BLR&D Research Career Scientist Award Application

BLR

基本信息

批准号：
10451505
负责人：
Melanie T Cushion
金额：
--
依托单位：
CINCINNATI VA MEDICAL CENTER RESEARCH
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-04-01 至 2027-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10451505
关键词：
3-Dimensional Age American Anidulafungin Antifungal Agents Area Arthritis Award Biological Biological Assay Bone Marrow Caring Caspofungin Cell Line Cell Wall Chemicals Chloroquine Chronic Chronic Disease Chronic Obstructive Pulmonary Disease Clinical Collaborations Data Development Dihydrofolate Reductase Inhibitor Disease Dose Drug Screening Elderly Evaluation Female Gene Expression Gene Expression Profiling General Population Genes Goals HIV HIV Seronegativity HIV Seropositivity Healthcare Hematological Disease Human Immune Immune system Immunocompromised Host Immunosuppression Immunotherapy In Vitro Infection Infection prevention International Interruption Knowledge Length Life Cycle Stages Lung Malignant Neoplasms Mammalian Cell Methods Micafungin Microbiology Microscopic Midwestern United States Mission Modeling Morbidity - disease rate Mus Mycoses Ohio Opportunistic Infections Outcome Partner in relationship Patients Pharmaceutical Preparations Phase Pneumocystis Pneumocystis Infections Pneumocystis carinii Pneumocystis carinii Pneumonia Pneumonia Poland Population Portugal Preclinical Drug Development Proliferating Property Prophylactic treatment Publications Rattus Reporting Research Rheumatoid Arthritis Risk Rodent Model Role Scientist Seminal Sexual Reproduction Societies Sphingolipids Stress Sulfamethoxazole Therapeutic Time Toxic effect Trimethoprim-sulfamethoxazole drug resistance United States Up-Regulation Veterans Withdrawal analog antimicrobial asexual base cancer diagnosis candidate identification career celecoxib chronic inflammatory disease clinically relevant comorbidity efficacy testing experimental study fungus high risk immunological status immunosuppressed infected vector rodent knockout gene male metabolic abnormality assessment military veteran mortality mouse model new therapeutic target novel therapeutics pathogen pathogenic fungus polyglucosan pre-clinical prophylactic response screening sex targeted treatment therapy duration tool transcriptome sequencing transmission process tumor necrosis factor-alpha inhibitor

项目摘要

Pneumocystis spp. are 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. Typically, PCP has been associated with patients infected with HIV, however, the fulminate pneumonia, PCP, and colonization with Pneumocystis jirovecii (the species infecting humans) are emerging clinical problems in newly susceptible populations in the general and veterans’ populations including bone marrow recipients; patients receiving immunotherapy for rheumatoid arthritis and other chronic inflammatory diseases; and cancer chemo- and immunotherapies. The life cycle of Pneumocystis is suggested to contain both an asexual replication cycle and a sexual cycle involving mating with subsequent formation of asci containing 8 ascospores (1). During the previous Merit Review, we showed that echinocandin treatment of rodents infected with P. murina and P. carinii, which target β-1,3-D-glucan synthesis (BG), depleted the asci which contain BG but large numbers of non-BG expressing life cycle stages remained in the lungs and were unable to proliferate. We further demonstrated that anidulafungin and caspofungin could prevent infection in a prophylactic model, suggesting that formation of asci via the sexual cycle may be required for a productive infection (2). Analysis of gene expression profiles of P. murina in mice treated with anidulafungin, showed strong upregulation of genes associated with sexual replication, though the resulting infections were devoid of asci, the product of sexual reproduction, suggesting that P. murina attempted to undergo sexual replication, but could not due to a lack of BG. Based on these data, we posited that asci, and thus sexual replication, is required to facilitate progression through the life cycle leading to a productive infection. We further posited that presence of asci is required for transmission of Pneumocystis infection. In the present Merit Review, we will explore 2 critical but unanswered questions that will lead to a deeper knowledge of the life cycle of Pneumocystis, and also suggest potential vulnerabilities for targeted treatment concomitant with anidulafungin therapy: (1) Is sexual replication required for completion of the life cycle of Pneumocystis? Tracking of the replication status of P. murina during prolonged treatment with anidulafungin by global gene analysis, BG content, and microscopic methods will reveal whether the non-BG expressing forms numbers remain: 1) static over time, 2) increase, or 3) decrease; suggesting: 1) the lack of BG blocks replication; 2) that an asexual or alternative replication phase permits survival of the fungi; or 3) the lack of sexual replication results in elimination of the infection. (2) Can sexual replication rebound after cessation of prolonged anidulafungin treatment? Mice will be treated with anidulafungin for up to 8 weeks, with 2 cessation time points. Mice in the cessation groups will be tracked for microscopic, BG content, and gene expression evidence of asci formation and return of the pneumonia while remaining under immunosuppression. Mice in the treated and cessation groups will be evaluated for their ability to transmit the infection and the critical number of asci needed for transmission. All studies will be conducted in male and female mice, recognizing sex as a biological variable. The echinocandins are clinically available in the United States. Current monotherapy with any echinocandin for PCP is not warranted as withdrawal can result in return of the pneumonia. The results of the proposed studies will have immediate clinical relevance by determining the length of time viable Pneumocystis can remain in the lungs with concurrent anidulafungin treatment, providing a rationale for duration of therapy with eradication as the goal. These experiments will also identify whether immunosuppressed mice can transmit the infection after withdrawal of anidulafungin and if there is a critical number of asci needed. Finally, the studies will elaborate the life cycle of Pneumocystis and suggest new target strategies.

