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

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

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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）是一个对牙齿阻塞至关重要的复杂关节系统， 3咀嚼，呼吸和言语。 TMJ由肌肉网络组成， 4个韧带，以及纤维状圆盘和con。颞下颌疾病（TMD） 5是涉及TMJ和相关的复杂的，异质的病理组 6肌肉。 TMD的退化类型，包括TMJ骨关节炎（OA），使人衰弱， 7损害生活质量。 TMD遭受了超过1000万美国人的折磨根据NIDCR，8〜40亿美元。当前的TMJ OA治疗通常是两个倍，涉及 9疼痛管理或侵入性手术，例如具有高失败率的总关节置换。 10再生医学有望从疾病中丢失的娱乐功能组织。 11利用居民干细胞修复TMJ的再生能力可能 12代表用于TMJ OA的最小侵入性干细胞处理。由NIH赞助 13小型企业赠款，我们进一步开发了Stemgel®，这是一种持续释放的公式 14透明质酸（HA）中封装硬化蛋白。每月有效地注入Stemgel® 15抑制TMJ OA进展并再生兔子和Minipig TMJ OA的软骨 16个型号。 17STEMGEL®的活性成分，硬化蛋白是一种糖基化的蛋白质，具有213个氨基酸。 18需要忠实地生产哺乳动物表达系统 19修改。细胞系发育是治疗蛋白的关键拐点 20制造。细胞底物的特性以及与细胞基板相关的事件 21可以影响蛋白质质量和安全性，进一步，对所有方面的有效质量控制 22处理细胞底物。该提案的目的是开发大型细胞银行 23（MCB）/表达硬化蛋白的稳定细胞系容易转移到合同中 24大量生产GMP的开发和制造组织（CDMO） 25级蛋白质用于药代动力学和安全测试。