Improved Therapeutics and Diagnostics for Pneumocystis Pneumonia

改进肺孢子虫肺炎的治疗和诊断

• 批准号: 10375091
• 负责人: JAY K KOLLS
• 金额: $ 51.07万
• 依托单位: TULANE UNIVERSITY OF LOUISIANA
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-02-01 至 2026-10-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10375091
• 关键词: Acquired Immunodeficiency Syndrome; Africa; Antibodies; Antibody Response; Antigens; Antimicrobial Resistance; Asia; Aspergillosis; Biological Assay; Bronchoscopy; CD4 Lymphocyte Count; CD4 Positive T Lymphocytes; Candida; Child; Clinical; Clustered Regularly Interspaced Short Palindromic Repeats; Complement; Complication; Cyst; Data; Death Rate; Detection; Diagnosis; Diagnostic; Drug Interactions; Far East; Funding; Fungal Proteins; Genes; Genetic Heterogeneity; Glucans; Grant; HIV; HIV Infections; HIV Seronegativity; Hospitalization; Host Defense Mechanism; Housing; Human; Humoral Immunities; Immune Sera; Immunity; Immunization; Immunize; Immunocompromised Host; Immunological Models; Immunotherapy; In Vitro; Infection; Label; Legal patent; Life; Ligase; Liquid substance; Lung; Metabolic; Modeling; Monoclonal Antibodies; Mucous Membrane; Mus; North America; Organism; Patients; Phagocytosis; Pharmaceutical Preparations; Pharyngeal structure; Pneumocystis; Pneumocystis Infections; Pneumocystis carinii Pneumonia; Pneumonia; Prevention; Proteins; Proteome; Pulmonary Inflammation; Resolution; Risk; Sampling; Specific qualifier value; Specimen; Stains; Surface; Swab; Syndrome; Techniques; Technology; Testing; Therapeutic; Treatment Efficacy; Treatment outcome; Trimethoprim-Sulfamethoxazole; Urine; Vaccines; Visualization; Western Asia; Yeasts; cohort; cost; diagnostic assay; experimental study; fungus; immune reconstitution; immunogenicity; improved; mortality; mouse model; novel; polyclonal antibody; prevent; subcutaneous; therapeutic evaluation; transcriptome; transcriptome sequencing; transmission process

Pneumocystis (PC) pneumonia remains a serious complication of HIV infection and other immunocompromised states. Recent ICD9/10 data show that since 2008 there are consistently 14-15,000 hospitalizations per year with an average cost of close to $1B per annum in the US alone. Moreover treatment for PCP has not changed in 25 years and there is concern of anti-microbial resistance and drug:drug interactions with TMP-SMX (the frontline therapy for PCP). The well-known inverse relationship between CD4+ lymphocyte count and the risk of PC infection does not hold all of the answers to mechanisms of host defense against this infection. In the prior funding period, we made significant progress on defining the Pneumocystis transcriptome in both ascus and trophozoite forms as well as the surface proteome of both forms through surface labeling of the fungus using techniques that were developed for Candida. We identified troph proteins such as Meu10, a GPI-anchored protein, as well as ascus specified proteins such as glucan synthetase 1 (GSC1). We were able to clone and express the GSC1 ectodomain in yeast, and immunization of mice with GSC1 resulted in antibodies that stain the surface of the ascus and prevent transmission of Pneumocystis in cohousing experiments. We also made significant progress on diagnostic assays to discriminate colonization from infection. Several groups ae now using quantitative PCR with copy number thresholds, which may be a valid approach. In the prior funding period developed and validated a troph specific assay that selectively eradicates the ascus. Furthermore, RNAseq analysis revealed the troph to be much more metabolically active than the ascus. Thus, we have developed life- form specific assays that will be tested in clinical specimens. We hypothesize that GSC1 immunization can prevent pneumocystis transmission and that mucosal immunization is superior to subcutaneous immunization. We also predict that GSC1 antisera or monoclonal antibodies directed against the GSC1 ectodomain can mitigate Pneumocystis IRIS. Moreover, as our RNAseq analysis has also revealed that the troph is clearly the replicative form of the organism in the lung, we hypothesize that an assay to detect troph specific genes will be diagnostic of PJP due to the detection of the replicative form of the organism. We will also determine the genetic heterogeneity of these target genes in samples from 4 continents. We will test this hypothesis with the following specific aims: Aim 1. Determine the immunogenicity and efficacy of systemic (subcutaneous or IM) versus mucosal GSC1 immunization to prevent PCP. Aim 2. Determine the efficacy of anti-ascus or anti-troph antibodies to treat established PCP and immune reconstitution syndrome (IRIS). Aim 3. Determine and validate life-form specific PCR/CRISPR assays in murine and human samples and assess genetic heterogeneity of these targets in samples from North America, Africa, and western and eastern Asia.

肺孢子虫病(PC)肺炎仍然是HIV感染和其他免疫功能受损的严重并发症 各州。ICD9/10最近的数据显示，自2008年以来，每年持续有14-15,000人住院 仅在美国，每年的平均成本就接近10亿美元。此外，对PCP的治疗并没有改变 在25年内，抗菌素耐药性和药物：药物与TMP-SMX(TMP-SMX)的相互作用受到关注 PCP的一线治疗)。众所周知，CD4+淋巴细胞计数与发病风险呈负相关 PC感染并不是宿主防御这种感染机制的全部答案。在前一次 在资助期间，我们在确定ASCUS和ASCUS中的肺孢子虫转录组方面取得了重大进展 滋养体形态以及两种形态的表面蛋白质组通过表面标记的真菌使用 为假丝酵母菌开发的技术。我们确定了Ttroph蛋白，如GPI锚定的Meu10 蛋白质，以及曲霉特有的蛋白质，如葡聚糖合成酶1(GSC1)。我们能够克隆和 在酵母中表达GSC1胞外区，用GSC1免疫小鼠产生抗体 在共住实验中，它可以保护子囊表面，防止肺孢子虫传播。我们还制作了 在区分定居和感染的诊断试验方面取得了重大进展。现在有几个小组 采用带有拷贝数阈值的定量聚合酶链式反应，可能是一种有效的方法。在上一个资助期 开发并验证了一种有选择地根除Ascus的trph特异性检测方法。此外，RNAseq 分析表明，特罗夫菌的新陈代谢活性比子囊要活跃得多。因此，我们发展了生命-- 形成将在临床标本中进行测试的特定检测方法。我们假设GSC1免疫可以 预防肺孢子虫传播，粘膜免疫优于皮下免疫。 我们还预测，针对GSC1胞外结构域的GSC1抗血清或单抗可以 减轻虹膜肺孢子虫。此外，正如我们的RNAseq分析还显示，特罗夫显然是 在肺中复制的形式，我们假设一种检测troph特异性基因的方法将是 由于检测到有机体的复制形式而诊断PJP。我们还将确定基因 这些靶基因在四大洲样本中的异质性。我们将使用以下内容来验证这一假设 具体目标：目标1.确定全身性(皮下或IM)与 粘膜GSC1免疫预防PCP。目的2.确定抗ASCUS或抗滋养细胞的疗效 用于治疗确诊的PCP和免疫重建综合征(IRIS)的抗体。目标3.确定和验证 小鼠和人类样本中的生命型特异性PCR/CRISPR分析及其遗传异质性评估 来自北美、非洲、西亚和东亚的样本中的目标。