Developing a stable cell line expressing recombinant sclerostin

开发表达重组硬化素的稳定细胞系

基本信息

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
项目类别：
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

项目摘要

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 摘要 2 颞下颌关节 (TMJ) 是一个复杂的关节系统，对于牙齿咬合至关重要， 3 咀嚼、呼吸和言语颞下颌关节由肌肉网络组成， 4 条韧带，以及纤维软骨盘和髁突。 5 是涉及颞下颌关节和相关疾病的一组复杂、异质的病理学 6 退行性类型的 TMD，包括 TMJ 骨关节炎 (OA)，使人衰弱， 7 影响生活质量的 TMD 每年影响超过 1000 万美国人。 8 根据 NIDCR，目前的 TMJ OA 治疗通常是两倍，涉及其中之一。 9 疼痛管理或侵入性手术，例如失败率很高的全关节置换术。 10 再生医学有望恢复因疾病而丧失的功能组织。 11 因此，利用常驻干细胞的再生能力来修复颞下颌关节可能图12代表由NIH赞助的基于干细胞的微创治疗TMJ OA。 13小企业资助，我们进一步开发了StemGel®，一种缓释制剂 14 将硬化蛋白封装在透明质酸 (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 个型号。 17 StemGel® 的活性成分硬化素是一种含有 213 个氨基酸的糖基化蛋白质。 18 需要哺乳动物表达系统来忠实地产生翻译后表达 19 修饰是治疗性蛋白的关键拐点。 20 细胞基质的特性以及与细胞基质相关的事件。 21会影响蛋白质的质量和安全性，更进一步，对蛋白质各方面进行有效的质量控制 22 处理细胞底物该提案的目标是开发主细胞库。 23 (MCB)/表达硬化素的稳定细胞系可轻松转移至合同 24 用于大规模生产 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。