Transcriptional networks governing A. fumigatus virulence

控制烟曲霉毒力的转录网络

• 批准号: 10365846
• 负责人: Scott G Filler
• 金额: $ 67.11万
• 依托单位: LUNDQUIST INSTITUTE FOR BIOMEDICAL INNOVATION AT HARBOR-UCLA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-22 至 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10365846
• 关键词: Abbreviations; Antifungal Therapy; Aspergillosis; Aspergillus; Aspergillus fumigatus; Attenuated; Azole resistance; Binding; Biology; Bone Marrow; Candida albicans; Cell Communication; Cells; ChIP-seq; Data; Defect; Development; Diagnosis; Diagnostic; Disease; EMSA; Electrophoretic Mobility Shift Assay; Epidermal Growth Factor; Epidermal Growth Factor Receptor; Epithelial Cells; Family; Foundations; Fright; Gene Targeting; Genes; Genetic Transcription; Genome; Goals; Growth; Hemagglutinin; Histoplasma capsulatum; Hypoxia; Immunocompromised Host; In Vitro; Incidence; Infection; Invaded; Investigation; Life; Lung; Mediating; Organism; Pathogenesis; Pathogenicity; Pathway interactions; Population; Predisposition; Prevention; Proteins; Reactive Oxygen Species; Regulon; Response Elements; Role; Testing; Therapeutic; Vaccines; Virulence; Work; chromatin immunoprecipitation; experimental study; fungus; gene product; genome-wide; high risk; improved; innovation; interest; macrophage; member; mortality; mouse model; mutant; novel; novel diagnostics; novel therapeutics; overexpression; promoter; targeted treatment; trait; transcription factor; transcriptome sequencing

Abstract Invasive infections due to Aspergillus fumigatus are increasing and are still associated with unacceptably high mortality, even with new therapies. Our understanding of A. fumigatus infection biology is limited. Among 10,180 predicted genes in the A. fumigatus genome, over 95% are uncharacterized, and fewer than 100 genes have demonstrated roles in virulence. It is critical to identify genes that govern virulence and the pathways in which they act because they can point to high priority targets for therapeutic and diagnostic development. We have identified a transcriptional regulator in A. fumigatus, WrpA, that shares limited homology with Candida albicans Wor1 and Histoplasma capsulatum Ryp1. Our preliminary data indicate that WrpA governs the capacity of A. fumigatus to withstand macrophage killing, grow under hypoxic conditions, and invade and damage pulmonary cells in vitro. ΔwrpA deletion mutants have highly attenuated virulence in the mouse model of invasive aspergillosis. Using RNA-seq, we found that WrpA governs the expression of ~15% of genes in the A. fumigatus genome, including multiple transcription factor genes. Our premise is that the WrpA is a master regulator that governs host cell interactions and virulence. In support of this premise, our initial investigations of the WrpA regulon have already revealed novel pathogenicity-related functions of three WrpA-dependent transcription factors, SrbB, Fcr1, and Ndt80. Our goal is to characterize the WrpA regulon in A. fumigatus and to identify downstream effector genes whose products mediate pathogenicity by: 1) identifying the transcription factors that are directly regulated by WrpA and determining their roles in pathogenicity; 2) analyzing selected WrpA- dependent transcription factors and identifying their downstream target genes; and 3) determining the function of effector genes controlled by the WrpA regulon and investigating their roles in virulence. The results of the experiments described in this proposal will enable us to characterize a key transcriptional regulator that governs A. fumigatus pathogenicity and then use this information to identify downstream effector genes, the products of which mediate host cell interactions and virulence. The results of this work will not only provide foundational understanding of A. fumigatus virulence mechanisms, but also hold promise to identify new diagnostic, therapeutic, and vaccine targets.

抽象的 曲曲霉引起的侵入性感染正在增加，并且仍然与不可接受的高度相关 即使有新疗法，死亡率也是如此。我们对烟曲霉感染生物学的理解是有限的。在10,180中 烟曲霉基因组中的预测基因，超过95％的基因未表征，少于100个基因的基因具有 在病毒中展示了作用。确定控制病毒的基因和其中的途径至关重要 它们之所以如此，是因为他们可以指出热和诊断开发的高优先级目标。我们有 在WRPA的A. fumigatus中确定了一个转录调节剂，该调节剂与白色念珠菌具有有限的同源性 WOR1和组织囊肿RYP1。我们的初步数据表明WRPA控制A的能力。 富马图斯（Fumigatus 细胞体外。 ΔWRPA缺失突变体在侵入性的小鼠模型中具有高度减弱的病毒 曲霉病。使用RNA-Seq，我们发现WRPA控制着烟曲霉中约15％的基因的表达 基因组，包括多个转录因子基因。我们的前提是WRPA是一个主管 控制宿主细胞相互作用和病毒。为了支持这个前提，我们对WRPA的初步调查 Regulon已经揭示了三个WRPA依赖性转录的新型致病性功能 因素，SRBB，FCR1和NDT80。我们的目标是描述A. fumigatus中的WRPA法规并确定 下游效应子基因的产物培养基致病性：1）识别转录因子 由WRPA直接调节并确定其在致病性中的作用； 2）分析选定的WRPA- 依赖转录因子并识别其下游靶基因； 3）确定功能 由WRPA调节控制并研究其在病毒中的作用的效应基因。结果 本提案中描述的实验将使我们能够表征一个控制的关键转录调节器 A. fumigatus致病性，然后使用此信息来识别下游效应基因，即 介导宿主细胞相互作用和病毒。这项工作的结果不仅将提供基础 了解烟曲霉病毒机制，但也有望确定新的诊断， 治疗和疫苗靶标。