Improved cartilage regeneration through the selection and use of highly chondrogenic subpopulations of bone marrow mesenchymal stem cells

通过选择和使用骨髓间充质干细胞的高软骨形成亚群来改善软骨再生

• 批准号: MR/K015648/1
• 负责人: Anthony Hollander
• 金额: $ 87.16万
• 依托单位: University of Bristol
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2013
• 资助国家: 英国
• 起止时间: 2013 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=MR%2FK015648%2F1
• 关键词: Improved; cartilage; regeneration; through; selection

The objective of this grant is to develop a reliable and predictable stem cell therapy for the treatment of diseases of cartilage. Mesenchymal stem cells (MSCs) from bone marrow can be used to generate chondrocytes (cartilage cells) for direct implantation or for use in tissue engineering protocols that can be used to create new cartilage in the laboratory. However there is a marked variation in the outcome of cartilage formation when using cells from one patient to another. We have shown that there is also marked variation in cartilage formation when using different clonal populations of MSCs from the same patient. By selecting the most chondrogenic and least chondrogenic of these clones we have been able to undertake a comparison of all the genes expressed by MSCs that are more or less able to form cartilage. Through analysis of differentially expressed genes we have identified one that can be used as a cell surface marker of the most chondrogenic MSCs. Its increased level of expression on undifferentiated MSC clones is associated with a higher volume and quality of engineered cartilage after chondrogenic differentiation and tissue engineering on three-dimensional scaffolds. Separation of MSCs into those expressing the marker and those not expressing the marker on their cell surface allows comparison of the chondrogenic capacity of each subpopulation. In this way we have shown that the subpopulation expressing the marker is clearly more chondrogenic (ie better able to form cartilage) than the subpopulation that does not express the marker. Furthermore, measurement of specific collagens has provided data suggesting that the increased cartilage formation is not associated with an increased risk of calcification of the new cartilage (this is usually an inherent problem when using bone marrow stem cells to make cartilage). These novel observations give us the unique opportunity to develop a methodology for the production of MSCs that are predictable in their improved capacity to form cartilage.This selected population of cells could be used in cartilage tissue engineering procedures or they could be implanted directly into cartilage lesions without the need for in vitro tissue formation. We now wish to develop robust methods for isolation of stem cells expressing the new marker so that we can develop a cell production method that in a Good Manufacturing Practice (GMP) setting that will be required for regulatory reasons if the cells are to be used to treat patients. We then wish to test this population in a sheep model of articular cartilage damage. We also wish to investigate the capacity of cells expressing the marker to suppress immune responses. Bone marrow mesenchymal stem cells are normally able to suppress some aspects of the immune response but we do not yet know if this property is retained in those cells expressing the new marker. This information may be critical in deciding whether in future to develop strategies for cartilage repair based on the patietns own cells or donated cells. Finally we wish to establish if the marker is a passive molecule that happens to be related to cartilage formation or if it plays a mechanistic role in the way stem cells form cartilage.

这笔赠款的目的是开发一种可靠且可预测的干细胞疗法来治疗软骨疾病。来自骨髓的间充质干细胞 (MSC) 可用于生成软骨细胞（软骨细胞），用于直接植入或用于可在实验室中创建新软骨的组织工程方案。然而，当使用一位患者的细胞与另一位患者的细胞时，软骨形成的结果存在显着差异。我们已经证明，当使用来自同一患者的不同克隆群的 MSC 时，软骨形成也存在显着差异。通过选择这些克隆中软骨形成最强和软骨形成最弱的克隆，我们已经能够对或多或少能够形成软骨的 MSC 表达的所有基因进行比较。通过对差异表达基因的分析，我们已经鉴定出一种可用作最具软骨形成能力的 MSC 的细胞表面标记物。其在未分化的 MSC 克隆上表达水平的增加与软骨分化和三维支架上的组织工程后工程化软骨的更高体积和质量相关。将 MSC 分为细胞表面表达标记的 MSC 和不表达标记的 MSC，以便比较每个亚群的软骨形成能力。通过这种方式，我们已经表明表达该标记的亚群明显比不表达该标记的亚群更具软骨形成性（即能够更好地形成软骨）。此外，特定胶原蛋白的测量提供的数据表明，软骨形成的增加与新软骨钙化风险的增加无关（这通常是使用骨髓干细胞制造软骨时的固有问题）。这些新的观察结果为我们提供了独特的机会来开发一种生产间充质干细胞的方法，该方法可预测其形成软骨的能力的提高。这种选定的细胞群可以用于软骨组织工程程序，也可以直接植入软骨病变中无需体外组织形成。我们现在希望开发强大的方法来分离表达新标记的干细胞，以便我们可以开发一种在良好生产规范（GMP）环境中的细胞生产方法，如果细胞要用于治疗病人。然后我们希望在绵羊关节软骨损伤模型中测试这一群体。我们还希望研究表达标记物的细胞抑制免疫反应的能力。骨髓间充质干细胞通常能够抑制免疫反应的某些方面，但我们尚不清楚这种特性是否保留在表达新标记物的细胞中。这些信息对于决定未来是否开发基于患者自身细胞或捐赠细胞的软骨修复策略可能至关重要。最后，我们希望确定该标记物是否是一种与软骨形成相关的被动分子，或者它是否在干细胞形成软骨的过程中发挥机械作用。

项目成果

Human fetal and adult bone marrow-derived mesenchymal stem cells use different signaling pathways for the initiation of chondrogenesis.

人类胎儿和成人骨髓间充质干细胞使用不同的信号通路来启动软骨形成。

• DOI: 10.1089/scd.2013.0301
• 发表时间: 2014-03-01
• 期刊: Stem cells and development
• 影响因子: 4
• 作者: Kyla Brady;S. Dickinson;P. Guillot;J. Polak;A. Blom;W. Kafienah;A. Holl;er;er
• 通讯作者: er

The Wnt5a Receptor, Receptor Tyrosine Kinase-Like Orphan Receptor 2, Is a Predictive Cell Surface Marker of Human Mesenchymal Stem Cells with an Enhanced Capacity for Chondrogenic Differentiation.

Wnt5a 受体（受体酪氨酸激酶样孤儿受体 2）是人类间充质干细胞的预测性细胞表面标记，具有增强的软骨分化能力。

• DOI: http://dx.10.1002/stem.2691
• 发表时间: 2017
• 期刊: Stem cells (Dayton, Ohio)
• 作者: Dickinson SC