STTR Phase I: Biomanufacture of Novel Heparan Sulfate Glycosaminoglycans

这个小企业技术转让（STTR）项目的更广泛的影响/商业潜力将是将称为糖胺聚糖（GAG）的新型糖聚合物商业化，用于各种重要的医疗应用。GAG是许多重要的生理和病理生理状况（例如，肿瘤血管生成、血栓形成）;然而，由于缺乏用于聚糖分析的分析技术和缺乏可用于研究的高质量GAG样品，因此对这些可能显著影响人类健康的作用进行详细了解具有挑战性。GAG在细胞信号传导级联中具有精确的功能作用，其可以根据特定的GAG结构组成而变化。具体的GAG组成在不同组织类型之间以及在不同发育和生理状态之间变化。因此，为了实现适当的功能特性并获得可重现的实验结果，必须提供具有一致和确定组成的GAG样品。不幸的是，大多数市售GAG样品来自具有高度固有变异性的动物来源，并且没有得到很好的表征。污染或外源因子的可能性使得动物源性GAG样品更不适合用于人类治疗。该STTR I期项目提出开发基因工程中国卵巢（CHO）细胞，生产具有确定组成的GAG。将优化培养条件，以维持GAG组成，同时提供足以商业化的产品产量。将产生一系列工程化细胞系，以制备具有不同组成的GAG，其模拟自然界中发现的不同组成和不同功能特征。将对GAG样品进行结构和功能表征，以提供每种类型样品的特征，由于GAG是由在受控条件下生长的细胞制备的，因此这些特征将是可靠和可重现的。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。人们认为高质量将要求高价格。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中的生长条件，以提高生产能力和效率。 使用这些高质量GAG样本进行的研究将为这些GAG在各种重要治疗应用（如肿瘤学、脂质代谢、组织再生和血液学）中的使用提供原理证明。

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.