Accelerating SARS-CoV2 antiviral drug discovery using next generation 3D bioprinted scaffolds

使用下一代 3D 生物打印支架加速 SARS-CoV2 抗病毒药物的发现

• 批准号: 2452230
• 金额: --
• 依托单位: University of Glasgow
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2020
• 资助国家: 英国
• 起止时间: 2020 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2452230
• 关键词: Accelerating; SARS; CoV2; antiviral; drug

Strategic priority area:Industrial collaborative research. Keywords:Drug discovery, SARS-CoV2, respiratory tissueThe pandemic spread of SARS-CoV2, the principle etiological agent of COVID-19, has identified an urgent need for alternative methodologies for the rapid screening & identification of antiviral compounds that limit the pathogenesis & transmission of zoonotic pathogens;specifically in high-risk category groups susceptible to auto-immune and inflammatory disease.This observation is supported by the inherent delays in vaccine production & global immunization required to protect an immuno-naïve population during a pandemic outbreak.Standard screening approaches for the identification of small molecule inhibitors (SMIs) have relied heavily on two-dimensional (2D) cell-based infection assays.While amenable to high-throughput screening, these 2D systems poorly reflect the three-dimensional physiology,mixed cell-type population,or pharmacokinetic properties of the respiratory airway.Consequently, extensive & time-consuming secondary validation assays are required prior to SMI pre-clinical animal testing.Utilizing primary bronchial cells obtained from healthy donors,which contain a mixture of epithelial, fibroblast & goblet cells,we have shown the differentiation of ciliated 3D respiratory epithelium to readily support SARS-CoV2 infection.While amenable to low-throughput inhibitor studies & secondary assay testing,the use of Transwell technology for the differentiation of respiratory cells is time consuming (minimum 4 weeks),labour intensive, & incompatible with automated imaging techniques required for high-throughput SMI screening.Thus, in the case of emerging pathogens that show tropism for differentiated respiratory epithelia new methodologies are required to facilitate the rapid identification of antiviral compounds directly within respiratory tissue.Recent advances in 3D bioprinting have revolutionized the development & mass production of 'next generation' biomolecular scaffolds that can support the differentiation of multiple cell-types.This project will build on an existing collaboration with Cellbricks, a SME company (https://cellbricks.com) which utilizes a proprietary stereolithography-based bioprinting platform to produce biomolecular scaffolds that support the internal adhesion of cells onto a 3D biopolymer matrix.The use of such scaffolds has many distinct advantages over that of existing Transwell or organoid model systems: (1) Constrained internal dimensions for reproducible cell seeding and downstream infection kinetic assays; (2) Internal 3D micro-bays to promote cell adhesion and differentiation; (3) Optical clarity suitable to high-resolution and high-throughput quantitative imaging; (4) stratification of drugs directly in respiratory tissue established from non-infected high-risk donor groups.The objective of this project is to establish the use of these prefabricated 3D scaffolds in the high-throughput screening,identification & stratification of SMI compounds to SARS-CoV2 directly within respiratory tissue;thereby circumventing the need for time consuming secondary validation assays prior to animal testing in vivo. Trainee and project outcomes:This project will provide extensive training in infectious disease research (MRC-UoG CVR) & biomolecular engineering (Cellbricks).This project will develop key translational research skills applicable to both academia & industry,with clearly defined research aims & expertise pertinent to global health,drug discovery, and precision medicine.Outputs from this project will aid the development of novel methodologies relevant to the identification of antiviral compounds to emerging zoonotic pathogens that will support the career progression of its trainee in academia or industry. Outputs from this project will facilitate the identification and stratification of antiviral inhibitors in the treatment of SARS-CoV2 infected COVID-19 patients from high risk groups.

战略优先领域：工业合作研究。关键词：药物发现，SARS-CoV 2，呼吸道组织SARS-CoV 2的大流行传播，COVID-19的主要病原体，已经确定了对快速筛选和鉴定抗病毒化合物的替代方法的迫切需求，这些化合物限制了人畜共患病病原体的发病机制和传播;特别是在易患自身免疫性和炎症性疾病的高危人群中。这一观察结果得到了疫苗生产固有延迟的支持，用于鉴定小分子抑制剂（SMI）的标准筛选方法严重依赖于基于二维（2D）细胞的感染测定。虽然适合于高通量筛选，但这些2D系统很难反映三维生理学、混合细胞类型群体，因此，在SMI临床前动物试验之前，需要大量且耗时的二次验证测定。利用从健康供体获得的原代支气管细胞，其含有上皮细胞、成纤维细胞和杯状细胞的混合物，我们已经显示纤毛三维呼吸道上皮的分化容易支持SARS-CoV 2感染。虽然适合低通量抑制剂研究和二级分析测试，使用Transwell技术分化呼吸细胞是耗时的（最少4周），劳动密集型，与高通量SMI筛查所需的自动成像技术不兼容。在显示出对分化的呼吸道上皮细胞的向性的新出现的病原体的情况下，需要新的方法来促进直接快速鉴定抗病毒化合物，呼吸组织内。3D生物打印的最新进展彻底改变了“下一代”生物分子支架的开发和大规模生产，可以支持多种细胞类型的分化。该项目将建立在与中小企业公司Cellbricks（https：//www.example.com）的现有合作基础上cellbricks.com，该公司利用专有的立体光刻技术-基于生物打印平台，生产生物分子支架，支持细胞在3D生物聚合物基质上的内部粘附。模型系统：（1）用于可再现的细胞接种和下游感染动力学测定的受限内部尺寸;（2）内部3D微隔间，以促进细胞粘附和分化;（3）光学清晰度，适合于高分辨率和高通量定量成像;（4）药物直接在由未感染的高-该项目的目标是建立这些预制的3D支架在高通量筛选中的用途，识别和分层SMI化合物直接在呼吸组织内SARS-CoV 2;从而避免了在体内动物试验之前进行耗时的二次验证测定的需要。学员和项目成果：该项目将提供传染病研究（MRC-UoG CVR）和生物分子工程（Cellbricks）方面的广泛培训。该项目将培养适用于学术界和工业界的关键转化研究技能，明确定义与全球健康，药物发现和精准医学相关的研究目标和专业知识。该项目的产出将有助于开发与新兴人畜共患病病原体抗病毒化合物相关的新方法，这将支持其学员在学术界或工业界的职业发展。该项目的成果将有助于识别和分层抗病毒抑制剂，用于治疗来自高危人群的SARS-CoV 2感染的COVID-19患者。