Critical Factors Influencing Echinocandin Resistance in Candida glabrata

影响光滑念珠菌棘白菌素耐药性的关键因素

• 批准号: 10736488
• 负责人: David S Perlin
• 金额: $ 85.7万
• 依托单位: HACKENSACK UNIVERSITY MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-08-01 至 2028-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10736488
• 关键词: Anabolism; Antifungal Agents; Azoles; COVID-19 patient; Candida; Candida albicans; Candida auris; Candida glabrata; Candidate Disease Gene; Candidiasis; Caspofungin; Cells; Cessation of life; Clinical; Comet Assay; Complex; DNA; Dose; Drug Exposure; Drug Metabolic Detoxication; Drug Targeting; Drug Tolerance; Drug resistance; Electron Spin Resonance Spectroscopy; Electrons; Environment; Epidemiology; Evolution; Exposure to; Gastrointestinal tract structure; Genes; Genetic; Genetic Determinism; Grant; HIV; Health; Heritability; Immunity; In Vitro; Incidence; Individual; Infection; Intervention; Knowledge; Libraries; Life; Light; Macrophage; Malaria; Mass Spectrum Analysis; Measures; Modeling; Mus; Mutation; Mycoses; Oxidants; Pathway interactions; Patients; Pharmaceutical Preparations; Physiological; Population; Predisposition; Premature Infant; Proteins; Reactive Oxygen Species; Reporter; Resistance; Role; Route; Source; Sphingolipids; Survivors; Transplant Recipients; clinically relevant; design; drug action; echinocandin resistance; experimental study; fitness; fungus; gastrointestinal; glucan synthase; in vivo; inhibitor; insight; multidisciplinary; mutant; next generation; pathogen; pathogenic fungus; prevent; resistance mutation; response; screening

Fungal pathogens cause a tremendous health burden worldwide and are associated with over 1.5 million deaths per year globally, eclipsing or equaling malaria, TB, or HIV. The last several decades have seen a shift in the epidemiology of Candida infections, with a decreased incidence of pan-sensitive C. albicans and a concomitant increase in non-albicans Candida species, such as C. glabrata and C. parapsilosis, which either have reduced intrinsic sensitivity to commonly used antifungals or can rapidly acquire drug-resistant mutations. Echinocandins are inhibitors of b-glucan synthase (GS) that are cidal in Candida and well tolerated by patients and are thus used as first-line agents for invasive Candida infections, especially in light of increasing insensitivity to azoles in species such as C. glabrata. However, resistance to echinocandins is also increasing, and in C. glabrata it is the highest among major Candida pathogens (including C. auris). Clinical echinocandin resistance (ECHR) is associated with mutations in genes encoding GS, FKS1 and FKS2, and C. glabrata can rapidly evolve ECHR mutations both in vitro and during patient treatment. However, the cellular mechanisms underpinning this rapid emergence of resistance are not well understood. Previously, we described a universal paradigm for fungi, in which drug exposure by a cidal agent effectively kills susceptible cells but leaves a sub-population of viable non- growing cells that are drug-tolerant. Ultimately, some drug-tolerant cells escape drug action by forming heritable drug resistant mutations. We have identified the gastrointestinal (GI) tract and macrophages as important in-host niches of echinocandin-tolerant C. glabrata, in which ECHR mutants form. We have also shown that the evolution of resistance to the echinocandin caspofungin in the GI tract is more complex than was previously appreciated, involving formation of mutations in FEN1, a sphingolipid biosynthesis gene. Importantly, we also identified fen1 mutations in multiple clinical isolates, where they contribute to reduced caspofungin susceptibility. Finally, we discovered that reactive oxygen species (ROS) are formed in C. glabrata cells during echinocandin treatment. However, genes involved in ROS detoxification are downregulated, and deletion of several such genes greatly induces the formation of ECHR mutations, suggesting that ROS induction is a programmed C. glabrata response that may promote the emergence of resistance. In this grant we propose to use transposon insertion screening to comprehensively identify C. glabrata vulnerabilities specific to the host niches where echinocandin tolerance and resistance occur (Aim 1), to comprehensively define the clinically relevant mutational routes to tolerance and resistance against multiple echinocandins in the gut (Aim 2), and to use accurate multidisciplinary complementary approaches, including electron paramagnetic/spin resonance (EPR/ESR) and in vivo ROS reporters, to clearly identify the type(s) of ROS, their role(s), and their source(s) in C. glabrata during echinocandin treatment (Aim 3). By identifying and validating cellular vulnerabilities important for drug tolerance, it will be possible to devise intervention strategies downstream that prevent resistance.

