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.

抽象的 胶原蛋白形成高度组织化的三维网络，促进细胞附着、迁移、 增殖和分化是细胞的固有特性。 胶原蛋白使其非常适合作为组织工程修复和重建的支架 目前，这些胶原蛋白的主要来源是动物源性的，这导致了胶原蛋白的产生。 尽管人类胶原蛋白的重组生产有希望，但仍存在一些问题（可靠性、安全性、不一致等）。 最有可能的解决方案是，最先进的蛋白质生产技术在生产中效率低下 因此，重组胶原蛋白的需求之间存在明显的差距。 胶原蛋白及其供应设计了一种独特的策略来克服胶原蛋白的局限性。 我们建议使用我们的新型平台来有效合成这些重组蛋白生产系统。 我们将提供具有所需高滴度翻译后修饰的全长人类 I 型胶原蛋白。 证明重组人 I 型胶原与其天然对应物相似。 表达全长人类 I 型胶原蛋白的高产菌株，显示出适当的脯氨酰 羟基化，并优化不同规模的生产工艺（摇瓶、生物反应器）。 最终，这里描述的方法应该会产生一种具有成本效益的重组生产策略。 人类 I 型胶原蛋白，并缓解组织中这种胶原蛋白可用性的严重限制 工程以及其他生物医学应用中的生物材料。 人类 I 型胶原蛋白的来源应该允许研究人员扩大胶原蛋白作为生物材料的作用 成功完成本第一阶段提案中提出的工作可以。 为定制胶原蛋白的特性开辟了道路，这是赋予新功能的长期目标 转变可以在治疗和生物医学应用中带来新的应用。