Regeneration of hyaline cartilage in situ with bone marrow mesenchymal stem cells

骨髓间充质干细胞原位再生透明软骨

基本信息

批准号：
10058203
负责人：
CONG-QIU CHU
金额：
--
依托单位：
PORTLAND VA MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-01-01 至 2020-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10058203
关键词：
Adipose tissue Adult Affect Animal Model Animals Antibodies Autologous Biological Assay Biomechanics Bone Marrow Cartilage Cartilage injury Cell Differentiation process Cells Characteristics Charge Chimeric Proteins Chondrocytes Chondrogenesis Chronic Clinical Trials Collagen Collagen Type I Collagen Type II Contralateral Data Defect Degenerative polyarthritis Delayed Hypersensitivity Development Ensure Failure Family Fibrocartilages Fluorescence Future Genes Histologic Histology Human Hyaline Cartilage Hybrids Immune response Immunohistochemistry Impairment In Situ In Vitro Knee Knowledge Lead Lesion Life Measures Mediating Membrane Mesenchymal Differentiation Mesenchymal Stem Cells Methods Modeling Molecular Natural regeneration Oryctolagus cuniculus Outcome Pathology Pathway interactions Phenotype Prevention Procedures Process Production Property Proteins Reaction Regulation Repression Research SOX9 protein Serological Serum Signaling Protein Site Source Synovial Membrane Synovial joint T cell response T-Lymphocyte Techniques Tertiary Protein Structure Testing Therapeutic Effect Time Tissues Transplantation Treatment Efficacy United States Variant aggrecan articular cartilage cartilage development cartilage regeneration cartilage repair clinical development cost design experimental study functional improvement immunogenicity improved improved outcome in vivo in vivo regeneration injury and repair lymphocyte proliferation migration minimally invasive novel therapeutics prevent public health relevance recruit repaired self-renewal stem cell differentiation stem cell proliferation subchondral bone success therapy design tissue repair transcription factor

项目摘要

DESCRIPTION (provided by applicant): Cartilage injury is common which can lead to progression of osteoarthritis (OA) which is characterized by progressive breakdown of articular cartilage, and ultimately leads to functional failure of synovial joints. Regeneration of cartilage has been an attractive approach to cartilage repair and treatment of OA. However, autologous chondrocyte transplantation suffers from insufficient cell supply, new damage to the donor sites and chondrocytes dedifferentiation during in vitro expansion. Mesenchymal stem cells (MSC) which reside in the bone marrow and many adult tissues are capable of self-renewal and differentiation into chondrocytes and are a potential source for cartilage in situ regeneration. Microfracture induces migration of bone marrow MSC to the site of cartilage defect and promotes fibrocartilage production. However, fibrocartilage is non-durable and functionally inadequate in the long-term. SOX9 is a transcription factor belonging to the SOX (Sry-type HMG box) family and has been identified as a "master regulator" of the chondrocyte phenotype. We have generated a cell penetrating, super positively charged SOX9 fusion protein (scSOX9). In vitro studies have demonstrated that scSOX9 protein induced bone marrow derived MSC proliferation and differentiation into chondrocytes. This was evident by decreased production of collagen type I and type X but increased collagen type II and aggrecan, which are characteristics of articular chondrocytes. In a 21 day culture, the initial one time addition of scSOX9 was sufficient to differentiate MSC into chondrocytes and maintain the chondrocyte phenotype. Preliminary in vivo data demonstrated scSOX9 significantly improved the outcome of microfracture in cartilage repair in short-term. In this proposal, we aim to promote cartilage in situ regeneration by stimulating bone marrow MSC in a rabbit model of cartilage defect. Experiments are designed to administer scSOX9 protein at the site of microfracture of the defective cartilage. Administration of scSOX9 will be carried out by either direct delivery with a bioscaffold, collagen membrane at the site of microfracture. Regeneration of cartilage will be assessed at 12 and 24 weeks using histology and immunohistochemistry for characters of hyaline cartilage by examination of content of aggrecan and collagen type I, II and X and osteocalcine. The biomechanical properties of the repaired cartilage will also be evaluated. Immune response to scSOX9 will be assessed by measuring serum antibodies and T cells reaction to scSOX9. Efficacy of re-administration of scSOX9 with microfracture will be assessed in animals who received scSOX9 previously in contralateral knee. Molecular mechanism for which scSOX9 mediated hyaline cartilage repair will be investigated. The research plan takes advantage of the minimal invasive, simple and low cost microfracture to attract bone marrow MSC and pro-chondrogenic property of scSOX9 to regenerate articular cartilage in situ for therapy of cartilage defect. The success of this project will provide invaluable information for design of a clinical trial using the same techniques for therapy of cartilage lesions and prevention of progression to OA.

描述（由申请人提供）：软骨损伤很常见，可导致骨关节炎（OA）的进展，其特征是关节软骨进行性破坏，并最终导致滑膜关节的功能衰竭，软骨再生已成为软骨修复和治疗 OA 的一种有吸引力的方法。然而，自体软骨细胞移植存在细胞供应不足、供体部位新损伤以及体外扩增过程中软骨细胞去分化等问题。骨髓和许多成体组织中的 MSC 能够自我更新并分化为软骨细胞，是软骨原位再生的潜在来源，微骨折可诱导骨髓 MSC 迁移到软骨缺损部位并促进纤维软骨的产生。 SOX9 是一种属于 SOX（Sry 型 HMG 盒）家族的转录因子，已被确定为软骨细胞表型的“主调节器” 我们已经生成了一种细胞穿透性超正电荷 SOX9 融合蛋白 (scSOX9)。体外研究表明，scSOX9 蛋白可诱导骨髓来源的 MSC 增殖并分化为软骨细胞。 I 型和 X 型胶原蛋白的产生，但 II 型胶原蛋白和聚集蛋白聚糖的增加，这是关节软骨细胞的特征。在 21 天的培养中，最初一次性添加scSOX9 足以将 MSC 分化为软骨细胞并维持软骨细胞表型。初步体内数据表明，scSOX9 在短期内显着改善软骨修复中的微骨折的结果。在本提案中，我们的目标是通过刺激骨髓来促进软骨原位再生。软骨缺损兔模型中的 MSC 实验设计为在有缺陷的软骨的微骨折部位施用 scSOX9 蛋白。 scSOX9 将通过生物支架直接递送进行，微骨折部位的胶原膜将在 12 周和 24 周时通过检查聚集蛋白聚糖和胶原类型的含量，使用组织学和免疫组织化学评估透明软骨的特征。 I、II 和 X 以及骨钙素还将评估修复软骨的生物力学特性。通过测量血清抗体和 T 细胞对 scSOX9 的反应，将在先前接受 scSOX9 的对侧膝关节的动物中评估重新施用 scSOX9 的功效。将研究 scSOX9 介导的透明软骨修复的分子机制。利用微创、简单、低成本的微骨折吸引骨髓间充质干细胞和scSOX9的促软骨形成特性来再生关节软骨原位治疗软骨缺损该项目的成功。将为使用相同技术治疗软骨损伤和预防进展为 OA 的临床试验设计提供宝贵的信息。