真菌病原体在世界范围内造成巨大的健康负担，并与150多万人的死亡有关 每年在全球范围内，超过或等于疟疾、结核病或艾滋病毒。在过去的几十年里，我们看到了 念珠菌感染的流行病学，泛敏感白色念珠菌和伴随的发病率下降 非白念珠菌种类增加，如光滑念珠菌和近平假丝酵母菌，它们都有所减少 对常用抗真菌药物的内在敏感性，或可迅速获得抗药性突变。棘球绦虫 是β-葡聚糖合成酶(GS)的抑制剂，在念珠菌中是杀伤性的，患者很好地耐受，因此 用作侵袭性念珠菌感染的一线药物，特别是在对唑类药物越来越不敏感的情况下 光叶锦鸡儿等种。然而，对棘球菌素的抗药性也在增加，在光滑棘球绦虫中，它是 在主要念珠菌中最高(包括金黄色念珠菌)。临床棘球绦虫耐药性(ECHR)是 与编码GS、FKS1和FKS2基因突变相关的光肩星天牛可以快速进化为ECHR 在体外和患者治疗期间都有突变。然而，支撑这种快速发展的细胞机制 抵抗的出现还没有得到很好的理解。在此之前，我们描述了真菌的通用范例 哪种杀伤剂接触的药物有效地杀死了敏感细胞，但留下了一个存活的非 培养耐药的细胞。最终，一些耐药细胞通过形成可遗传的 抗药性突变。我们已经确定胃肠道和巨噬细胞是重要的宿主。 ECHR突变体形成的耐棘球绦虫的生态位。我们还证明了进化 胃肠道对棘球菌素卡泊芬净的耐药性比之前估计的要复杂得多， 涉及神经鞘脂生物合成基因FEN1的突变形成。重要的是，我们还确定了fen1 多种临床分离株的突变，它们有助于降低卡泊芬净的敏感性。最后，我们 发现在棘球绦虫处理过程中，光肩星毛虫细胞中形成了活性氧物种(ROS)。 然而，参与ROS解毒的基因表达下调，几个这样的基因大量缺失 诱导ECHR突变的形成，表明ROS的诱导是一种程序性的光滑假丝藻反应 这可能会促进阻力的出现。在这项资助中，我们建议使用转座子插入筛选 全面识别耐受棘球绦虫的宿主生态位所特有的光滑线虫脆弱性 和耐药发生(目标1)，以全面定义临床上相关的耐受突变途径 和对肠道中多种棘球菌素的抵抗力(目标2)，并使用精确的多学科 补充方法，包括电子顺磁共振/自旋共振(EPR/ESR)和体内ROS 记者，为了弄清光肩星天牛体内ROS的类型(S)、作用(S)和来源(S)， 棘球绦虫治疗(目标3)。通过识别和验证对药物耐受性很重要的细胞脆弱性， 这将有可能设计出防止阻力的下游干预策略。

项目成果

Fungal Resistance to Echinocandins and the MDR Phenomenon in Candida glabrata.

念珠菌的真菌对棘突的抗性和MDR现象。

• DOI: 10.3390/jof4030105
• 发表时间: 2018-09-01
• 期刊: Journal of fungi (Basel, Switzerland)
• 作者: Healey KR;Perlin DS
• 通讯作者: Perlin DS

Prevalent mutator genotype identified in fungal pathogen Candida glabrata promotes multi-drug resistance.

• DOI: 10.1038/ncomms11128
• 发表时间: 2016-03-29
• 期刊: Nature communications
• 影响因子: 16.6
• 作者: Healey KR;Zhao Y;Perez WB;Lockhart SR;Sobel JD;Farmakiotis D;Kontoyiannis DP;Sanglard D;Taj-Aldeen SJ;Alexander BD;Jimenez-Ortigosa C;Shor E;Perlin DS
• 通讯作者: Perlin DS

Drug-Resistant Fungi: An Emerging Challenge Threatening Our Limited Antifungal Armamentarium.

• DOI: 10.3390/antibiotics9120877
• 发表时间: 2020-12-08
• 期刊: Antibiotics (Basel, Switzerland)
• 作者: Arastehfar A;Gabaldón T;Garcia-Rubio R;Jenks JD;Hoenigl M;Salzer HJF;Ilkit M;Lass-Flörl C;Perlin DS
• 通讯作者: Perlin DS

A Novel, Drug Resistance-Independent, Fluorescence-Based Approach To Measure Mutation Rates in Microbial Pathogens.

一种新颖的、独立于耐药性、基于荧光的方法来测量微生物病原体的突变率。

• DOI: 10.1128/mbio.00120-19
• 发表时间: 2019
• 期刊: mBio
• 影响因子: 6.4
• 作者: Shor,Erika;Schuyler,Jessica;Perlin,DavidS
• 通讯作者: Perlin,DavidS

Beyond tissue concentrations: antifungal penetration at the site of infection.

超越组织浓度：感染部位的抗真菌渗透。

• DOI: 10.1093/mmy/myy067
• 发表时间: 2019
• 期刊: Medical mycology
• 影响因子: 2.9
• 作者: Zhao,Yanan;Prideaux,Brendan;Baistrocchi,Shane;Sheppard,DonaldC;Perlin,DavidS
• 通讯作者: Perlin,DavidS