气囊属。是在免疫功能下引起致命性肺炎（PCP）的强性真菌病原体主持人。很少有药物对PCP有效，并且几十年来没有新的治疗方法。通常，PCP与感染HIV的患者有关，但是，肺炎，PCP，PCP，和肺炎藻（Jirovecii 在一般和退伍军人人口中新易感人群，包括骨髓受体；接受免疫疗法的类风湿关节炎和其他慢性炎性疾病的患者；和癌症化学和免疫疗法。建议肺炎藻的生命周期包含无性复制周期和一个性周期涉及与随后的ASCI形成8个子肠孢子的交配（1）。期间先前的优点综述，我们表明了对啮齿动物的啮齿动物治疗。靶向β-1,3-d-glucan合成（BG）的Carinii耗尽了包含BG但大量的ASCI 表达生命周期阶段的非BG保留在肺部，无法增殖。我们进一步证明Anidulafungin和Caspofungin可以防止预防模型中的感染，这表明生产性感染可能需要通过性周期形成ASCI（2）。基因分析在用Anidulafungin治疗的小鼠中，穆里纳的表达谱图显示基因的强上调与性复制相关，尽管所产生的感染没有ASCI，但性产物是性的繁殖，表明P. Murina试图进行性复制，但由于缺乏 BG。基于这些数据，我们保证ASCI并因此需要进行性复制才能促进进展通过生命周期导致生产感染。我们进一步贴上了ASCI的存在肺炎胸膜感染的传播。在当前的优点审查中，我们将探索2个批判性但没有得到答复的会导致对肺炎史蒂斯生命周期更深入了解的问题，也提出潜在的与Anidulafungin治疗相关的有针对性治疗的脆弱性：（1）是需要性复制的为了完成肺炎藻的生命周期吗？跟踪P. Murina的复制状态通过全球基因分析，BG含量和微观方法延长治疗，将揭示是否保留非BG表达形式的数字：1）随着时间的推移静态，2）增加或3）减少；建议：1）缺乏BG块复制； 2）无性或替代复制阶段允许真菌的生存；或3）缺乏性复制导致消除感染。（2）可以性停止长期Anidulafungin治疗后的复制反弹？老鼠将接受治疗 Anidulafungin长达8周，并有2个停止时间点。戒烟组中的小鼠将被追踪微观，BG含量和基因表达证据的ASCI形成和肺炎的回归在免疫抑制下保持。将评估经过治疗和戒烟组中的小鼠的能力传递感染和传播所需的ASCI的临界数。所有研究都将在男性和雌性小鼠，将性别识别为生物变量。 echinocandins在临床上可用美国。目前没有任何用于pcp的echinocandin的单一疗法，因为戒断可能导致肺炎恢复。拟议研究的结果将立即具有确定随着并发的Anidulafungin的肺部可行的时间长度可以保留在肺中治疗，为根除治疗的持续时间提供了理由。这些实验也将确定免疫抑制的小鼠是否可以在Anidulafungin撤离后传播感染以及是否在那里是需要的关键数量ASCI。最后，这些研究将详细说明肺炎的生命周期，并建议新的目标策略。