Identification of early phase C. albicans biofilm proteins

早期白色念珠菌生物膜蛋白的鉴定

• 批准号: 8063533
• 负责人: Mahmoud A Ghannoum
• 金额: $ 35.08万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-05-15 至 2012-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8063533
• 关键词: Adherence; Alcohol dehydrogenase; Animal Experimentation; Animal Model; Architecture; Azole resistance; Biochemical; Biological Assay; Biology; Bromodeoxyuridine; Canada; Candida; Candida albicans; Candidiasis; Cell Death; Cell Wall; Cell membrane; Cells; Characteristics; Clinical; Collaborations; Collagen Type IV; Complication; Data; Denture Stomatitis; Dentures; Development; Devices; Diagnosis; Diagnostic; Early identification; Ensure; Exhibits; Fluorescence Microscopy; Fungal Drug Resistance; Gene Proteins; Genes; Growth; HIV; Histopathology; Human Engineering; Hyphae; Illinois; In Vitro; Infection; Invaded; Laboratories; Laser Scanning Confocal Microscopy; Link; Manuscripts; Mass Spectrum Analysis; Measurement; Measures; Metabolic; Metabolic Pathway; Methods; Microbial Biofilms; Microscopy; Modeling; Molecular; Monitor; Mouth Diseases; Mucous Membrane; Mus; Opportunistic Infections; Oral; Oral candidiasis; Oral cavity; Oral mucous membrane structure; Parents; Pathogenesis; Pathway Analysis; Pathway interactions; Patients; Phase; Phenotype; Play; Prevention; Principal Investigator; Production; Proteins; Proteomics; Publications; Research; Research Personnel; Resistance profile; Role; Scanning Electron Microscopy; Shipping; Ships; Stimulus; Surface; Techniques; Testing; Therapeutic; Thick; Tissues; Tongue; Universities; Weight; Yeasts; base; candida biofilm; gel electrophoresis; genetic manipulation; in vitro Model; in vivo; in vivo Model; insight; laminin-5; liquid chromatography mass spectrometry; microorganism; mutant; oral infection; oropharyngeal thrush; programs; protein expression; research clinical testing; research study; tool; two-dimensional

DESCRIPTION (provided by applicant): Oropharyngeal candidiasis (OPC, thrush) is the term given to opportunistic oral infection caused by the yeast Candida, and is the most common mycotic complication associated with human immunodeficiency virus (HlV)-infected patients. The ability of Candida isolates to form biofilm on devices and host tissue surfaces is believed to be intricately linked with OPC infections and denture stomatitis. Therefore, understanding the role of biofilms in pathogenesis and host-Cand/cfa interactions is critical. In this application, to gain insight into these interactions, we will use a proteomics-based approach to identify specific proteins that are central to the biofilm forming ability of C. albicans and determine their role in interactions between fungal biofilm and host tissues. We have previously established an in vitro model of C. albicans denture biofilm (Publication 1, Appendix). Using this model, we: 1) identified the developmental phases of candidal biofilm formation (Publication 1, Appendix), 2) investigated the antifungal resistance profile of C. albicans biofilms at different growth phases (Publication 2, Appendix), 3) investigated the multifactorial mechanisms of azole resistance of early and mature biofilms formed by C. albicans (Publication 3, Appendix), 4) used a proteomic approach to identify potential target proteins that are differentially expressed in early phase biofilms (see Publication 4, Appendix), 5) used molecular and biochemical methods to show that one of the identified proteins (alcohol dehydrogenase, Adhlp) is a negative regulator of Candida biofilm (Publication 4, Appendix), and 6) moved our studies closer to the clinical setting by using an engineered human oral mucosa (EHOM), developed recently by Dr. Rouabhia (Co-investigator on this application). This collaborative research between the applicant and Dr. Rouabhia showed that Adhlp plays an important role in both Candida biofilm formation and invasion of host mucosal tissues (Manuscript submitted, Appendix). The overall hypothesis of this application is that C. albicans express specific proteins that are essential for biofilm formation and play critical roles in Candida-host tissue interactions. We will test our hypothesis using the following Specific Aims: Aim 1. Identify early phase biofilm-specific proteins expressed by C. albicans. Aim 2. Determine whether the identified proteins are critical to the ability of C. albicans to form biofilms in vitro by disrupting genes encoding them. Aim 3. Use an in v/Vo-like Engineered Human Oral Mucosa (EHOM) Model to determine whether the identified proteins are essential for Candida biofilm formation and host tissue damage. Aim 4. Validate the in vitro and EHOM results in vivo using a murine oral model of Candida biofilms. Experiments described in this application will provide insight into the biology of OPC-associated C. albicans biofilms and may suggest potential therapeutic and diagnostic targets for the prevention, treatment, and diagnosis of oral Candida infections.

