Antifungal Imunoliposomes

抗真菌免疫脂质体

基本信息

批准号：
9926213
负责人：
Richard Brian Meagher
金额：
$ 18.88万
依托单位：
UNIVERSITY OF GEORGIA
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-05-06 至 2022-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9926213
关键词：
3-Dimensional Aftercare Amphotericin B Antibodies Antifungal Agents Antifungal Therapy Antigens Apical Area Aspergillosis Aspergillus Aspergillus fumigatus Asthma Azoles Binding Biochemical Biochemistry Biological Biological Assay Biology Caspofungin Cell Count Cell Survival Cell Wall Cells Cessation of life Chronic Obstructive Airway Disease Cryptococcosis Cystic Fibrosis Cytotoxic agent Development Dose Drug Delivery Systems Drug Targeting Enzyme-Linked Immunosorbent Assay Fluorescence Microscopy Formulation Germ Goals Hyphae Immune system Immunize Immunocompromised Host Immunoliposome Immunologic Receptors Immunology In Vitro Industrial fungicide Infection Life Light Liposomes Lung Lung diseases Mammalian Cell Membrane Methods Microscopy Monoclonal Antibodies Mus Mycoses Organ Outcome Pathogenicity Pathology Patient-Focused Outcomes Patients Pattern Pharmaceutical Preparations Pharmacotherapy Protein Chemistry Proteins Protocols documentation Pulmonary Pathology RNA Reagent Refractory Risk Science Scientist Specificity Structure of parenchyma of lung Surface Antigens Survival Rate Testing Therapeutic Toxic effect Treatment Efficacy Tube Uncertainty beta-Glucans cancer cell cancer therapy cell type comparative efficacy cytotoxicity dectin 1 effectiveness evaluation efficacy testing exosome experimental study fungus improved improved outcome in vivo innovation meetings mouse model pathogenic fungus prevent public health relevance screening side effect targeted delivery targeted treatment uptake

项目摘要

Project Summary/Abstract Invasive fungal infections cause millions of deaths each year and they are refractory to treatment. Patients at the greatest risk of developing life-threatening fungal infections have weakened immune systems or various other lung disorders. Aspergillus fumigatus and related fungal species cause pulmonary invasive aspergillosis. Even after treatment with antifungals such as amphotericin B (AmB) packaged in liposomes, one-year survival among immunocompromised patients with invasive aspergillosis is only 25 to 60%. Antifungals are toxic to patients with numerous side effects and no current treatment clears the lungs of fungi. Without a doubt, there is room for improving antifungal therapeutics to treat aspergillosis. Immunoliposomes are liposomes coated with a monoclonal antibody (mAb) or other protein with specificity for a surface antigen on target cells, enabling the active delivery of a therapeutic drug to a specific cell type. Drug loaded immunoliposomes greatly improve cell-type specificity and reduce cytotoxicity such that higher doses of drug may be delivered to target cells than with liposomes. The overarching goal of this exploratory proposal is to demonstrate that fungal-cell wall targeted immunoliposomes loaded with antifungal agents have increased efficacy over drug loaded liposomes. Our experimental hypothesis is that AmB loaded immunoliposomes coated with monoclonal antibodies or Dectin-1 targeting A. fumigatus cell wall antigens will show enhanced fungal-specific delivery and antifungal activity over untargeted liposomes. Two innovations further our goal. First, we isolated a battery of A. fumigatus cell-wall specific mAbs that bind differentially to various regions on hyphae, germ tubes and conidia, that will enable targeted delivery of immunoliposomes to fungi in vitro and in vivo. Second, we furthered the development of simplified protocols for constructing small batches of fluorescent AmB immunoliposomes and screening methods that will accelerate our examining various targeted-liposomal formulations to fungi in vitro and in vivo in the lungs of infected mice. Our first specific aim is to screen among mAbs and Dectin-1 for targeting protein constructs that achieve the most efficient uptake of fluorescent AmB immunoliposomes into A. fumigatus cells. Results will be quantified by fluorescent microscopy, ELISA, and by assaying fungal cell survival. Our second specific aim is to show that treating infected mice with AmB loaded immunoliposomes targeting A. fumigatus cell wall component(s) kills more fungal cells and increases mouse survival rates relative to untargeted drug loaded liposomes. Results will be evaluated by examining the (1) reduction in A. fumigatus viable cell numbers, (2) alteration in lung pathology, and (3) improvement in mouse survival rates. A team of well acquainted scientists with expertise in immunology, immunoliposomes, fungal biology and pathology, and biochemistry has been assembled.

项目摘要/摘要侵入性真菌感染每年会导致数百万人死亡，并且对治疗难治性。患者出于最大的风险发展威胁生命的真菌感染已削弱免疫系统或各种其他肺部疾病。曲霉菌和相关的真菌物种会引起肺侵袭性曲霉病。即使在用脂质体中包装的两性霉素B（AMB）等抗真菌治疗后，一年生存在侵袭性曲霉病的免疫功能低下的患者中，仅为25至60％。抗真菌对副作用且没有治疗的患者清除了真菌的肺。毫无疑问，有改善抗真菌疗法治疗曲霉病的空间。免疫脂质体是涂有单克隆抗体（MAB）或其他具有特异性的蛋白质的脂质体对于靶细胞上的表面抗原，可以使治疗药物的主动输送到特定的细胞类型。药物加载的免疫脂质体大大提高了细胞型特异性并降低细胞毒性，使得更高与脂质体相比，可以将药物剂量输送到靶细胞。此探索的总体目标建议是证明带有抗真菌剂的真菌壁靶向免疫膜体的对药物载荷脂质体的功效提高。我们的实验假设是AMB已加载涂有单克隆抗体或dectin-1靶向烟曲霉细胞壁抗原的免疫脂质体将在未靶向的脂质体上显示出增强的真菌特异性递送和抗真菌活性。我们的目标进一步进一步创新。首先，我们隔离了一组烟曲霉细胞壁特定的mAb 与菌丝，细菌管和分生孢子上的各个区域有区别的结合，这将有针对性地递送在体外和体内对真菌的免疫脂质体。其次，我们进一步开发了简化协议构建一小部分荧光AMB免疫体和筛查方法，该方法将加速我们检查了在感染小鼠的肺中对真菌和体内进行真菌的各种靶向脂质体制剂。我们的第一个具体目的是筛选mAb和dectin-1，以靶向实现蛋白质构建体荧光AMB免疫膜体对烟曲霉细胞的最有效摄取。结果将是通过荧光显微镜，ELISA和测定真菌细胞存活来量化。我们的第二个具体目标是为了证明，用靶向烟曲霉的AMBANE的免疫脂质体治疗感染的小鼠组件杀死更多的真菌细胞，并增加小鼠的存活率相对于未靶向的药物负载脂质体。将通过检查烟曲霉的（1）降低可行细胞数量来评估结果，（2）肺病理的改变，（3）小鼠存活率的提高。一个熟悉的科学家团队具有免疫学，免疫脂质体，真菌生物学和病理学以及生物化学方面的专业知识组装。