The role of sex in the life cycle and transmission of Pneumocystis

性在肺孢子虫生命周期和传播中的作用

• 批准号: 10350565
• 负责人: Melanie T Cushion
• 金额: $ 48.82万
• 依托单位: UNIVERSITY OF CINCINNATI
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-02-01 至 2024-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10350565
• 关键词: 3-Dimensional; Address; Anidulafungin; Antifungal Agents; Biological Assay; Biological Markers; Cell Cycle; Chronic; Clinical; Competence; Complex; Data; Diagnosis; Dose; Electron Microscopy; Electrons; Evaluation; Exposure to; Gene Expression; Genes; HIV; Immunocompromised Host; Immunotherapy; Infection; Interruption; Lead; Life Cycle Stages; Lung; Malignant Neoplasms; Mass Spectrum Analysis; Measurement; Methods; Microbe; Microscopic; Modeling; Morphology; Mus; Organism; Outcome; Partner in relationship; Patients; Pharmaceutical Preparations; Phase; Pneumocystis; Pneumocystis carinii; Pneumonia; Population; Proliferating; Prophylactic treatment; Proteins; Proteomics; Reporting; Rheumatoid Arthritis; Role; Series; Sexual Transmission; Signal Transduction; Techniques; Time; Withdrawal; asexual; base; fungus; genetic signature; mouse model; novel; novel therapeutics; pathogenic fungus; pneumonia treatment; polyglucosan; potential biomarker; prevent; prophylactic; sex; sexual role; targeted agent; therapeutic target; tool; transmission process; treatment duration

Pneumocystis spp. are host obligate fungal pathogens that cause a fatal pneumonia (PCP) in immunocompromised hosts. Few drugs are effective against PCP and there have been no new therapies for its treatment in decades. PCP remains a problem in HIV-infected patients and is emerging as a clinical concern in newly susceptible populations including patients receiving immunotherapy for chronic conditions. We recently reported that treatment of PCP in murine models with the echinocandins, anti-fungal agents that target β-1,3-D-glucan synthesis (BG), depleted the asci which contain BG, but large numbers of forms that do not express BG remained in the infected lungs and were apparently unable to proliferate. Notably, the treated mice could not transmit the infection without asci. The echinocandins prevented PCP when administered prophylactically at higher doses, suggesting that formation of asci via the sexual cycle may be required for a productive infection. Genes associated with sexual replication and cell cycle perturbation comprised the strongest up-regulated signals in P. murina from anidulafungin treated mice. These infections were devoid of asci, suggesting that P. murina attempted to undergo sexual replication, but could not due to a lack of BG. Based on these data, we posit that asci, and thus sexual replication, is required for progression through the Pneumocystis life cycle. The importance of the sexual cycle for survival of Pneumocystis will be investigated using the anidulafungin treatment and prophylactic models. The specific aims address 2 major questions we propose to pursue: Question 1: Is sexual replication required for completion of the life cycle of Pneumocystis? 3 aims will be used to address this question. The first will be to assess the replication competency of P. murina treated with anidulafungin by transferring them to new hosts. Replication will be determined by a combination of microscopic methods, BG measurement, and quantitative PCR. The second aim will use the prophylactic model to allow low level replication to ask whether replication occurs via the sexual cycle, using the same methods. The third aim will determine whether P. murina can survive during prolonged anidulafungin treatment in the same hosts without proceeding through the sexual cycle or if it is eventually cleared. Question 2: At what point in the Pneumocystis life cycle can the infection be transmitted? The sexual cycle returns after withdrawal of anidulafungin. During 10 days to 4 weeks post-cessation, we will track the emergence of the sexual cycle and identify gene signatures, biomarkers and numbers of asci prior to exposing these mice to naïve recipients. Using these data, we will determine at what point in the life cycle transmission takes place and the thresholds necessary for transmission. Selected proteins based on gene expression data will be validated by proteomics as potential biomarkers associated with return of the sexual cycle and transmission. These studies will identify potential vulnerabilities in the Pneumocystis life cycle that could be exploited for interdiction and add to our understanding of its life cycle and transmission.

气囊属。是宿主专有的真菌病原体，导致致命性肺炎（PCP） 免疫功能低下的宿主。很少有药物对PCP有效，并且没有新的疗法 几十年来的治疗。 PCP仍然是HIV感染患者的问题，并且正在成为临床关注的问题 新近易感人群，包括接受慢性病免疫疗法的患者。 我们最近报道说，用棘突（抗真菌剂）在鼠模型中处理pcp，抗真菌剂 靶β-1,3-D-葡聚糖合成（BG）耗尽了包含BG的ASCI，但大量的形式 没有明确的BG保留在感染的肺中，显然无法增殖。值得注意的是，受过治疗 没有ASCI，小鼠无法传播感染。 eChinocandins施用时会阻止PCP 以较高剂量进行预防，表明A可能需要通过性周期形成ASCI 生产感染。与性复制和细胞周期扰动相关的基因完成了 来自Anidulafungin治疗的小鼠的穆利纳（P. Murina）中最强的上调信号。这些感染没有 ASCI，暗示P. Murina试图进行性复制，但不能因为缺乏BG而引起的。 基于这些数据，我们的海报是，ASCI以及性复制是需要进行性复制的 肺刺生命周期。性周期对肺炎生存的重要性将是 使用Anidulafungin治疗和预防模型进行了研究。具体目的地址2专业 我们建议追求的问题：问题1：完成生命周期所需的性复制 肺炎藻？ 3目标将用于解决这个问题。第一个是评估复制 P. Murina的能力通过Anidulafungin治疗，通过将其转移到新宿主中。复制将是 通过微观方法，BG测量和定量PCR的组合确定。第二个 AIM将使用预防模型来允许低级别的复制询问复制是否通过性发生 循环，使用相同的方法。第三个目标将决定穆里纳（P. Murina）是否可以在延长期间生存 在同一宿主中的Anidulafungin治疗而无需进行性周期，或者最终是否最终 清除。问题2：在肺炎胸膜生命周期的什么时候可以传播感染？这 性周期撤回后的回报。在测最后后10天到4周内，我们将跟踪 性周期的出现并鉴定基因特征，生物标志物和ASCI的数量 将这些小鼠暴露于幼稚的接受者。使用这些数据，我们将在生命周期中确定 发生变速箱和传输所需的阈值。基于基因的选定蛋白质 表达数据将通过蛋白质组学验证，因为与性别返回相关的潜在生物标志物 周期和传输。这些研究将确定肺炎藻生命周期中潜在的脆弱性 可以探索拦截，并增加我们对其生命周期和传播的理解。