Understanding and targeting non-genetic mechanisms of drug resistance

了解和针对耐药性的非遗传机制

• 批准号: 10590490
• 负责人: Jing Lin Xie
• 金额: $ 12.11万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-06-14 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10590490
• 关键词: Aftercare; Antifungal Agents; Biological; Biology; Brefeldin A; CRISPR interference; CRISPR/Cas technology; Candida albicans; Cells; Chemicals; Chromatin; Clinical; Clinical Microbiology; Collaborations; Collection; Communicable Diseases; Communities; DNA Sequence; Development; Development Plans; Drug Tolerance; Drug resistance; Educational process of instructing; Environment; Epigenetic Process; Evolution; Exposure to; Fluconazole; Fluconazole resistance; Fostering; Foundations; Frequencies; Gene Expression; Gene Expression Profile; Genetic; Genetic Transcription; Goals; Growth; Health Care Costs; Heritability; Heterogeneity; Human; Human Pathology; Infrastructure; Kinetics; Laboratories; Libraries; Maintenance; Malignant Neoplasms; Measurement; Memory; Mentors; Methodology; Modeling; Molecular; Mutation; Outcome; Patient-Focused Outcomes; Pharmaceutical Preparations; Phase; Phenocopy; Phenotype; Play; Population; Prevalence; Proteins; Public Health; Research; Research Personnel; Research Training; Resistance; Series; Speed; Stress; Systems Biology; Technology; Testing; Therapeutic; Training; Treatment Failure; Universities; Variant; Work; Writing; career development; clinically relevant; combat; design; droplet sequencing; epigenetic memory; epigenetic variation; fitness; genetic approach; genome-wide; high throughput screening; improved; in silico; infectious disease treatment; insight; medical schools; mutant; non-genetic; novel; novel therapeutic intervention; novel therapeutics; overexpression; pathogenic fungus; pathogenic microbe; programs; proteostasis; research and development; response; single cell technology; single-cell RNA sequencing; skills; small molecule; transcription factor; transcriptome sequencing; yeast prion

PROJECT SUMMARY/ABSTRACT Drug resistance is one of the key contributing factors to treatment failure in infectious disease. Thus, overcoming drug resistance would significantly improve patient outcome and reduce healthcare costs. Extensive research on genetic mechanisms of drug resistance have revealed how drugs are rendered ineffective via changes in the DNA sequence. However, emerging evidence suggest that non-genetic mechanisms such as chromatin- and protein-based epigenetic states may explain how drug resistance arises frequently and rapidly. Understanding the non-genetic mechanisms that drive the evolution of drug resistance could pave the way for the development of novel therapeutic strategies to combat drug resistance. The Research Training Plan will leverage systems biology approaches in a leading human fungal pathogen Candida albicans to dissect the molecular underpinning of pararesistance, a non-genetic mechanism of drug resistance (Aims 1 and 2) and examine the clinical relevance and therapeutic potential of targeting pararesistance (Aim 3). In Aim 1, the applicant Dr. Jing Lin (Lucy) Xie will train in single-cell RNA sequencing and ODE modeling in the K99 phase to identify the transcriptional response and estimate the switching frequency associated with the establishment of pararesistance, developing a quantitative framework to understand how pararesistance is established. In Aim 2, Dr. Xie will train in pooled screen with CRISPRi in the K99 phase and complete the analysis in R00 phase to identify regulators of pararesistance and determine how pararesistance is maintained. In Aim 3, Dr. Xie will establish the prevalence of pararesistance in a collection of ~1,000 wild C. albicans isolates and identify chemical modulators of pararesistance in a library of >1,000 bioactive small molecules to investigate the clinical and therapeutic implications of pararesistance in the R00 phase. The Career Development Plan is designed to provide Dr. Xie with the opportunity and support to acquire additional expertise in single-cell RNA-sequencing and CRISPRi technologies and statistical and modeling methodologies, as well as additional professional development training in teaching, writing, and lab management skills. Mentor Prof. Daniel Jarosz is a leading expert in yeast prions and non-genetic inheritance. Complementary expertise is offered by co-mentor Prof. James Ferrell (quantitative biology), and advisors Prof. Ami Bhatt (clinical microbiology) and Prof. Judith Berman (antifungal drug tolerance and resistance), and collaborators Prof. Michael Hallett (single-cell technologies) and Prof. Rebecca Shapiro (CRISPR technologies). The Stanford University School of Medicine fosters a highly collaborative and supportive research environment and provides excellent infrastructures within a vibrant scientific community. In summary, Dr. Xie will receive high-quality training in research and career development, and is poised to launch her own independent research program. The proposed studies will offer mechanistic insight into non-genetic mechanisms of drug resistance and establish a framework for developing novel therapeutics that would circumvent drug resistance, with important implications for microbial pathogens.

项目摘要/摘要 耐药性是导致传染病治疗失败的关键因素之一。因此，克服 耐药性将大大改善患者的结果并降低医疗保健成本。广泛的研究 关于耐药性的遗传机制，已经揭示了如何通过变化使药物无效 DNA序列。但是，新兴的证据表明，非遗传机制，例如染色质和 基于蛋白质的表观遗传态可以解释耐药性如何频繁，迅速产生。理解 驱动耐药性进化的非遗传机制可能为发展铺平道路 新型治疗策略以打击耐药性。研究培训计划将利用系统 在领先的人类真菌病原体白色念珠菌中，生物学方法剖析了分子的基础 pan抗性，一种耐药性的非遗传机制（目标1和2），并检查临床 靶向促进性的相关性和治疗潜力（AIM 3）。在AIM 1中，申请人Jing Lin博士（露西） XIE将在K99阶段进行单细胞RNA测序和ODE建模，以识别转录 响应并估计与建立panaresistance相关的开关频率，开发 一个定量框架，以了解如何建立panaresistance。在AIM 2中，Xie博士将培训 在K99阶段使用CRISPRI进行屏幕，并在R00阶段完成分析，以识别 吸引力并确定如何维持疗程。在AIM 3中，Xie博士将建立患病率 在约有1,000个野生白色念珠菌分离株的集合中的焦点，并确定的化学调节剂 在> 1,000个生物活性小分子的库中的par症，以研究临床和治疗性 在R00阶段的含义。职业发展计划旨在为Xie博士提供 有机会和支持获得单细胞RNA-severing和Crispri的其他专业知识 技术，统计和建模方法，以及其他专业发展培训 在教学，写作和实验室管理技能中。导师丹尼尔·贾罗斯（Daniel Jarosz）教授是酵母王朝的领先专家 和非遗传继承。互补的专业知识由詹姆斯·费雷尔（James Ferrell）教授提供（定量 生物学）和顾问Ami Bhatt教授（临床微生物学）和Judith Berman教授（抗真菌药物公差 和抵抗），以及合作者迈克尔·哈雷特（Michael Hallett）教授（单细胞技术）和丽贝卡·夏皮罗（Rebecca Shapiro）教授 （CRISPR技术）。斯坦福大学医学院促进了高度合作和支持的 研究环境并在充满活力的科学界提供了出色的基础设施。总之， Xie博士将接受研究和职业发展方面的高质量培训，并准备发起自己的 独立研究计划。拟议的研究将提供对非基因机制的机械洞察力 耐药性并建立了开发新型治疗剂的框架，以规避药物 抗性，对微生物病原体具有重要意义。