Recombinant Expression of Human Type 1 Collagen for Tissue Engineering and Biomaterials

用于组织工程和生物材料的人 1 型胶原蛋白的重组表达

• 批准号: 10325908
• 负责人: Genevieve M. Vidanes
• 金额: $ 28.79万
• 依托单位: PROVENANCE GROUP, INC.
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-01 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10325908
• 关键词: 3-Dimensional; ART protein; Adhesions; Adoption; Animals; Biocompatible Materials; Biological Assay; Biomanufacturing; Bioreactors; Cattle; Cell-Matrix Junction; Cells; Collagen; Collagen Fibril; Collagen Type I; Controlled Environment; Engineering; Enzymes; Escherichia coli; Extracellular Matrix; Face; Family suidae; Fermentation; Foundations; Goals; Human; Hydroxylation; Lead; Length; Mass Spectrum Analysis; Methodology; Mixed Function Oxygenases; Modeling; Modification; Molecular; Monitor; Organ; Organism; Organoids; Phase; Plants; Post-Translational Protein Processing; Price; Process; Procollagen-Proline Dioxygenase; Production; Productivity; Proline; Property; Proteins; Proteolysis; Recombinant Proteins; Recombinants; Research Personnel; Risk; Role; Safety; Skin; Small Business Innovation Research Grant; Source; Specificity; Spectrum Analysis; Structure; System; Technology; Testing; Therapeutic; Time; Tissue Engineering; Toxic effect; Transgenic Organisms; Transgenic Plants; Western Blotting; Work; Yeasts; base; biomaterial compatibility; bioprinting; bone; cost; cost effective; density; flasks; human tissue; immunogenicity; innovation; migration; novel; pathogen; peroxisome; prevent; protein folding; public health relevance; repaired; research and development; scaffold; scale up; tissue reconstruction

Abstract Collagen forms a highly organized, three-dimensional network that facilitates cell attachment, migration, proliferation and differentiation. Strength, persistence and biocompatibility are the inherent properties of collagens that make them well suited as scaffolds in tissue engineering efforts for repair and reconstruction of tissue, bone, and skin. Currently, the primary source of these collagens is animal derived, which causes concerns (reliability, safety, inconsistency, etc). While recombinant production of human collagens promises the most likely solution, the state of art protein production technologies suffer inefficiencies in the production of full length, functional collagens. As such, there’s a recognizable gap between the demand for recombinant collagens and the supply thereof. Provenance has devised a unique strategy for overcoming the limitations of these recombinant protein production systems. We propose to use our novel platform to efficiently synthesize full length human type I collagens with the required post translational modifications at high titers. We will demonstrate the recombinant human type I collagen resembles its native counterpart. We propose to generate highly productive strains expressing full length human type I collagen, that shows the appropriate prolyl hydroxylation, and optimize their production processes at different scales (shake flasks, bioreactors). Ultimately, the approach described here should lead to a cost effective production strategy for recombinant human type I collagen, and alleviate the severe limitations in the availability of this collagen for tissue engineering, and as biomaterials in other biomedical applications. In addition, the availability of an inexpensive source of human type I collagen should allow researchers to expand the role of collagen as a biomaterial for tissue engineering applications. Successful completion of the work proposed in this Phase I proposal can open avenues towards tailoring properties of collagens, a long term goal to impart new functionalities which in turn can lead to novel applications in therapeutic and biomedical applications.

摘要 胶原蛋白形成一个高度组织的三维网络，促进细胞附着、迁移， 增殖分化。强度、持久性和生物相容性是 胶原蛋白，使其成为组织工程修复和重建的支架 组织、骨骼和皮肤。目前，这些胶原蛋白的主要来源是动物来源的，这导致 关注事项(可靠性、安全性、不一致性等)。而重组人胶原蛋白的生产承诺 最有可能的解决方案是，最先进的蛋白质生产技术在生产 全长的功能性胶原蛋白。因此，在重组人的需求之间存在着明显的差距 胶原蛋白及其供应。Provenance设计了一种独特的策略来克服 这些重组蛋白生产系统。我们建议使用我们的新平台来高效地合成 全长人类I型胶原蛋白与所需的翻译后修饰在高滴度。我们会 证明重组人I型胶原蛋白与天然胶原蛋白相似。我们建议生成 高效表达全长人I型胶原的菌株，显示出适当的脯氨酸 羟基化，并在不同规模上优化其生产工艺(摇瓶、生物反应器)。 最终，这里描述的方法应该会导致具有成本效益的重组人生产策略 人I型胶原蛋白，并缓解这种胶原蛋白在组织中可获得性的严重限制 以及在其他生物医学应用中作为生物材料。此外，还提供了一款价格低廉的 人类I型胶原的来源应该允许研究人员扩大胶原作为生物材料的作用 组织工程应用。成功完成此第一阶段提案中提议的工作可以 为调整胶原蛋白的特性开辟了新的途径，这是一个长期目标，旨在赋予新的功能 TURN可以在治疗和生物医学应用方面带来新的应用。