Developing a stable cell line expressing recombinant sclerostin

开发表达重组硬化素的稳定细胞系

• 批准号: 10385037
• 负责人: Mo Chen
• 金额: $ 29万
• 依托单位: WNT SCIENTIFIC, LLC
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-20 至 2024-09-16
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10385037
• 关键词: Affect; American; Amino Acids; Award; Biological; Biological Products; Biological Sciences; Biology; Biotechnology; Businesses; Cartilage; Cell Line; Cell Therapy; Cells; Chinese Hamster Ovary Cell; Complex; Contracts; Data; Dental Occlusion; Derivation procedure; Development; Disease; Drug Kinetics; Encapsulated; Event; Failure; Fibrocartilages; Formulation; Goals; Grant; Guidelines; Human; Human Cell Line; Hyaluronic Acid; Injections; Innovative Therapy; Intra-Articular Injections; Joints; Lead; Ligaments; Link; Mastication; Miniature Swine; Modeling; Modification; Mus; Muscle; National Institute of Dental and Craniofacial Research; Natural regeneration; New Jersey; Operative Surgical Procedures; Oryctolagus cuniculus; Pain management; Pathology; Phase; Post-Translational Protein Processing; Production; Property; Proteins; Protocols documentation; Quality Control; Quality of life; Recombinants; Recreation; Regenerative Medicine; Replacement Arthroplasty; Research; Respiration; Safety; Signal Transduction; Small Business Technology Transfer Research; Speech; System; Temporomandibular Joint; Temporomandibular Joint Disorders; Temporomandibular joint osteoarthritis; Therapeutic; Therapeutic antibodies; Tissues; United States National Institutes of Health; Universities; Vendor; Work; beta catenin; cell bank; cost; inhibitor; innovation; joint injury; minimally invasive; operation; pre-clinical research; product development; protein purification; regenerative; repaired; safety testing; scale up; stable cell line; stem cell therapy; stem cells; therapeutic protein; treatment strategy

1 Abstract 2 The temporomandibular joint (TMJ) is a complex joint system critical for dental occlusion, 3 mastication, respiration and speech. The TMJ is comprised of a network of muscles, 4 ligaments, and a fibrocartilaginous disc and condyle. Temporomandibular disorders (TMDs) 5 are a complex, heterogeneous group of pathologies involving the TMJ and associated 6 musculature. Degenerative types of TMDs, including TMJ osteoarthritis (OA), are debilitating, 7 compromise the quality of life. TMDs afflict over 10 million Americans at an annual cost of 8 ~$4 billion, per the NIDCR. Current TMJ OA treatments are typically two-fold, involving either 9 pain management or invasive surgeries, such as total joint replacements with high failure rates. 10 Regenerative medicine promises the recreation functional tissue that is lost from diseases. 11 Thus exploiting the regenerative capabilities of resident stem cells to repair TMJ may 12 represent a minimally invasive stem cell based treatment for TMJ OA. Sponsored by NIH 13 small business grant, we further developed StemGel®, a sustained-release formulation by 14 encapsulating sclerostin in hyaluronic acid (HA). Monthly injection of StemGel® efficiently 15 inhibits TMJ OA progression and regenerates cartilage in both rabbit and minipig TMJ OA 16 models. 17 The active ingredient of StemGel®, sclerostin is a glycosylated protein with 213 amino acids. 18 A mammalian expressing system is required to faithfully produce the post-translational 19 modification. Cell line development is a critical inflection point for therapeutic protein 20 manufacturing. The properties of the cell substrate and the events linked to cell substrate 21 can affect protein quality and safety, and further, the effective quality control on all aspects of 22 handling the cell substrate. The goal of this proposal is to develop Master Cell Bank 23 (MCB)/stable cell line expressing sclerostin to be readily transferred to the Contract 24 Development and Manufacturing Organization (CDMO) for massive production of the GMP 25 grade protein for pharmacokinetics and safety tests.

1篇摘要 2颞下颌关节（TMJ）是一个复杂的关节系统， 3咀嚼、呼吸和言语。颞下颌关节由肌肉网络组成， 4韧带，和纤维软骨盘和髁。颞下颌关节紊乱病 5是一个复杂的，异质性的病理组，涉及颞下颌关节和相关的 6肌肉组织。退行性类型的TMD，包括TMJ骨关节炎（OA），是使人衰弱的， 7、降低生活质量。TMD折磨着超过1000万美国人，每年的费用为 80 ~ 40亿美元，根据NIDCR。目前的TMJ OA治疗通常是双重的，包括 9疼痛管理或侵入性手术，如失败率高的全关节置换术。 10再生医学承诺重建因疾病而失去的功能组织。 因此，利用常驻干细胞的再生能力来修复颞下颌关节可能 12代表了一种基于干细胞的微创治疗TMJ OA的方法。由NIH赞助 13小企业赠款，我们进一步开发了StemGel®，一种缓释制剂， 14将硬化蛋白包封在透明质酸（HA）中。每月有效注射StemGel® 15抑制兔和小型猪TMJ OA的进展并再生软骨 16个模特 StemGel®的活性成分sclerostin是一种含有213个氨基酸的糖基化蛋白质。 18需要哺乳动物表达系统忠实地产生翻译后蛋白。 19修改细胞系开发是治疗性蛋白质的关键转折点 20制造业细胞基质的性质和与细胞基质相关的事件 21可以影响蛋白质的质量和安全性，并且进一步地，有效的质量控制的各个方面， 22处理电池基板。本提案的目标是开发主细胞库 23（MCB）/表达sclerostin的稳定细胞系可随时转移至合同 24大规模生产GMP的开发和制造组织（CDMO） 用于药代动力学和安全性测试的25级蛋白质。