Adaptive Evolution of Candida Biofilms

念珠菌生物膜的适应性进化

• 批准号: 10412590
• 负责人: KATY C KAO
• 金额: $ 13.8万
• 依托单位: SAN JOSE STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-01 至 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10412590
• 关键词: Adhesions; Antifungal Agents; Biological Adaptation; Candida; Candida glabrata; Candidiasis; Cells; Clinical; Color; Development; Disease; Drug resistance; Environment; Essential Genes; Event; Evolution; Exhibits; Fluconazole; Frequencies; Funding; Fungal Drug Resistance; Future; Gene Deletion; Genetic Determinism; Genotype; Goals; Growth; Heterogeneity; High-Throughput Nucleotide Sequencing; Host Defense; Immunocompromised Host; In Vitro; Individual; Kinetics; Knowledge; Laboratories; Lead; Libraries; Maize; Malignant Neoplasms; Maps; Mentorship; Methods; Microbe; Microbial Biofilms; Microbial Drug Resistance; Molecular; Monitor; Mutagenesis; Mutation; Mycoses; North America; Pathogenesis; Pharmaceutical Preparations; Phase; Phenotype; Population; Population Dynamics; Procedures; Process; Property; Public Health; Real-Time Systems; Research; Resistance; Sampling; Seeds; Stress; Structure; Surface; System; Therapeutic; Time; Training; Transposase; Underrepresented Minority; Universities; Work; antimicrobial drug; base; biological systems; candida biofilm; causal variant; combat; commensal microbes; emerging pathogen; environmental stressor; experience; genome sequencing; in vivo; minority trainee; mutant; novel therapeutic intervention; pathogen; pathogenic fungus; pathogenic microbe; resistance gene; resistance mechanism; stressor; therapeutic development; therapeutically effective; transposon sequencing; treatment strategy; undergraduate student; underrepresented minority student

PROJECT SUMMARY There is a critical need to identify important parameters involved in adaptation to environmental stresses in fungal biofilms, in order to develop effective therapeutic strategies against fungal infections. Treatment of fungal infections by Candida spp, including the emerging pathogen Candida glabrata (C. glabrata), remains a clinical challenge especially in immunocompromised individuals. C. glabrata is now the second most frequently isolated Candida spp in North America. In vivo, microbes mostly exist in biofilms, which serve as protective layers. Cells in biofilms exhibit increased resistance to environmental stresses. Since biofilms are an integral part of pathogenesis, the adaptation of fungal pathogens inside a biofilm is an important aspect that requires a deeper understanding. However, existing knowledge of fungal adaptation in biofilms is limited, partly due to the lack of established experimental methods for the long-term propagation of fungal biofilms. With a long-term goal of contributing to the development of therapeutic strategies for candidiasis, the overall objective of the project is to develop a fungal biofilm propagation method for use in experimental adaptive evolution, and to apply the system to identify essential genes involved in C. glabrata biofilms. In Aim 1, a biofilm culture system suitable for long- term in vitro evolution will be developed and characterized. A key feature of the method being developed is the ability to grow multiple biofilms from the same seed biofilm, allowing multiple procedures for analysis and characterization of biofilms at each passage during in vitro evolution. Aim 2 will Implement the fungal biofilm propagation system for in vitro evolution of C. glabrata to environmental stressors. Key properties, such as amount of biofilm formation and changes in biofilm structure will be monitored. Molecular mechanisms associated with adaptation to environmental stressors in fungal biofilms will be elucidated based on genome- sequencing and phenotypic analyses. Aim 3 will combine the biofilm propagation method with transposon sequencing to identify essential genes involved in biofilm formation in C. glabrata. The method being developed can be broadly applied to other microbial pathogens to better identify how pathogens adapt and evolve in a more host-relevant environment, and enables the identification of potential therapeutic strategies against difficult-to- treat biofilms. This R16 will fund the research of approximately 20 undergraduates (UGs) over four years, including many minority trainees. UGs will perform the majority of the proposed work, with training and mentorship from a technician and the PI. This funding would allow the PI to continue to develop a strong track record in research, give meaningful research experiences to underrepresented minority students, and enhance the research capacity at San Jose State University.

项目摘要 目前迫切需要确定真菌适应环境胁迫的重要参数， 生物膜，以便开发针对真菌感染的有效治疗策略。治疗真菌 念珠菌属感染，包括新兴病原体光滑念珠菌（C. glabrata），仍然是一个临床 尤其是免疫功能低下的人。C. glabrata现在是第二个最经常被隔离的 北美的念珠菌属。在体内，微生物大多存在于生物膜中，生物膜起保护层的作用。细胞 在生物膜中表现出对环境应力的增加的抗性。由于生物膜是生物膜的组成部分， 致病机制，适应真菌病原体内的生物膜是一个重要的方面，需要更深入的研究 认识然而，生物膜中真菌适应的现有知识是有限的，部分原因是缺乏 建立了真菌生物膜长期繁殖的实验方法。长期目标是 为促进念珠菌病治疗策略的发展，该项目的总体目标是 开发用于实验适应性进化的真菌生物膜繁殖方法，并应用该系统 以鉴定C.光滑生物膜。在目的1中，提供了适合于长时间培养的生物膜培养系统。 术语体外进化将被开发和表征。正在开发的方法的一个关键特征是， 从同一种子生物膜生长多个生物膜的能力，允许多个分析程序， 在体外进化过程中每次传代时生物膜的表征。目的2实现真菌生物膜 C.离体进化的繁殖体系光滑植物对环境压力源的反应。关键属性，例如 将监测生物膜形成的量和生物膜结构的变化。分子机制 与真菌生物膜中对环境压力源的适应相关的基因组- 测序和表型分析。目的3将生物膜增殖法与转座子联合收割机相结合 测序以鉴定参与C.光滑的正在开发的方法 可以广泛应用于其他微生物病原体，以更好地识别病原体如何适应和进化， 宿主相关的环境，并能够识别潜在的治疗策略， 处理生物膜。这个R16将在四年内资助大约20名本科生的研究， 包括许多少数民族学员。UGS将执行大部分拟议的工作，并提供培训， 技术人员和PI的指导。这笔资金将使PI能够继续发展强大的轨道 研究记录，为代表性不足的少数民族学生提供有意义的研究经验，并提高 圣何塞州立大学的研究能力。