Develop and Assess mRNA Lipid Nanoparticle Vaccines Against Cryptococcosis

开发并评估针对隐球菌病的 mRNA 脂质纳米颗粒疫苗

• 批准号: 10616313
• 负责人: Xiaorong Lin
• 金额: $ 16.62万
• 依托单位: UNIVERSITY OF GEORGIA
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-01-15 至 2024-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10616313
• 关键词: Acquired Immunodeficiency Syndrome; Actins; Animal Model; Animals; Antibody Formation; Antibody titer measurement; Antifungal Agents; Antifungal Therapy; Antigens; Bacterial Infections; Biochemical; Biology; Brain; COVID-19 vaccine; Cell Culture Techniques; Cell surface; Cells; Central Nervous System Fungal Infections; Central Nervous System Infections; Cessation of life; Chemistry; Clinical; Clinical Trials; Code; Communities; Complex; Cryptococcosis; Cryptococcus; Cryptococcus gattii; Cryptococcus neoformans; Cytoplasm; Data; Diagnosis; Disease; Drug Delivery Systems; Encapsulated; Endoplasmic Reticulum; Ensure; Escherichia coli; Ethics; Exploratory/Developmental Grant; Extracellular Protein; Future; Genetic; Goals; Grant; HIV; Human; Immune response; Immunocompetent; Immunocompromised Host; Immunology; Immunosuppression; In Vitro; Individual; Infection; Injections; Life; Lipids; Liposomes; Mammalian Cell; Measures; Meningoencephalitis; Messenger RNA; Microfluidics; Molecular; Monkeys; Mus; Mycoses; Nature; Nucleic Acids; Organ; Organic solvent product; Outcome; Pathogenesis; Patients; Poly(A) Tail; Productivity; Protein Subunits; Proteins; Public Health; RNA vaccine; Recombinant Proteins; Research; Research Personnel; Role; Safety; Serum; Solvents; Stains; Subunit Vaccines; Technology; Testing; Therapeutic; Therapeutic immunosuppression; Time; Transcript; Translating; Translations; Untranslated Regions; Vaccinated; Vaccination; Vaccine Design; Vaccine Production; Vaccines; Viral; Virus; Whole Cell Vaccine; Work; aqueous; combat; combinatorial; cost; design; efficacy evaluation; experience; extracellular; fighting; fungus; glycation; glycosylation; high risk; immunogenicity; in vivo; lipid nanoparticle; mortality; mouse model; mutant; nanoparticle; nanoparticle delivery; novel; pathogenic fungus; plasmid DNA; preclinical study; prevent; protective effect; red fluorescent protein; vaccine access; vaccine candidate; vaccine delivery; vaccine development; vaccine efficacy; vaccine evaluation; vaccine strategy

Cryptococcus neoformans and C. gattii cause life threatening cryptococcal meningoencephalitis, the most common fungal infection of the central nervous system in the world today. The 12-month mortality rate for individuals with cryptococcal meningoencephalitis is 60% even with the current antifungal treatment. Developing vaccines against cryptococcosis to treat high risk patients remains one of the most urgent and challenging goals to combat this devastating fungal infection. So far, inactivated whole cell vaccines and recombinant protein subunit vaccines are being investigated in preclinical studies. Both inactivated whole cell vaccines or protein subunit vaccines have issues (e.g., safety, technical, cost, and efficacy concerns) and none have made it to clinical trials yet. The emerging and clinically proven vaccine technology based on delivering stabilized mRNAs encoding immunogens packaged in lipid nanoparticles (LNPs) has not been developed for any bacterial or fungal infections. Here we will investigate this technology to develop vaccines against cryptococcal infections. Our research group has experience with liposomes and nucleic acids, and have been working with together on dectin-decorated liposomes for targeted antifungal drug delivery for years. The targeted antifungal delivery with dectin-decorated liposomes has shown great promise in both in vitro and animal models. We have also experience with vaccination against Cryptococcus infection using inactivated cryptococcal mutants. Capitalizing on these experiences, our goal is to produce a highly effective mRNA LNP vaccine for cryptococcosis. We have incorporated several new strategies into our novel mRNA vaccine design. For example, encoded immunogens to be tested are selected based on prior demonstrated efficacy of recombinant proteins to be protective immunogens, high in vivo transcript abundance, extracellular localization, and their roles in fungal pathogenesis. Because each extracellular protein will be produced from an mRNA in the host cell, it will be glycated, and hence, more closely resemble proteins produced by the fungus C. neoformans as compared to recombinant proteins produced by E. coli. To meet our goal, we propose to accomplish the following two specific aims: (i) Construct LNPs carrying the mRNAs encoding distinct candidate C. neoformans protein immunogens. (ii) Test the immunogenicity and durable host protection provided by each mRNA nanoparticle vaccine and a combinatorial vaccine in mouse models of cryptococcosis. Completion of this proposal will likely provide a successful platform for mRNA vaccine against cryptococcosis and in the future other fungal diseases. The exploratory nature of the proposed work and the potential impact it could have on our ability to fight this and other deadly fungal pathogens make this application perfectly fit for the R21 mechanism.

隐孢子虫和C. gattii会导致生命威胁性脑膜脑炎，这是最大的 当今世界中枢神经系统的常见真菌感染。 12个月的死亡率 即使接受当前抗真菌治疗，患有隐球菌脑膜脑炎的个体也为60％。 开发针对隐球菌病以治疗高风险患者的疫苗仍然是最紧迫的疫苗 挑战目标，以打击这种毁灭性的真菌感染。到目前为止，全细胞疫苗灭活， 临床前研究正在研究重组蛋白亚基疫苗。两者都灭活了整个细胞 疫苗或蛋白质亚基疫苗有问题（例如，安全，技术，成本和效率问题），无 已经进行了临床试验。 基于稳定的mRNA编码的新兴和临床证明的疫苗技术 在任何细菌或真菌尚未开发脂质纳米颗粒（LNP）中包装的免疫原子（LNP） 感染。在这里，我们将研究这项技术，以开发针对隐球菌感染的疫苗。我们的 研究小组具有脂质体和核酸的经验，并且一直在合作上 多年来，用于靶向抗真菌药物的dectin装饰脂质体。有针对性的抗真菌递送 在体外和动物模型中，二克蛋白充分的脂质体在体外和动物模型中都表现出了很大的希望。我们也有 使用灭活的隐球菌突变体疫苗接种疫苗接种的经验。 利用这些经验，我们的目标是生产高效的mRNA LNP疫苗 隐球菌病。我们已经将几种新策略纳入了我们的新型mRNA疫苗设计中。为了 例如，根据先前证明的重组效率选择要测试的编码的免疫原子 蛋白质是受保护的免疫原子，体内含量高的蛋白质，细胞外定位及其 在真菌发病机理中的作用。因为每个细胞外蛋白将是由宿主中的mRNA产生的 细胞，它将被糖化，因此，将真菌C. neoformans产生的蛋白质更为类似于 与大肠杆菌产生的重组蛋白相比。为了实现我们的目标，我们建议完成 以下两个具体目的：（i）构建载有编码不同候选C. Neoformans的mRNA的LNP 蛋白质免疫原。 （ii）测试每个MRNA提供的免疫原性和耐用的宿主保护 纳米颗粒疫苗和联合疫苗在小鼠隐球菌病模型中。完成此操作 提案可能会为针对隐球菌的mRNA疫苗提供成功的平台，以后 其他真菌疾病。拟议工作的探索性质及其可能对 我们与这种和其他致命真菌病原体作斗争的能力使该应用非常适合R21 机制。