Antifungal Imunoliposomes

抗真菌免疫脂质体

• 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
• 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

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 侵袭性真菌感染每年导致数百万人死亡，而且难以治疗。病人 最有可能发展为危及生命的真菌感染削弱了免疫系统或各种 其他肺部疾病。烟曲霉菌及相关真菌可引起肺部侵袭性曲霉病。 即使在脂质体中包装的两性霉素B(Amb)等抗真菌药物治疗后，仍可存活一年 在免疫功能低下的侵袭性曲霉病患者中，这一比例仅为25%至60%。抗真菌药物对人体有毒性 有许多副作用且目前没有治疗的患者可以清除肺部的真菌。毫无疑问，有 改善抗真菌疗法治疗曲霉病的空间。 免疫脂质体是包被单抗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，使治疗药物能够主动输送到特定类型的细胞。 载药免疫脂质体大大提高了细胞类型的特异性，降低了细胞毒性，使更高的 剂量的药物可以被输送到靶细胞，而不是脂质体。这次探险的首要目标是 建议证明真菌细胞壁靶向免疫脂质体负载抗真菌药物具有 比载药脂质体更有效。我们的实验假设是Amb加载了 包被针对烟曲霉细胞壁抗原的单抗或Dectin-1的免疫脂质体将 显示增强的真菌特异性递送和抗真菌活性超过非靶向脂质体。 两项创新推动了我们的目标。首先，我们分离了一组烟曲霉细胞壁特异的单抗 与菌丝、芽管和分生孢子上的不同区域有区别地结合，从而实现靶向传递 免疫脂质体对真菌的体外和体内研究。第二，我们进一步开发了简化的协议， 荧光Amb免疫脂质体的小批量构建和筛选方法将加快 我们在体外和体内对感染小鼠肺部的真菌进行了各种靶向脂质体制剂的检测。 我们的第一个特定目标是在单抗和Dectin-1中筛选靶向蛋白结构，以实现 荧光Amb免疫脂质体进入烟曲霉菌细胞的最有效摄取。结果将是 用荧光显微镜、酶联免疫吸附试验和真菌细胞存活率测定。我们的第二个具体目标是 以烟曲霉细胞壁为靶点的Amb免疫脂质体治疗感染小鼠 成分(S)杀死更多真菌细胞并提高小鼠存活率相对于非靶向药物加载 脂质体。结果将通过检查(1)烟曲霉菌活细胞数量的减少来评估，(2) 肺组织病理改变；(3)提高小鼠存活率。一群熟知的科学家 拥有免疫学、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 集合好了。