N6-methyladenosine in Candida white-opaque switching and oral infection

念珠菌白色不透明转换和口腔感染中的 N6-甲基腺苷

基本信息

批准号：
10370682
负责人：
Allison Marly Porman Swain
金额：
$ 10.72万
依托单位：
UNIVERSITY OF COLORADO DENVER
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-03-01 至 2024-02-29
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10370682
关键词：
Advisory Committees Affect Animal Model Antifungal Agents Binding Sites Bioinformatics Biology Cancer Patient Candida Candida albicans Carbon Carbon Dioxide Cell Culture Techniques Cells Clinical Complex Cues DNA Deposition Development Diploidy Environment Epigenetic Process Exposure to Filament Gene Expression Gene Expression Profiling Genes Genetic Transcription Genotype Goals Heat-Shock Response High temperature of physical object Human Immunocompromised Host Impairment Individual Infection Investigation Knowledge Leadership Life Cycle Stages Life Style Maps Mediating Meiosis Mentors Messenger RNA Metabolic Metabolism Methyltransferase Microbe Microbial Biofilms Mitosis Modification Molecular Mucous Membrane Mus Mutate Nucleotides Oral Oral cavity Organism Outcome Pain Partner in relationship Pathogenicity Persons Pharmacotherapy Phase Phenotype Play Population Post-Transcriptional Regulation Process Production Property RNA Radiation Radiation therapy Reader Regulation Reporting Research Research Personnel Ribosomal RNA Role Saccharomyces cerevisiae Sampling Signal Transduction Site Source Stress Systemic infection Temperature Tertiary Protein Structure Testing Training Programs Virulence Yeasts career cell growth cell type chromosome loss clinically relevant epitranscriptomics functional outcomes insight malignant mouth neoplasm member metabolomics microbiome analysis microbiota mouse model mutant opportunistic pathogen oral infection oral microbiome oropharyngeal thrush pathogen radiation effect response skills

项目摘要

Project Summary / Abstract Candida albicans white-opaque switching is an epigenetic process that regulates its mating ability, virulence, and colonization of different niches in the human host. One of these niches is the oral cavity, where Candida species can cause painful infections called oropharyngeal candidiasis (OPC). While there are known environmental cues and transcriptional regulators that control the white-opaque switch, a mechanistic understanding of how specific environments trigger switching to one state or another and how this contributes to colonization and infection is lacking. This proposal will investigate how the RNA modification N6- methyladenosine (m6A) acts to mediate white-opaque switching and mating in C. albicans. Preliminary studies from my collaborator and I have found that cells lacking IME4, the m6A methyltransferase, are impaired in temperature-induced opaque to white switching and have an increased mating efficiency. As someone with a background in Candida white-opaque switching and m6A biology, I am uniquely qualified to investigate the hypothesis that m6A regulation mediates environmental signals that induce phenotypic changes that mediate infection by C. albicans. Aim 1 is to identify targets of m6A modification in white-opaque switching by using eCLIP to map single nucleotide sites of m6A modification and binding sites of MRB1, the m6A reader, as well as gene expression analysis in wild-type and ime4 mutant white and opaque cells. Aim 2 will determine how the environment regulates m6A in switching and filamentation by generating gene expression and metabolomic profiles of cells under environmental conditions such as high temperature or CO2 that induce white-opaque switching or filamentation. Aim 3 will investigate the role of m6A in oral colonization of C. albicans by using cell culture and mouse models of OPC. White-opaque switching regulates oral colonization, and we hypothesize that IME4 plays a role in this process. In this aim, I will also study how radiation affects Candida cells lacking IME4 and how it affects colonization of the murine oral niche. In aim 4 I will investigate the function of m6A in interactions with the oral microbiome. Many of the m6A modified genes I have identified are involved in interspecies interactions, suggesting that IME4 and m6A regulate these interactions within the host. This research plan will be the first to combine the fields of epitranscriptomics and Candida biology to study how m6A contributes to the lifestyle and pathogenicity of a clinically relevant fungal species, with the potential to pave the way for new anti-fungal drug treatments for OPC. I have proposed a comprehensive training program for the K99 phase of this proposal including acquisition of new skills in bioinformatics, metabolomics, animal models of oral infection, microbiome analyses, guidance from an advisory committee of prominent mentors and collaborators, and development of key professional and leadership skills. Collectively, this plan will facilitate my transition to independence during the R00 phase and prepare me for a career as an independent researcher.

项目概要/摘要白色念珠菌白色不透明转换是一种表观遗传过程，可调节其交配能力、毒力、以及人类宿主不同生态位的殖民化。这些生态位之一是口腔，念珠菌在那里念珠菌属物种可引起称为口咽念珠菌病 (OPC) 的痛苦感染。虽然已知有控制白色不透明开关的环境线索和转录调节器，这是一种机制了解特定环境如何触发切换到一种或另一种状态以及这如何发挥作用缺乏定植和感染。该提案将研究 RNA 修饰 N6- 甲基腺苷 (m6A) 可以介导白色念珠菌的白色不透明转换和交配。初步研究我和我的合作者发现，缺乏 IME4（m6A 甲基转移酶）的细胞在温度引起不透明到白色的转换，并具有更高的交配效率。作为一个拥有具有念珠菌白色不透明转换和 m6A 生物学的背景，我有独特的资格来研究 m6A 调节介导诱导表型变化的环境信号的假设介导白色念珠菌感染。目标 1 是确定白色不透明物质中 m6A 修饰的目标通过使用 eCLIP 映射 m6A 修饰的单核苷酸位点和 MRB1（m6A）的结合位点进行切换阅读器，以及野生型和 ime4 突变体白色和不透明细胞的基因表达分析。目标2将通过生成基因表达来确定环境如何调节 m6A 的转换和丝化以及细胞在高温或二氧化碳等环境条件下的代谢特征白色不透明的开关或丝化。目标 3 将研究 m6A 在白色念珠菌口腔定植中的作用通过使用 OPC 的细胞培养物和小鼠模型。白色-不透明转换调节口腔定植，我们假设 IME4 在此过程中发挥作用。为了这个目标，我还将研究辐射如何影响念珠菌缺乏 IME4 的细胞及其如何影响小鼠口腔生态位的定植。在目标 4 中，我将研究该函数 m6A 与口腔微生物组的相互作用。我发现的许多 m6A 修饰基因都涉及在种间相互作用中，表明 IME4 和 m6A 调节宿主内的这些相互作用。这研究计划将是第一个结合表观转录组学和念珠菌生物学领域来研究m6A如何有助于临床相关真菌物种的生活方式和致病性，有可能为临床相关真菌物种铺平道路 OPC 的新抗真菌药物治疗方法。我为他们提出了一个全面的培训计划该提案的 K99 阶段包括获得生物信息学、代谢组学、动物模型方面的新技能口腔感染、微生物组分析、著名导师咨询委员会的指导和合作者以及关键专业和领导技能的发展。总的来说，这个计划将有利于我在 R00 阶段过渡到独立，并为我作为独立研究员的职业生涯做好准备。