Development of nucleic acid-based vaccines against gonorrhea

开发基于核酸的淋病疫苗

• 批准号: 10624940
• 负责人: Lisa Ann Lewis
• 金额: $ 20.94万
• 依托单位: UNIV OF MASSACHUSETTS MED SCH WORCESTER
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-05-19 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10624940
• 关键词: Acceleration; Affect; Animal Model; Antibiotic Resistance; Antibiotics; Antibodies; Antibody Formation; Antibody titer measurement; Antigen Targeting; Antigen-Presenting Cells; Antigens; Antimicrobial Resistance; Attenuated; Biology; Biotechnology; COVID-19 pandemic; Ceftriaxone; Cell Nucleus; Cell division; Chimera organism; Chimeric Proteins; Chlamydia; Clinical; Complement; Complement Membrane Attack Complex; Country; Cryopreservation; Cytoplasm; DNA Vaccines; Data; Development; Disease; Ectopic Pregnancy; Epitopes; Equity; Formulation; Gonorrhea; HIV; Immune response; Immunize; Immunoglobulin A; Immunoglobulin G; Immunology; In Vitro; Incidence; Individual; Infection; Infertility; Inflammation; Lead; Link; Measures; Mediating; Messenger RNA; Microbe; Modeling; Monoclonal Antibodies; Mucous Membrane; Multi-Drug Resistance; Mus; Neisseria gonorrhoeae; Nucleic Acid Vaccines; Nucleic Acids; Peptides; Persons; Play; Proteins; RNA vaccine; Recommendation; Reporting; Resistance; Resistance development; Resources; Role; Serum; Sexually Transmitted Diseases; Sialic Acids; Surface; T cell response; Testing; United States; Vaccine Antigen; Vaccines; Vagina; Virulence; Virulence Factors; Woman; Work; bactericide; cervicovaginal; chronic pelvic pain; combat; cost; density; efficacy evaluation; fast protein liquid chromatography; fighting; immunogenicity; improved; in silico; in vivo; infection burden; lipid nanoparticle; lipooligosaccharide; low income country; mRNA Stability; mRNA delivery; mouse model; mutant; nanoparticle delivery; novel vaccines; optimism; peptidomimetics; phase 1 study; plasmid DNA; pre-clinical; preclinical development; public health priorities; receptor; reproductive tract; response; sialic acid binding Ig-like lectin; sialylation; success; synergism; technology platform; transmission process; trend; vaccine candidate; vaccine delivery; vaccine development; vaccine efficacy; vaccine evaluation; vaccine formulation; vaccine platform; vaginal fluid

ABSTRACT Gonorrhea affects over 87 million people annually globally. In 2019, over 600,000 cases of gonorrhea were reported to the CDC, a 92% increase since the historic low in the US in 2009. However, the true incidence of cases annually in the US is estimated to be ~1.5 million. The spread of antimicrobial resistance has severely limited treatment options – currently, ceftriaxone is the only approved first line of treatment. A safe and effective vaccine against gonorrhea is urgently needed. Lipooligosaccharide (LOS) is the most abundant molecule on the gonococcal surface and plays multifaceted roles in bacterial virulence. A LOS epitope recognized by mAb 2C7 (therefore called the 2C7 epitope) is expressed by >95% of Ng in vivo; Ng mutants that do not express the 2C7 epitope are attenuated in mice. Our group has developed a peptide mimic (mimitope) of the 2C7 epitope, which when configured as a multi-antigen peptide (MAP) elicits bactericidal Abs and attenuates Ng colonization in mice. Further, we have elucidated the mechanism of action of MAP as complement-mediated killing (killing through membrane attack complex), strongly suggesting serum bactericidal activity as a mechanistic correlate of protection. In a collaborative venture with Evaxion Biotech, we screened 30 antigens that were identified by EDEN, Evaxion’s proprietary in silico platform, for immunogenicity, bactericidal activity and efficacy in the mouse vaginal colonization model. These studies identified two cell division proteins as promising candidates based on efficacy in vivo. When configured as a fusion protein (chimera), the cell division proteins showed improved ac- tivity in vivo compared to individual proteins and currently is our lead protein vaccine candidate. DNA and mRNA vaccines have proven very successful in the fight against the Covid-19 pandemic. These platforms elicit high levels of protection, have proven safe, are rapidly adaptable and economical. Here, we will leverage these two newer vaccine platforms to deliver the protective gonococcal antigens. A multivalent vaccine has the advantage of broader strain coverage, improved efficacy and would also raise the barrier for development of resistance. In Aim 1, we will evaluate DNA vaccine constructs that contain the 2C7 LOS mimitope (PEP1) configured as an octameric concatemer (PEP1_OCT), the cell division protein chimera, or a combination of the two. Serum will be assessed for antibody titers and complement-dependent bactericidal activity, as well durability of responses. Comparisons will be made with the protein versions of these vaccines currently under development. Mucosal (vaginal) Ab responses will also be measured. T cell responses will be measured by ELISpot. Based on data in vitro, the lead DNA vaccine formulation(s) will be assessed for efficacy in the mouse vaginal colonization model. In Aim 2 the constructs described in Aim 1 will be expressed using an mRNA vaccine delivery platform. As in Aim 1, we will evaluate Ab titers in serum and vaginal fluid, measure serum bactericidal activity, T cell responses and efficacy of the lead mRNA vaccine formulation(s) in the mouse model of gonorrhea. Successful completion of the work in this proposal will identify a lead gonococcal DNA and/or mRNA vaccine for further pre-clinical development.

