The Impact of SARS-CoV-2 Immune Dysregulation on Antifungal Immunity

SARS-CoV-2 免疫失调对抗真菌免疫的影响

基本信息

批准号：
10668549
负责人：
Sixto Manuel Leal
金额：
$ 66.83万
依托单位：
UNIVERSITY OF ALABAMA AT BIRMINGHAM
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-08-05 至 2023-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10668549
关键词：
2019-nCoV ACE2 Air Allergic Bronchopulmonary Aspergillosis Anabolism Antifungal Agents Aspergillus Aspergillus fumigatus Autopsy Biological Biological Assay Biology Bronchioles Bronchoalveolar Lavage Bronchoalveolar Lavage Fluid Cell Culture Techniques Cell Death Cell Line Chemicals Chemotactic Factors Clustered Regularly Interspaced Short Palindromic Repeats Data Development Enabling Factors Epithelial Cells Evaluation Exhibits Formalin Functional disorder Gene Expression Germination Growth Heme Host Defense Human Hyphae Immune Immune response Immune system Immunity Immunocompetent Immunohistochemistry In Situ Individual Infection Infection prevention Intensive Care Units Interferons Intervention Iron Iron Chelation Knock-out Laboratories Light Liquid substance Lung Mediating Modeling Molds Mus Mycoses Neutrophil Infiltration Nutrient Oxyhemoglobin Paraffin Embedding Pathway interactions Patients Pharmacology Predisposition Production Proteins RIPK1 gene RIPK3 gene Regimen Reproduction spores Respiratory System Risk Role S100A8 gene SARS-CoV-2 infection SARS-CoV-2 variant Sampling Siderophores Specimen Structure of parenchyma of lung Testing Therapeutic Tissues Viral Virus Diseases bronchial epithelium chemokine coronavirus disease cytokine digital extracellular fungus improved in vivo inhibitor innovation knock-down mortality mouse model mutant nano-string neutrophil novel pathogenic fungus pathogenic virus prevent severe COVID-19 transcriptomics

项目摘要

Project summary Unlike most invasive mold infections, COVID Associated Pulmonary Aspergillosis (CAPA) occurs in individuals with otherwise intact immune systems suggesting novel biological mechanisms mediating susceptibility to fungal infection. This proposal seeks to evaluate the novel conceptual advancement that immune responses targeting intracellular viral pathogens compromise essential human neutrophil antifungal effector functions required to kill large extracellular fungal hyphae. In Aim 1, we will utilize BAL samples and autopsy formalin-fixed paraffin embedded (FFPE) lung tissues to characterize the expression of human neutrophil antifungal effectors, epithelial cell necroptosis, iron content, and fungi in the respiratory tract during SARS-CoV-2 and CAPA infection. In Aim 2, we will use primary bronchiolar epithelial cell air-liquid interface (ALI) cultures, CRISPR knockout cell lines (MLKL, RIPK1, RIPK3), chemical inhibitors of cell death pathways, and neutrophil:fungal killing assays to determine the mechanism by which SARS-CoV-2 mediates epithelial cell death, iron release, mold growth, and human neutrophil antifungal effector functions. In Aim 3, we will utilize a novel CAPA infection mouse model, cre-lox knockdown of human ACE2 expression in neutrophils (MRP8-Cre) and lung epithelial cells (CC10-Cre) along with iron-deficient fungal mutant strains (ΔsidA, ΔftrA), to evaluate the in vivo impact of SARS-CoV-2 infection on lung epithelial cell necroptosis, iron release, siderophore-dependent fungal growth, and neutrophil recruitment/activation. We believe that the results of the proposed study will shed new light on the fundamental biology mediating viral-associated secondary mold infection which may enable the development of improved therapeutic regimens that mitigate the risk of developing CAPA.

项目概要与大多数侵袭性霉菌感染不同，新冠肺炎相关肺曲霉病 (CAPA) 发生在具有完整免疫系统的个体表明介导新的生物机制该提案旨在评估新的概念进展：针对细胞内病毒病原体的免疫反应会损害人类必需的中性粒细胞在目标 1 中，我们将利用杀死大型细胞外真菌菌丝所需的抗真菌效应功能。 BAL 样本和尸检福尔马林固定石蜡包埋 (FFPE) 肺组织来表征人中性粒细胞抗真菌效应物、上皮细胞坏死性凋亡、铁含量和真菌的表达在目标 2 中，我们将使用初级细支气管。上皮细胞气液界面 (ALI) 培养物、CRISPR 敲除细胞系（MLKL、RIPK1、RIPK3）、细胞死亡途径的化学抑制剂，以及中性粒细胞：真菌杀灭测定，以确定 SARS-CoV-2 介导上皮细胞死亡、铁释放、霉菌生长和人类在目标 3 中，我们将利用一种新型 CAPA 感染小鼠模型， cre-lox 敲低中性粒细胞 (MRP8-Cre) 和肺上皮细胞中人 ACE2 的表达 (CC10-Cre) 以及缺铁真菌突变株 (ΔsidA、ΔftrA)，以评估 SARS-CoV-2感染对肺上皮细胞坏死性凋亡、铁释放、铁载体依赖性真菌的影响我们相信，拟议研究的结果将会消失。关于介导病毒相关继发霉菌感染的基础生物学的新认识，这可能能够开发改进的治疗方案，降低发生 CAPA 的风险。