描述（由申请人提供）：口咽念珠菌病（OPC，鹅口疮）是由酵母念珠菌引起的机会性口腔感染的术语，是与人类免疫缺陷病毒（HLV）感染患者有关的最常见的霉菌并发症。据信，念珠菌分离株在设备和宿主组织表面上形成生物膜的能力与OPC感染和牙齿口齿炎相关。因此，了解生物膜在发病机理和宿主/CFA相互作用中的作用至关重要。在此应用中，为了深入了解这些相互作用，我们将使用基于蛋白质组学的方法来识别特定的蛋白质，这些蛋白质对白色念珠菌的生物膜形成能力至关重要，并确定它们在真菌生物膜和宿主组织之间相互作用中的作用。我们以前已经建立了白色念珠菌义齿生物膜的体外模型（出版物1，附录）。 Using this model, we: 1) identified the developmental phases of candidal biofilm formation (Publication 1, Appendix), 2) investigated the antifungal resistance profile of C. albicans biofilms at different growth phases (Publication 2, Appendix), 3) investigated the multifactorial mechanisms of azole resistance of early and mature biofilms formed by C. albicans (Publication 3, Appendix), 4) used一种蛋白质组学方法，以鉴定在早期生物膜中差异表达的潜在靶蛋白（请参见出版物4，附录），5）使用分子和生化方法表明，通过使用candida Biofilm（abublic oferation 4，abublitix Insentix），使用分子和生化方法表明，鉴定出的蛋白质之一（酒精脱氢酶，ADHLP，ADHLP）是一种负面的调节器，以及6）。口服粘膜（EHOM），由Rouabhia博士（该应用程序的共同研究器）最近开发。申请人和鲁比亚博士之间的这项合作研究表明，ADHLP在念珠菌生物膜形成和入侵宿主粘膜组织（手稿提交，附录）中起着重要作用。该应用的总体假设是白色念珠菌表达对生物膜形成至关重要的特定蛋白质，并在念珠菌宿主组织相互作用中起关键作用。我们将使用以下特定目的检验我们的假设：目标1。确定白色念珠菌表达的早期生物膜特异性蛋白。 AIM 2。确定鉴定的蛋白是否对白色念珠菌在体外形成生物膜的能力是否至关重要，通过破坏编码它们的基因。 AIM 3。使用类似V/VO的工程人口腔粘膜（EHOM）模型来确定所鉴定的蛋白是否对于念珠菌生物膜形成和宿主组织损伤至关重要。目标4。使用念珠菌生物膜的鼠口服模型验证体外和EHOM会导致体内。本应用中描述的实验将洞悉与OPC相关的白色念珠菌生物膜的生物学，并可能提出预防，治疗和诊断口服念珠菌感染的潜在治疗和诊断靶标。

项目成果

Identification of gentian violet concentration that does not stain oral mucosa, possesses anti-candidal activity and is well tolerated.

• DOI: 10.1007/s10096-010-1131-8
• 发表时间: 2011-05
• 期刊: EUROPEAN JOURNAL OF CLINICAL MICROBIOLOGY & INFECTIOUS DISEASES
• 影响因子: 4.5
• 作者: Jurevic, R. J.;Traboulsi, R. S.;Mukherjee, P. K.;Salata, R. A.;Ghannoum, M. A.
• 通讯作者: Ghannoum, M. A.