A muco-penetrating biomaterial-based subunit vaccine for programming protective immune responses to SARS-CoV-2

一种基于粘膜穿透生物材料的亚单位疫苗，用于编程针对 SARS-CoV-2 的保护性免疫反应

• 批准号: 10612436
• 负责人: David Scott Wilson
• 金额: $ 24.56万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-01 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10612436
• 关键词: 2019-nCoV; Address; Adjuvant; Animals; Antibodies; Antibody titer measurement; Antigen Targeting; Antigen-Presenting Cells; Antigens; Avidity; B-Lymphocytes; Binding; Biocompatible Materials; Biological; Biological Assay; COVID-19 vaccine; Cells; Clinical; Development; Education; Endosomes; Endothelium; Epithelium; Flow Cytometry; Free Radicals; Goals; Half-Life; Immune; Immune Targeting; Immune response; Immunity; Immunize; In Vitro; Intramuscular; Intramuscular Injections; Intranasal Administration; Irrigation; Label; Ligands; Liquid substance; Location; Lung; Lymph; Lymphoid Tissue; Mannose; Measures; Mediating; Mucosal Immune Responses; Mucosal Immune System; Mucous Membrane; Mucous body substance; Mus; Nasal Epithelium; Nasal cavity; Nose; Pathway interactions; Penetration; Peptides; Polymers; Protein Subunits; Proteins; Residencies; Respiratory Mucosa; Respiratory Tract Infections; SARS-CoV-2 spike protein; Safety; Serum; Signal Transduction; Site; Spleen; Structural Protein; Structure of mucous membrane of nose; Subunit Vaccines; T cell response; T-Cell Activation; T-Lymphocyte; Testing; Thigh structure; Tight Junctions; Time; Tissues; Toll-like receptors; Vaccinated; Vaccination; Vaccine Adjuvant; Vaccines; Viral; Viral Load result; Water; aluminum sulfate; combat; copolymer; design; di-block copolymer; efficacy evaluation; efficacy validation; fluorescence imaging; immunogenicity; lymph nodes; mannose receptor; manufacture; monomer; mouse model; mucosal vaccine; neutralizing antibody; novel; polymerization; pre-clinical; preclinical efficacy; protective efficacy; receptor binding; respiratory; respiratory virus; seasonal influenza; uptake; vaccine access; vaccine candidate; vaccine development; vaccine platform; viral pandemic

1. ABSTRACT/SUMMARY Given that the site of entry of SARS-CoV-2 is the respiratory mucosa, an effective vaccine for SARS-CoV-2 should initiate both humoral and respiratory mucosal immune responses. Although an intranasal subunit vaccine would be an ideal platform for SARS-CoV-2, transport across the nasal mucosa and a lack of safe and effective mucosal vaccine adjuvants thwart the development of a clinically-viable intranasal subunit vaccine. We propose to develop an intranasal vaccine composed of SARS-CoV-2 proteins conjugated to an immunostimulatory biomaterial that overcomes the transport barriers of the nasal mucosa and thus induces protective mucosal and systemic immunity. Our platform is composed of SARS-CoV-2 receptor-binding domain portion (RBD) conjugated to water-soluble polymers, termed MPGAP, that are synthesized from monomers that bind nasal mucus, disrupt endothelial thigh junctions, and target and activate antigen presenting cells (APCs). Thus, when administered intranasally, RBD- MPGAP conjugates should (1) adhere to nasal mucus, increasing residency time at the nasal epithelium, (2) dismantle tight junctions, maximizing paracellular transport to underlying APCs and nasal associated lymphoid tissue, (3) target conjugated RBD to and activate APCs, eliciting APC-derived signals that activate T and B cells. By overcoming the biological barriers of the nasal endothelium and targeting immunostimulatory factors to immune cells, RBD- MPGAP should induce protective mucosal and systemic immunity in the absence of off-target effects. RBD-MPGAP conjugates will be produced, characterized, and their ability to bind nasal mucus, enhance paracellular transport, and target and activate antigen presenting cells will be tested in mice. The neutralizing antibody titer of serum and respiratory fluids from RBD-MPGAP-immunized mice will be assessed via an in-vitro SARS-CoV-2 neutralization assay. Finally, the protective efficacy and durability of the mucosal and systemic immunity elicited by internasal RBD-MPGAP will be investigated in a SARS-CoV-2 mouse model. Completion of this project will validate the preclinical efficacy of an intranasal SARS-CoV-2 subunit vaccine and deliver a platform that could combat numerous other respiratory infections, from seasonal influenza to the next respiratory viral pandemic.

1. 摘要/总结 鉴于 SARS-CoV-2 的进入部位是呼吸道粘膜，有效的 SARS-CoV-2 疫苗应该 启动体液和呼吸道粘膜免疫反应。尽管鼻内亚单位疫苗是一种 SARS-CoV-2 的理想平台、跨鼻粘膜运输以及缺乏安全有效的粘膜疫苗 佐剂阻碍了临床上可行的鼻内亚单位疫苗的开发。我们建议开发一种鼻内 由 SARS-CoV-2 蛋白与免疫刺激生物材料缀合而成的疫苗，克服了 鼻粘膜的运输屏障，从而诱导保护性粘膜和全身免疫。我们的平台是 由与水溶性聚合物缀合的 SARS-CoV-2 受体结合域部分 (RBD) 组成，称为 MPGAP，由结合鼻粘液的单体合成，破坏大腿内皮连接，并靶向和 激活抗原呈递细胞（APC）。因此，当鼻内给药时，RBD-MPGAP 缀合物应该 (1) 粘附鼻粘液，增加在鼻上皮的停留时间，（2）拆除紧密连接，最大化 细胞旁路转运至底层 APC 和鼻相关淋巴组织，(3) 靶向结合的 RBD 并激活 APC，引发 APC 衍生信号，激活 T 细胞和 B 细胞。通过克服鼻腔的生物屏障 RBD-MPGAP 可以诱导内皮细胞和将免疫刺激因子靶向免疫细胞，从而诱导保护性粘膜 和在没有脱靶效应的情况下的系统免疫。 RBD-MPGAP 缀合物将被生产、表征和 它们结合鼻粘液、增强细胞旁运输以及靶向和激活抗原呈递细胞的能力将 在小鼠身上进行了测试。 RBD-MPGAP 免疫小鼠的血清和呼吸道液体中和抗体滴度将 通过体外 SARS-CoV-2 中和测定进行评估。最后，防护效果和耐用性 将在 SARS-CoV-2 小鼠模型中研究鼻内 RBD-MPGAP 引发的粘膜和全身免疫。 该项目的完成将验证鼻内 SARS-CoV-2 亚单位疫苗的临床前功效并提供 一个可以对抗多种其他呼吸道感染的平台，从季节性流感到下一种呼吸道病毒 大流行。