Aneuploidy and Acquired Antifungal Drug Resistance in Candida species

念珠菌属的非整倍性和获得性抗真菌药物耐药性

• 批准号: 9885981
• 负责人: Anna M. Selmecki
• 金额: $ 46.5万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-12-01 至 2024-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9885981
• 关键词: Aneuploidy; Antifungal Agents; Automobile Driving; Azole resistance; Azoles; Bioinformatics; Candida; Candida albicans; Candida albicans resistance; Candida auris; Cells; Chromosome Arm; Chromosome abnormality; Chromosomes; Clinical; Cryptococcus neoformans; DNA Repair Gene; DNA Sequence; Development; Drug Exposure; Drug resistance; Elements; Engineering; Event; Evolution; Exhibits; Exposure to; Failure; Flow Cytometry; Fluconazole; Fluconazole resistance; Frequencies; Fungal Drug Resistance; Future; Gene Amplification; Generations; Genes; Genetic Recombination; Genome; Genomics; Goals; Hospitals; Human; In Vitro; Infection; Inverted Repeat Sequences; Investigation; Isochromosomes; Karyotype; Lead; Left; Measures; Molecular; Molecular Genetics; Morbidity - disease rate; Multi-Drug Resistance; Mutation; Outcome; Outcome Study; Pharmaceutical Preparations; Pharmacotherapy; Point Mutation; Population; Positioning Attribute; Predisposition; Recurrence; Repetitive Sequence; Reporter; Reproducibility; Research; Research Project Grants; Resistance; Resistant candida; Sepsis; Stress; Structure; System; TAC1 gene; Techniques; Testing; Therapeutic; Time; Work; antimicrobial; arm; base; comparative genomics; digital; experience; experimental study; fitness; genetic approach; genome sequencing; in vivo; interdisciplinary approach; mathematical model; mortality; mouse model; neglect; novel; novel strategies; novel therapeutic intervention; pathogenic fungus; resistance gene; resistance mechanism; resistance mutation; theories; whole genome; yeast genetics

ABSTRACT The rapid emergence of antifungal drug resistance in Candida species poses a severe antimicrobial threat worldwide. Azole antifungals, especially fluconazole, are widely used for treating Candida infections, however, azoles induce genome changes in diverse Candida species and acquired resistance can rapidly emerge. Despite this, we lack a comprehensive understanding of the molecular events that drive the acquisition of azole resistance in real time. Previously, we identified that 50% of fluconazole resistant Candida albicans clinical isolates are aneuploid and some of these aneuploidies cause pan-azole resistance. More recently, aneuploidy- acquired fluconazole resistance has been observed in many diverse human fungal pathogens, yet despite the frequency of these aneuploid events, the mechanism causing them is not known. We have developed cutting- edge multidisciplinary approaches that uniquely position us to define the rate and mechanisms of acquired azole resistance in a rigorous and reproducible way. These approaches include controlled in vitro and in vivo evolution, comprehensive comparative genomics and bioinformatics techniques, and extensive molecular genetic approaches. The aims of this application take a new approach to identifying the mechanisms driving antifungal drug resistance. In aim 1, we will determine the impact of azole stress on the rate and dynamics of acquired azole resistance. This will simultaneously enable us to identify novel and recurrent mechanisms of azole resistance, aneuploidy-derived resistance mechanisms, and the extent to which these mutations cause pan- azole and multi-drug resistance. In aim 2, the frequency of segmental chromosome aneuploidies known to cause resistance across diverse clinical isolates will be determined both in vitro and in vivo. Additionally, the impact of repetitive DNA sequences and DNA repair proteins on the formation of segmental aneuploidies will be determined. Our work is significant because it will identify the frequency, order and trajectory of mutations as they arise in a population and how these mutations cause antifungal drug resistance. Together the outcomes from these studies will identify the mechanisms that underlie how C. albicans acquires resistance and can pave the way for developing therapeutics to reduce the significant morbidity and mortality caused by diverse antifungal drug resistant Candida species.

摘要 念珠菌属真菌耐药性的迅速出现构成了严重的抗菌威胁 国际吧唑类抗真菌药，特别是氟康唑，广泛用于治疗念珠菌感染，然而， 唑类诱导不同念珠菌属物种的基因组变化，获得性耐药性可迅速出现。尽管 因此，我们缺乏对驱动唑类药物获得的分子事件的全面了解 在真实的时间里抵抗。以前，我们发现50%的氟康唑耐药的白色念珠菌临床 分离物是非整倍体，并且这些非整倍体中的一些引起泛唑抗性。最近，非整倍体- 已在许多不同的人类真菌病原体中观察到获得性氟康唑耐药性，然而， 这些非整倍体事件的频率，导致它们的机制尚不清楚。我们开发了切割- 边缘多学科的方法，使我们能够独特地确定获得性唑类的速率和机制 以严格和可重复的方式进行抵抗。这些方法包括受控的体外和体内进化， 全面的比较基因组学和生物信息学技术，以及广泛的分子遗传学， 接近。本申请的目的是采用一种新的方法来确定驱动抗真菌药物的机制。 耐药性在目的1中，我们将确定唑应激对获得性细胞凋亡的速率和动力学的影响。 唑类耐药性这将同时使我们能够确定新的和经常性的机制唑 耐药性、非整倍性来源的耐药机制，以及这些突变引起泛- 唑类和多药耐药。在aim 2中，已知引起染色体非整倍性的节段染色体非整倍性的频率 将在体外和体内测定不同临床分离株的耐药性。此外， DNA重复序列和DNA修复蛋白对节段性非整倍体形成的影响将被 测定我们的工作意义重大，因为它将确定突变的频率，顺序和轨迹， 它们出现在人群中，以及这些突变如何导致抗真菌药物耐药性。共同取得的成果 从这些研究中将确定C.白色念珠菌获得耐药性， 开发治疗方法以降低由各种抗真菌药物引起的显著发病率和死亡率的方法 耐药念珠菌属。