The Translational Response of C. neoformans to Oxidative Stress and Macrophage Phagocytosis.

新型隐球菌对氧化应激和巨噬细胞吞噬作用的翻译反应。

基本信息

批准号：
10317272
负责人：
Charles Joel McManus
金额：
$ 25.11万
依托单位：
STATE UNIVERSITY OF NEW YORK AT BUFFALO
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：
2021-07-01 至 2023-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10317272
关键词：
Acquired Immunodeficiency Syndrome Affect Allografting Antifungal Therapy Bone Marrow Bypass Cause of Death Cellular Stress Cessation of life Collaborations Cryptococcus neoformans Data Dependence Environment Enzymes Eukaryotic Initiation Factor-2 Eukaryotic Initiation Factors Exhibits Future Gene Expression Genes Genetic Transcription Genetic Translation Genomic approach Goals Growth HIV Human Hypersensitivity Individual Infection Initiator Codon Investigation Knowledge Lysosomes Meningitis Messenger RNA Mus Mutagenesis Open Reading Frames Organ Transplantation Outcome Oxidative Stress Oxygen Pathogenesis Patients Phagocytosis Phagolysosome Phosphorylation Phosphotransferases Population Predisposition Process Production Prokaryotic Initiation Factor-2 Reactive Oxygen Species Regulation Repression Research Personnel Resistance Ribosomal RNA Ribosomes Role Saccharomyces cerevisiae Scanning Scheme Stress Testing Therapeutic Time Transcript Transcription Initiation Site Translating Translation Initiation Translational Regulation Translations Transplantation Urease Virulence Factors Work biological adaptation to stress extracellular fungus genome-wide innovation interest macrophage mortality mutant nitrosative stress oxidative damage pathogen pathogenic fungus prevent repaired response ribosome profiling stressor therapeutic development therapeutic target transcription factor transcriptome sequencing translatome

项目摘要

The environmental fungus Cryptococcus neoformans is capable of adapting to the human host, and in susceptible individuals, can cause deadly meningitis that is fatal without antifungal therapy. C. neoformans is responsible for ~15% of AIDS-related deaths worldwide, and is a significant cause of mortality and allograft loss in the transplant population. Interactions with macrophages are important to the outcome of infection. In response to the macrophage intracellular environment, C. neoformans expresses factors that promote survival, including the enzyme urease that prevents acidification of the phago-lysosome, and reactive oxygen stress response factors that detoxify reactive oxygen species and repair oxidative damage. We present compelling preliminary data to suggest that both urease and ROS response effectors are regulated at the level of mRNA translation through the kinase Gcn2. Gcn2 is the sole kinase in C. neoformans that phosphorylates eukaryotic translation initiation factor 2 (eIF2), thereby regulating translation initiation. In this collaboration, the expertise of the Panepinto lab in translational regulation in C. neoformans synergizes with the innovative computational and genomics approaches developed by the McManus lab to investigate the translational response to oxidative stress and perform the first study of translational regulation in C. neoformans during macrophage infection. The proposal consists of two aims. The first aim will use ribosome profiling and RNA-seq to investigate the genome wide translational regulation in response to oxidative stress and determine the consequences of GCN2 deletion on ROS response gene expression and susceptibility to oxidative killing by macrophages. The second Aim will examine the translational regulation employed by C. neoformans during intracellular growth in bone marrow derived macrophages, and its dependence on Gcn2. These studies will provide a comprehensive view of how C. neoformans reprograms its translatome in response to oxidative stress, and during macrophage infection. Due to the fundamental importance of translational control, the processes selected for translation during macrophage adaptation are expected to include potential targets for future therapeutic development.

环境真菌新型隐球菌能够适应人类宿主，在易感个体中，可引起致命性脑膜炎，如果不进行抗真菌治疗，这种脑膜炎就会致命。新型隐球菌导致全世界约 15% 的艾滋病相关死亡，并且是移植群体死亡和同种异体移植损失的重要原因。与巨噬细胞的相互作用对于感染的结果很重要。为了响应巨噬细胞内环境，新型隐球菌表达促进生存的因子，包括防止吞噬溶酶体酸化的脲酶，以及解毒活性氧和修复氧化损伤的活性氧应激反应因子。我们提供令人信服的初步数据表明脲酶和 ROS 反应效应器均通过激酶 Gcn2 在 mRNA 翻译水平上受到调节。 Gcn2 是新型隐球菌中唯一磷酸化真核翻译起始因子 2 (eIF2) 的激酶，从而调节翻译起始。在此次合作中，Panepinto 实验室在新型隐球菌翻译调控方面的专业知识与 McManus 实验室开发的创新计算和基因组学方法相结合，以研究对氧化应激的翻译反应，并开展了首次新型隐球菌翻译调控研究巨噬细胞感染期间。该提案有两个目标。第一个目标将使用核糖体分析和 RNA-seq 研究响应氧化应激的全基因组翻译调控，并确定 GCN2 缺失对 ROS 响应基因表达和巨噬细胞氧化杀伤敏感性的影响。第二个目标将检查新生隐球菌在骨髓源性巨噬细胞的细胞内生长过程中所采用的翻译调节及其对 Gcn2 的依赖性。这些研究将全面了解新型隐球菌如何重新编程其翻译组以应对氧化应激以及巨噬细胞感染期间的情况。由于翻译控制的根本重要性，巨噬细胞适应过程中选择的翻译过程预计将包括未来治疗开发的潜在靶标。