N-cadherin is Key to Expression of the Nucleus Pulposus Cell Phenotype under Selective Substrate Culture Conditions.
N-cadherin is Key to Expression of the Nucleus Pulposus Cell Phenotype under Selective Substrate Culture Conditions.
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DOI：
10.1038/srep28038
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发表时间：
2016-06-13
影响因子：
4.6
通讯作者：
Setton LA
中科院分区：
文献类型：
--
作者：
Hwang PY;Jing L;Chen J;Lim FL;Tang R;Choi H;Cheung KM;Risbud MV;Gersbach CA;Guilak F;Leung VY;Setton LA
椎间盘髓核（NP）细胞对合成细胞外基质至关重要，而细胞外基质有助于椎间盘的健康和机械功能。NP细胞由于其脊索起源而具有独特的形态和分子表达模式，并且在体内驻留在N-cadherin （CDH2）阳性的细胞簇中。随着椎间盘退变，NP细胞发生形态和表型变化，包括CDH2表达的丧失和形成细胞簇的能力。在这里，我们研究了CDH2阳性细胞簇在保存健康、生物合成活性NP细胞中的作用。使用层粘胶蛋白功能化的水凝胶系统来模拟天然NP微环境的特征，我们证明只有当NP细胞形成CDH2阳性细胞簇时，NP细胞的表型和形态才会被保留。体外和体内敲低（CRISPRi）或阻断CDH2表达会导致健康NP细胞的丧失。研究结果还表明，在受控微环境条件下，通过促进细胞聚集，可以促进CDH2阴性的退化人NP细胞重新表达CDH2和健康的、幼年的NP基质合成模式。这项工作还确定了CDH2与β-连环蛋白调节信号的相互作用是CDH2介导的细胞相互作用控制NP细胞表型和生物合成以维持健康椎间盘组织的一种机制。
Nucleus pulposus (NP) cells of the intervertebral disc are essential for synthesizing extracellular matrix that contributes to disc health and mechanical function. NP cells have a unique morphology and molecular expression pattern derived from their notochordal origin, and reside in N-cadherin (CDH2) positive cell clusters in vivo. With disc degeneration, NP cells undergo morphologic and phenotypic changes including loss of CDH2 expression and ability to form cell clusters. Here, we investigate the role of CDH2 positive cell clusters in preserving healthy, biosynthetically active NP cells. Using a laminin-functionalized hydrogel system designed to mimic features of the native NP microenvironment, we demonstrate NP cell phenotype and morphology is preserved only when NP cells form CDH2 positive cell clusters. Knockdown (CRISPRi) or blocking CDH2 expression in vitro and in vivo results in loss of a healthy NP cell. Findings also reveal that degenerate human NP cells that are CDH2 negative can be promoted to re-express CDH2 and healthy, juvenile NP matrix synthesis patterns by promoting cell clustering for controlled microenvironment conditions. This work also identifies CDH2 interactions with β-catenin-regulated signaling as one mechanism by which CDH2-mediated cell interactions can control NP cell phenotype and biosynthesis towards maintenance of healthy intervertebral disc tissues.