STTR Phase I: Biomanufacture of Novel Heparan Sulfate Glycosaminoglycans

这项小型企业技术转让（STTR）项目的更广泛影响/商业潜力将是将新型糖聚合物（称为糖胺聚糖（GAGs））商业化，用于各种重要的医疗应用。GAGs是许多重要生理和病理生理条件（如肿瘤血管生成、血栓形成）的关键成分；然而，由于缺乏聚糖分析的分析技术和缺乏可用于研究的高质量GAG样品，对这些可能对人类健康产生重大影响的作用进行详细了解是具有挑战性的。GAG在细胞信号级联反应中具有精确的功能作用，这取决于特定的GAG结构组成。特定的GAG组成在不同的组织类型以及不同的发育和生理状态中有所不同。因此，为了获得适当的功能特征并获得可重复的实验结果，GAG样品具有一致和明确的组成是至关重要的。不幸的是，大多数商业上可获得的GAG样本来自动物来源，具有很高的固有变异性，并且没有很好地表征。潜在的污染或不确定因素使得动物源性GAG样品更不适合用于人类治疗。本STTR一期项目拟开发基因工程中国仓鼠卵巢（CHO）细胞，产生具有确定成分的gag。将优化培养条件，以保持GAG成分，同时提供足够商业化的产品产量。一系列的工程细胞系将产生，以制备具有不同成分的gag，模拟自然界中发现的不同成分和不同功能特征。GAG样品将在结构和功能上进行表征，以提供每种类型样品的概况，这将是可靠的和可重复的，因为GAG是在受控条件下生长的细胞制备的。这些样本将成为许多生物和医学领域研究人员的宝贵资源。据认为，质量好价格高。然而，为了帮助降低成本和促进商业化，该提案将测试生物反应器的生长条件，以提高生产能力和效率。使用这些高质量GAG样品的研究将为这些GAG在肿瘤学、脂质代谢、组织再生和血液学等各种重要治疗应用中的使用提供主要证明。

The broader impact/commercial potential of this Small Business Technology Transfer (STTR) project will be to commercialize novel sugar polymers known as glycosaminoglycan (GAGs) for a variety of important medical applications. GAGs are key components in a number of important physiologic and pathophysiologic conditions (e.g., tumor angiogenesis, thrombosis); however, developing a detailed understanding of these roles, which may significantly impact human health, is challenging due to a lack of analytical techniques for glycan analysis and the absence of high quality GAG samples available for research. GAGs have precise functional roles in cell signaling cascades, which can vary depending on the particular GAG structural composition. Particular GAG compositions vary among different tissue types as well as among different developmental and physiological states. Hence, to achieve the proper functional characteristics and obtain reproducible experimental results, it is critical that GAG samples are available with consistent and defined compositions. Unfortunately, most commercially available GAG samples are from animal sources with high inherent variability and are not well characterized. The potential for contamination or adventitious agents makes the animal-derived GAG samples even less desirable for human therapeutics. This STTR Phase I project proposes to develop genetically engineered Chinese Hamster Ovary (CHO) cells producing GAGs with defined compositions. Culture conditions will be optimized to maintain GAG composition while providing product yields at a level sufficient for commercialization. A series of engineered cell lines will be produced in order to prepare GAGs with different compositions that model the different compositions and different functional characteristics found in nature. GAG samples will be characterized both structurally and functionally to provide profiles for each type of sample, which will be reliable and reproducible since the GAGs are prepared from cells grown under controlled conditions. These samples will be a valuable resource for researchers in a number of biological and medical fields. It is thought that the high quality will command premium prices. However, to help reduce costs and promote commercialization, this proposal will test growth conditions in bioreactors to increase the capacity and efficiency of production. Research using these high quality GAG samples will provide proof-of-principal for the use these GAGs in a variety of important therapeutic applications such as oncology, lipid metabolism, tissue regeneration, and hematology.