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：
10407531
负责人：
Danielle A Garsin
金额：
$ 46.53万
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/10407531
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.

项目概要/摘要口咽念珠菌病 (OPC) 是患者群体口腔发病的一个主要来源包括免疫抑制者（癌症和艾滋病患者）和服用广谱抗生素的人。我们最近发现了一种由粪肠球菌产生的肽，可以抑制菌丝形态发生， OPC 的主要病原体白色念珠菌形成生物膜。本申请中提出的研究复制旨在了解这种肽 EntV 的成熟形式是如何产生的，它如何靶向白色念珠菌罐头菌丝形态发生，以及它对OPC的保护程度，从而填补了我们的一个关键空白知识并为治疗学的发展提供新的途径。本次重组的长远目标是研究的目的是开发治疗和预防口咽念珠菌病的新策略。的目标该应用旨在确定 EntV 的产生、作用方式和治疗潜力。中央假设是分泌和加工的 EntV 作用于控制菌丝发育的信号通路白色念珠菌，可预防念珠菌病。这项研究的基本原理是确定 EntV 抑制白色念珠菌侵袭性形成的机制将带来新的治疗方法策略。通过追求三个目标，本申请的目标将得以实现。目标 #1 将阐明如何生成 EntV 的活性形式。我们将使用遗传和生化方法来检验该假设裂解需要蛋白酶 GelE 和/或 SprE，并且二硫键的形成是由 DsbA 同系物。在目标#2中，白色念珠菌的基因/途径被粪肠球菌靶向抑制菌丝形态发生将使用细胞生物学和遗传学方法来确定。根据初步数据，工作假设是 EntV 直接与 C 上几个已知的离散子域之一相互作用。白色念珠菌细胞表面破坏细胞与细胞和细胞与基质的粘附，这两者对于生物膜都是必需的形成。 EntV 和相关肽在预防白色念珠菌感染方面的功效将在目标#3。 OPC 和胃肠道激素的组织培养细胞（巨噬细胞和口腔上皮细胞）和小鼠模型将利用睾丸定植。我们假设 EntV 将防止白色念珠菌感染，并且可以- 纳摩尔范围内的糖尿病。这一贡献的意义在于了解细菌如何肽抑制白色念珠菌菌丝形态发生，可能为治疗设计开辟新途径。将菌丝形态发生作为可能开发新疗法的一个领域是创新的，因为目前的抗真菌药物针对细胞膜的完整性或生长。这样做的额外好处研究包括细菌肽生成、真菌形态类型转换等领域的新知识界间微生物相互作用。