidiasis. The rationale for this research is that identification of the mechanism by which EntV inhibits the formation of the invasive form of C. albicans will lead to new treatment strategies. By pursing three aims, the objective of this application will be attained. Aim #1 will elucidate how the active form of EntV is generated. Using both genetic and biochemical approaches, we will test the hypothesis that the proteases GelE and/or SprE are required for cleavage, and disulfide bond formation is catalyzed by a DsbA homolog. In Aim #2, the genes/pathways of C. albicans that are targeted by E. faecalis to inhibit hyphal morphogenesis will be determined using cell biological and genetic approaches. Based on preliminary data, the working hypothesis is that EntV interacts directly with one of several known discrete subdomains on the C. albicans cell surface to disrupt cell-cell and cell-substrate adherence, both of which are essential for biofilm formation. The efficacy of EntV and related peptides in protecting against C. albicans infection will be tested in Aim #3. Tissue culture cells (macrophages and oral epithelial cells) and mouse models of OPC and gastroin- testinal colonization will be utilized. We postulate that EntV will protect against C. albicans infection and can- didiasis in the nanomolar range. The significance of this contribution will be the knowledge of how a bacterial peptide inhibits C. albicans hyphal morphogenesis, potentially opening new avenues for therapeutic design. Targeting hyphal morphogenesis as an area for the possible development of novel therapeutics is innovative, as current antifungals are directed against the integrity or growth of the cell envelope. Additional benefits of this research include new knowledge in the fields of bacterial peptide generation, fungal morphotype switching, and inter-kingdom microbial interactions.

项目摘要/摘要医学状况口咽念珠菌病（OPC）是患者组口腔发病率的主要来源包括免疫抑制（癌症和艾滋病患者）和服用广谱抗生素的患者。我们最近确定了粪肠球菌产生的肽，该肽抑制了菌丝形态发生和 OPC念珠菌白色念珠菌的主要因果剂的生物膜形成。这项研究提出的研究 Plication试图了解该肽的成熟形式是如何产生的，它如何靶向Albi-C. albi-able 罐装菌丝形态发生及其预防OPC的程度，从而填补了我们的临界空白知识并提供了用于发展治疗剂的新途径。这个重新的长期目标搜索是制定治疗和预防口咽念珠菌病的新型策略。目的该应用是为了确定ENTV的产生，作用方式和治疗潜力。中央假设是分泌和处理的ENTV对控制菌丝发育的信号通路的作用在白色念珠菌中，可保护念珠菌病。这项研究的理由是确定 ENTV抑制侵入性白色念珠菌形式的形成的机制将导致新处理策略。通过追求三个目标，将实现本申请的目标。 AIM＃1将阐明如何生成ENTV的活动形式。使用遗传和生化方法，我们将检验假设裂解需要蛋白酶Gele和/或SPRE，并且二硫键形成是由A催化的 DSBA同源物。在AIM＃2中，粪肠球菌靶向的白色念珠菌的基因/途径抑制菌丝形态发生将使用细胞生物学和遗传方法确定。基于初步数据工作假设是ENTV与C上的几个已知离散子域之一直接相互作用。白色念珠菌细胞表面破坏细胞细胞和细胞基底粘附，这两种都对生物膜必不可少形成。 ENTV和相关肽在保护白色念珠菌感染中的功效将在目标＃3。组织培养细胞（巨噬细胞和口腔上皮细胞）以及OPC和胃蛋白酶的小鼠模型将利用睾丸定殖。我们假设ENTV将预防白色念珠菌感染和能力纳摩尔范围内的介氧曲。这项贡献的意义将是了解细菌的知识肽抑制白色念珠菌菌丝形态发生，可能为治疗设计打开新的途径。将菌丝形态发生作为可能发展新疗法的领域是创新的，由于当前的抗真菌剂针对细胞包膜的完整性或生长。这对此的其他好处研究包括在细菌肽产生，真菌形型切换和昆明间微生物相互作用。