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.

项目总结/摘要 侵袭性真菌感染每年导致数百万人死亡，并且难以治疗。患者 在发展威胁生命的真菌感染的最大风险削弱了免疫系统或各种 其他肺部疾病。烟曲霉及其相关真菌引起肺侵袭性曲霉病。 即使用脂质体包装的抗真菌药如阿替西霉素B（AmB）治疗后， 在免疫功能低下的侵袭性曲霉病患者中，只有25%到60%。抗真菌药物对 有许多副作用且目前没有治疗方法的患者可以清除肺部的真菌。毫无疑问， 改善抗真菌疗法治疗曲霉病的空间。 免疫脂质体是包被有单克隆抗体（mAb）或其他具有特异性的蛋白质的脂质体 用于靶细胞上的表面抗原，从而能够将治疗药物主动递送至特定细胞类型。 载药免疫脂质体极大地改善了细胞类型特异性并降低了细胞毒性，使得较高的细胞毒活性。 与脂质体相比，可以将一定剂量的药物递送至靶细胞。本探索性研究的总体目标是 一项提议是证明，载有抗真菌剂的真菌细胞壁靶向免疫脂质体具有 相对于载药脂质体的增加的功效。我们的实验假设是AmB负载 免疫脂质体包被单克隆抗体或Dectin-1靶向A.烟曲霉细胞壁抗原将 显示出比非靶向脂质体增强的真菌特异性递送和抗真菌活性。 两项创新推动了我们的目标。首先，我们分离出一组A。烟曲霉细胞壁特异性mAb， 与菌丝、芽管和分生孢子上的不同区域有差异地结合，这将使得能够靶向递送 免疫脂质体在体外和体内对真菌的作用。其次，我们进一步开发了简化的协议， 构建小批量的荧光AmB免疫脂质体和筛选方法， 我们研究了各种靶向脂质体制剂在体外和体内感染小鼠肺部的真菌。 我们的第一个具体目标是在mAb和Dectin-1中筛选靶向蛋白构建体， 荧光AmB免疫脂质体最有效地摄取到A.烟曲霉细胞结果将 通过荧光显微镜、ELISA和通过测定真菌细胞存活来定量。我们的第二个具体目标是 显示用靶向A.烟曲霉细胞壁 相对于非靶向药物负载， 脂质体。将通过检查（1）A的减少来评价结果。烟曲霉活细胞数，（2） 肺病理学的改变，和（3）小鼠存活率的改善。一组熟悉的科学家 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， 组装好了