抽象的 淋病每年在全球影响超过8700万人。 2019年，超过600,000例淋病是 向疾病预防控制中心报告的，自2009年美国历史低下以来增长了92％。但是， 每年在美国，估计案件约为150万。抗菌耐药性的传播严重 有限的治疗选择 - 目前，头孢曲松是唯一批准的第一道治疗方法。安全有效的 迫切需要针对淋病的疫苗。脂肪糖（LOS）是该分子上最丰富的分子 淋球菌表面并在细菌病毒中扮演多方面的作用。 MAB 2C7识别的LOS表位 （这被称为2C7发作）由体内NG> 95％表示；不表达2C7的NG突变体 表位在小鼠中减弱。我们的小组已经开发了2C7表位的肽模仿（Mimitope） 当配置为多抗原肽（MAP）时，会引起细菌ABS并减弱NG定殖 老鼠。此外，我们已经阐明了MAP的作用机理，作为完全介导的杀戮（杀戮） 通过膜攻击复合物），强烈建议血清细菌活性作为机械 保护。在与Evaxion Biotech的合作企业中，我们筛选了30种由 伊甸园（Eden），在硅平台中的埃德（Eden）专有，用于免疫原性，细菌活性和小鼠的效​​率 阴道定植模型。这些研究将两种细胞分裂蛋白确定为承诺的候选者 体内功效。当配置为融合蛋白（Chimera）时，显示的细胞分裂蛋白改善了AC- 与单个蛋白质相比，体内的潮流是我们的铅蛋白疫苗候选者。 DNA和mRNA 事实证明，疫苗在与19009年大流行的战斗中非常成功。这些平台引起了高度 被证明是安全的保护水平，具有迅速适应性和经济性。在这里，我们将利用这两个 较新的疫苗平台可提供受保护的淋球菌抗原。多价疫苗具有优势 更广泛的应变覆盖范围，提高效率，还将提高阻力发展的障碍。在 AIM 1，我们将评估包含2C7 LOS Mimitope（PEP1）的DNA疫苗构建体 八聚体系（PEP1_OCT），细胞分裂蛋白嵌合体或两者的组合。血清会 评估抗体滴度和补体依赖性细菌活性以及反应的持久性。 将与目前正在开发的这些疫苗的蛋白质版本进行比较。粘膜 （阴道）AB反应也将被测量。 T细胞反应将通过ELISPOT测量。基于数据 体外，将评估铅DNA疫苗公式在小鼠阴道定植模型中的效率。 在AIM 2中，AIM 1中描述的结构将使用mRNA疫苗输送平台表示。如 AIM 1，我们将评估血清和阴道液中的AB滴度，测量血清细菌活性，T细胞反应 并在淋病小鼠模型中易于铅mRNA疫苗公式。成功完成 该提案中的工作将确定铅淋球菌DNA和/或mRNA疫苗，以进行进一步的临床前 发展。