Quantitative genetic approaches to Candida albicans oropharyngeal pathogenesis

白色念珠菌口咽发病机制的定量遗传学方法

• 批准号: 10007581
• 负责人: Robert John Fillinger
• 金额: $ 3.65万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-01-01 至 2022-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10007581
• 关键词: Address; Biology; Body Temperature; Candida; Candida albicans; Candidate Disease Gene; Cell Line; Clinical; Collection; Communication; Communities; Complex; Computers; Deglutition; Detection; Discipline; Disease; Dissection; Disseminated candidiasis; Environment; Epithelial; Epithelial Cells; Filament; Gastrointestinal tract structure; Genes; Genetic; Genetic Determinism; Genetic Recombination; Genetic Screening; Genetic Variation; Genome; Genotype; Goals; HIV; Human; Image; Immune; Immune response; Immune system; Immunocompromised Host; In Vitro; Individual; Infection; Informatics; International; Investigation; Lead; Life; Measurable; Meiosis; Methods; Modeling; Mouth Diseases; Mucous Membrane; Oral; Oral candidiasis; Oral cavity; Oropharyngeal; Outcome; Partner in relationship; Pathogenesis; Pathogenicity; Pathology; Patients; Phenotype; Ploidies; Positioning Attribute; Procedures; Process; Production; Quantitative Genetics; Quantitative Trait Loci; Recombinants; Redness; Regulation; Regulatory Pathway; Research; Risk; Role; Science; Scientist; Signal Transduction; Site; Skin; Surface; Swelling; Systemic disease; Systemic infection; Techniques; Testing; Time; Transplant Recipients; Urinary tract; Variant; Virulence; Virulence Factors; Visual; Work; Writing; Yeasts; cell injury; cell mediated lymphocytolysis test; clinically relevant; disease phenotype; experimental study; gene discovery; genetic approach; genomic locus; graduate student; immune activation; immune clearance; immunological status; mutant; neonate; new therapeutic target; novel; opportunistic pathogen; oral cavity epithelium; oropharyngeal thrush; pathogenic fungus; reference genome; skills; species difference; trait

Abstract: Candida albicans is a polymorphic commensal yeast that also manifests as a prevalent opportunistic fungal pathogen. C. albicans infections manifest as oropharyngeal candidiasis (OPC), or thrush, in immunocompromised individuals. Here, we deploy quantitative trait loci (QTL) analysis to identify genes responsible for different forms of oral disease and to dissect regulatory networks governing the pathogenic process of filamentation in C. albicans. These experiments will develop quantitative genetic approaches for all parasexual Candida species. C. albicans has substantial phenotypic diversity among strains and is capable of residing stably in the oral cavity or inducing epithelial damage that leads to either clearance by the immune system or OPC. However, the genetic loci governing differences in these disease phenotypes between strains are not understood. To address this, two isolates 529L and SC5314, a stable colonizer and damager, respectively, were mated to generate parasexual progeny. Progeny genomes were sequenced at marker positions covering ~1% of the genome. High rates of recombination during parasex facilitated mapping of 19 QTL regions associated with epithelial damage phenotypes using a chromium release assay. Aim 1 will test specific loci from within those QTLs to identify genetic determinants of damage and clearance by the immune system or non-damaging persistence that differentiate strains. Furthermore, QTL analysis of filamentation will be performed, as hyphal formation and host cell damage were separable phenotypes in some progeny. C. albicans uses filamentation to disrupt epithelial surfaces and progress into a disseminated systemic infection. Current models of filamentation do not account for the substantial differences in filamentation among C. albicans isolates. To identify species-wide regulators of filamentation, we will employ QTL techniques using 4 strains with distinct filamentation phenotypes in Aim 2. QTLs from all pairwise matings will uncover genes regulating differences in filamentation phenotypes when grown on three different induction media at 30oC and 37oC to mimic the oral cavity and internal body temperature of the human host. Filamentation phenotyping will be scored with a novel high-throughput visual analysis procedure. Thus, establishment of quantitative genetic approaches in a parasexual species will greatly enhance identification of C. albicans pathogenesis regulators. My goal as a graduate student is to become a scientist whose research brings together multiple scientific disciplines. While pursuing these aims, I will develop my skills at the bench and at the computer to bridge the gap between informatics and biology, an increasingly important role in science. Throughout this project, I will develop my communication skills by presenting this work to scientific audiences locally and internationally, collaborating with other scientists, and writing my work for broader dissemination to the scientific and lay communities.

抽象的： 白色念珠菌是一种多态性酵母，也表现为普遍的机会主义 真菌病原体。白色念珠菌感染表现为口咽念珠菌病（OPC）或鹅口疮， 免疫功能低下的个体。在这里，我们部署定量性状基因座（QTL）分析以识别基因 负责不同形式的口腔疾病，并剖析控制致病性的调节网络 白色念珠菌的细丝过程。这些实验将为所有人开发定量遗传方法 寄生虫念珠菌。 白色念珠菌在菌株之间具有实质性的表型多样性，并且能够稳定地居住在 口腔或诱导上皮损伤，导致免疫系统或OPC清除。然而， 尚不清楚菌株之间这些疾病表型的遗传基因座差异。到 解决这个问题，分别将两个分离株和SC5314分别为稳定的菌落和Damager 产生寄生虫后代。对后代基因组在覆盖约1％的标记位置进行测序 基因组。帕拉塞克斯期间的高重组速率促进了与19个QTL区域的映射 使用铬释放测定法上皮损伤表型。 AIM 1将从其中测试特定基因座 QTL识别免疫系统或不损害的损害和清除率的遗传决定因素 区分菌株的持久性。此外，将作为菌丝进行QTL丝分析 在某些后代中，形成和宿主细胞损伤是可分离的表型。 白色念珠菌使用细丝破坏上皮表面并发展为传播的系统性 感染。当前的细丝模型并未解释丝之间的实质性差异 白色念珠菌分离株。为了识别细丝的整个物种调节剂，我们将使用QTL技术 在AIM 2中具有独特细丝表型的4个菌株。来自所有成对分子的QTL将发现基因 在30oC和 37oC模仿人类宿主的口腔和内体温度。细丝表型将 通过新型的高通量视觉分析程序进行评分。因此，建立定量遗传 偏二种物种中的方法将大大增强白色念珠菌发病机理调节剂的鉴定。 我作为研究生的目标是成为一名科学家，其研究汇集了多个科学 学科。在追求这些目标的同时，我将在板凳和计算机上发展自己的技能，以弥合 信息学和生物学之间的差距，这是科学中日益重要的作用。在整个项目中，我将 通过向本地和国际科学观众介绍这项工作来发展我的沟通技巧 与其他科学家合作，写我的工作，以更广泛地传播科学并进行。 社区。