Temporal Adaptation to Antifungal Treatment in Pathogenic Fungi

你会调查什么？真菌是一种适应性极强的微生物，甚至可以适应我们用来杀死它们的抗真菌药物。一些研究已经确定了抗真菌药物有效杀灭真菌的机制，以及抗真菌耐药性的关键机制。众所周知，抗真菌治疗还会导致真菌细胞壁中细胞内ROS的显著变化[Lee&Lee 2018]，并可能对哺乳动物感染的存活率产生矛盾的影响[Lee 2012]。这些研究没有解决的是真菌最初是如何感知抗真菌活性并对其做出反应的。改善我们对这些早期适应抗真菌治疗的理解可以突出真菌对其做出反应的特定细胞压力，从而深入了解与人类健康相关的条件下的基本真菌细胞生物学。因此，这个项目的目的是使用模式酵母酿酒酵母和临床相关的酵母光滑假丝酵母建立真菌细胞对抗真菌药物的反应的时间分布。这个项目的重点是调查随着时间的推移，抗真菌暴露如何改变基因表达和蛋白质翻译。这些数据集将被整合，以确定抗真菌治疗反应的时间模式。你将使用最先进的测序技术来进行基因表达和分子遗传学研究。你将与华莱士实验室的同事一起使用生物信息学和统计软件来分析转录和翻译数据。从时间分布来看，我们将预测真菌是如何感知抗真菌压力的，并使用尖端分子技术对酵母进行基因修饰来测试这些假设。由此产生的酵母菌株将被评估抗真菌敏感性，细胞壁变化，以及宿主-病原体相互作用的变化。您将与Childers实验室的同事一起学习致病酵母菌培养、细胞壁和表型分析以及病原菌相互作用。这个项目将显著提高我们对真菌细胞如何感知和适应抗真菌药物背后的分子机制的理解。你将接受什么培训？你将被培养成一名全面发展的科学家，能够与科学和普通观众进行交流。您将学习可移植的方法：微生物学技术、细胞壁和表型分析，以及现代分子方法，包括CRISPR-Cas9基因编辑。您还将从华莱士实验室学到一项竞争激烈且备受关注的技能：如何处理大数据集、提取RNA以及生物信息学和统计分析方面的最佳实践。接下来会发生什么？完成这个项目后，你将成功地获得生命科学行业极具竞争力的技能。越来越多的人需要经过生物信息学培训的科学家，以及理解“大数据”的能力。您从湿实验室研究活动和数据集处理中获得的知识将帮助您在学术或工业环境中推动有影响力的研究。你在这个项目中学到的沟通、分析和解决问题的技能将可以在不同的就业部门之间转移和竞争。

What will you investigate? Fungi are amazingly adaptable microorganisms and can even adapt to the antifungal agents we use to kill them. Several studies have identified the mechanisms by which antifungals are effective at killing fungi and the key mechanisms for antifungal resistance. Antifungal treatment is also known to induce significant changes in intracellular ROS [Lee & Lee 2018], in fungal cell walls [Hopke 2016] and can have paradoxical effects on survival in mammalian infections [Lee 2012]. What these studies do not address is how fungi initially sense and respond to antifungal activity. Improving our understanding of these early adaptations to antifungal treatment can highlight the specific cellular stresses to which fungi are responding, thus giving insight into fundamental fungal cell biology under conditions relevant to human health. Therefore, the aim of this project is to build a temporal profile of how fungal cells respond to antifungal agents using the model yeast, Saccharomyces cerevisiae, and the clinically-relevant yeast, Candida glabrata. The focus of this project is to investigate how antifungal exposure over time alters gene expression and protein translation. These datasets will be integrated to identify temporal patterns of responses to antifungal treatment. You will use state-ofthe- art sequencing to perform gene expression and molecular genetic investigations. You will work with Wallace lab colleagues to use bioinformatics and statistical software to analyse transcriptomic and translational data. From the temporal profile, we will make predictions of how fungi are sensing antifungal stress and test these hypotheses using cutting-edge molecular techniques to genetically modify yeast. The resulting yeast strains will be assessed for antifungal sensitivity, cell wall alterations, and for variations in host-pathogen interactions. You will work with Childers lab colleagues to learn pathogenic yeast cultivation, cell wall and phenotypic analysis, and hostpathogen interactions. This project should significantly improve our understanding of the molecular mechanisms behind how fungal cells sense and adapt to antifungals. What training will you receive? You will be trained to become a wellrounded scientist who is able to communicate with scientific and general audiences. You will learn transferable methodologies: microbiological techniques, cell wall and phenotypic analysis, andmodern molecular approaches, including CRISPR-Cas9 gene editing. You will also learn a competitive and highly sought skill from the Wallace lab: how to handle large datasets, extract RNA, and best practices in bioinformatics and statistical analysis. What comes next? Upon completing this project, you will have successfully gained highly competitive skills for the life sciences industry. There is a growing demand for scientists with bioinformatics training and the capacity to make sense of 'big data'. The knowledge you gain of wet-lab research activities and dataset handling will help you drive impactful research in academic or industrial settings. The communication, analysis, and problem-solving skills you learn on this project will be transferable and competitive across employment sectors.