Transcriptional networks governing A. fumigatus virulence

控制烟曲霉毒力的转录网络

基本信息

批准号：
10687125
负责人：
Scott G Filler
金额：
$ 65.75万
依托单位：
LUNDQUIST INSTITUTE FOR BIOMEDICAL INNOVATION AT HARBOR-UCLA MEDICAL CENTER
依托单位国家：
美国
项目类别：
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/10687125
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 Fright Gene Targeting Genes Genetic Transcription Genome Goals Growth Hemagglutinin Histoplasma capsulatum Hypoxia Immunocompromised Host In Vitro Incidence Infection Invaded Investigation Life Lung Macrophage 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 genome-wide high risk improved innovation interest lung injury member mortality mouse model mutant novel novel diagnostics novel therapeutics overexpression promoter targeted treatment trait transcription factor transcriptome sequencing

项目摘要

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.

抽象的烟曲霉引起的侵袭性感染正在增加，并且仍然与不可接受的高发病率相关。即使采用新疗法，我们对烟曲霉感染生物学的了解仍然有限。在烟曲霉基因组中预测的基因中，超过 95% 的基因未被表征，只有不到 100 个基因被表征确定控制毒力的基因及其途径至关重要。他们之所以采取行动，是因为他们可以指出治疗和诊断开发的高度优先目标。在烟曲霉 (A. fumigatus) 中鉴定出一个转录调节因子 WrpA，它与白色念珠菌具有有限的同源性我们的初步数据表明 Wor1 和组织胞浆菌 Ryp1 控制着 A. 烟曲霉能够抵抗巨噬细胞的杀伤，在缺氧条件下生长，并侵入和肺部损伤 ΔwrpA 缺失突变体在小鼠侵袭模型中的毒力高度减弱。使用 RNA-seq，我们发现 WrpA 控制烟曲霉中约 15% 的基因表达。基因组，包括多个转录因子基因我们的前提是 WrpA 是一个主调节因子为了支持这一前提，我们对 WrpA 进行了初步研究。调节子已经揭示了三种 WrpA 依赖性转录的新致病性相关功能我们的目标是表征烟曲霉中的 WrpA 调节子并进行鉴定。下游效应基因，其产物通过以下方式介导致病性：1）识别转录因子直接受 WrpA 调节并确定其在致病性中的作用 2) 分析选定的 WrpA- 依赖转录因子并鉴定其下游靶基因；3) 确定功能；由 WrpA 调节子控制的效应基因的研究，并研究它们在毒力中的作用。该提案中描述的实验将使我们能够表征控制的关键转录调节因子 A. fumigatus 致病性，然后使用此信息来识别下游效应基因，即烟曲霉的产物这项工作的结果不仅将提供基础。了解烟曲霉毒力机制，也有望确定新的诊断方法，治疗和疫苗目标。