Elucidation of the role of Creb5 in promoting the formation and patterning of the intervertebral disc

项目摘要/摘要。 脊柱疾病很常见，包括椎间盘退变、小关节综合症和脊柱侧弯。 疾病。椎间盘退变和小关节综合症以及脊柱侧弯都是常见的疾病。 与/或可能导致慢性下腰痛，这是非常常见的，导致更多的全球残疾 任何其他情况。这个项目的广泛、长期的目标是发展对 细胞身份的规范和调节形成和图案化的调节网络 椎间盘在脊柱功能中起着关键作用，容易患上发育性疾病， 退化性疾病和损伤。然而，像关节软骨受损后，椎间盘细胞不会 修复/再生良好。因此，确定转录因子和/或信号分子是至关重要的 这决定了不同类型的细胞(包括干细胞/祖细胞)在椎间盘中形成不同的 治疗脊柱疾病的策略。我最近的发现表明，转录因子Creb5是 在IVD形成前在椎间充质细胞中特异表达；随后在 纤维环内的所有细胞，包括纤维内环和外环 纤维组织(OAF)在IVD形成后表达，在小关节也有表达。最重要的是，我发现 在缺乏Creb5功能的基因工程小鼠中，纤维环细胞的形态发生了显着变化； Prg4/Luusin和WNT拮抗剂Wif1(Wnt抑制因子1)的表达都显著增加 在这些小鼠的椎间盘中减少。因此，我的发现表明，维持Creb5是必要的 椎间盘的适当形态，并对驱动Prg4/Luxin和Prg4/Luxin的表达至关重要 Wif1在椎间盘内。与Creb5在IVD中的重要性一致，这是一种全基因组的关联 研究发现，Creb5基因是日本青少年特发性脊柱侧凸的易感基因。一个更新的 全基因组关联研究确定Creb5是与退行性变手术相关的易感基因 肩袖疾病，最常见的原因是另一个纤维结缔组织的进行性磨损 (肌腱)。由于Creb5在椎间充质(IM)中特异表达，从而导致 在间盘、小关节和纤维环的所有细胞中，我推测creb5在 并建议进一步研究Creb5在脊柱发育中的作用。这个项目将 确定Creb5在脊柱发育期和成人期的表达模式；将描绘出 表达Creb5的细胞在脊柱发育过程中的去向；并测试Creb5HA-CreERt2小鼠能否成年 以所有成熟的纤维环细胞为靶点的阶段性遗传工具和时间控制。该项目还将确定 Creb5调控的基因和染色质调控元件都能将Creb5结合在椎间盘中。

Project Summary/Abstract. Spine diseases, including intervertebral disc degeneration, facet joint syndrome, and scoliosis, are common diseases. Both intervertebral disc degeneration and facet joint syndrome, as well as scoliosis, are frequently associated with/or may lead to chronic low back pain, which is very common, causing more global disability than any other condition. The broad, long-term goal of this project is to develop a comprehensive understanding of both the specification of the cell identity and the regulatory network that regulates the formation and patterning of the intervertebral disc, which plays a pivotal role in spinal function and is subject to developmental disease, degenerative disease, and injury. However, like articular cartilage after damage, intervertebral disc cells do not repair/regenerate well. Thus, it’s critical to identify both the transcription factors and/or the signaling molecules that determine different cell types (including stem/progenitors) in the intervertebral disc to develop different strategies to treat spine disease. My recent findings have indicated that the transcription factor Creb5 is specifically expressed in intervertebral mesenchymal cells before IVD formation; is subsequently expressed in all the cells in the annulus fibrosus, including both the inner annulus fibrosus (IAF) and the outer annulus fibrosus (OAF) after IVD formation; and is also expressed in facet joints. Most significantly, I have found that the morphology of the annulus fibrosus cells is significantly altered in mice engineered to lack Creb5 function; and that expression of both Prg4/lubricin and the WNT antagonist Wif1 (Wnt inhibitor factor 1) are dramatically decreased in the intervertebral discs of these mice. Thus, my findings indicate that Creb5 is necessary to maintain the proper morphology of the intervertebral disc, and is critical to drive expression of both Prg4/lubricin and Wif1 in the intervertebral disc. Consistent with the importance of Creb5 in the IVD, a genome-wide association study identified Creb5 as a susceptibility locus for adolescent idiopathic scoliosis in Japan. A more recent genome-wide association study identified Creb5 as a susceptibility locus associated with surgery for degenerative rotator cuff disease, which is most often caused by progressive wear and tear of another fibrous connective tissue (tendon). As Creb5 is specifically expressed in the intervertebral mesenchymal (IM) that gives rise to the intervertebral disc, in facet joints, and in all the cells of annulus fibrosus, I hypothesize that Creb5 plays a critical role in the spine and propose to further investigate the role of Creb5 in spine development. This project will determine the expression pattern of Creb5 during spine development stage and adult stage; will delineate the fate of Creb5-expressing cells during spine development; and test whether Creb5HA-CreERt2 mice can be an adult stage genetic tool with temporal control to target all mature annulus fibrosus cells. This project will also identify both Creb5 regulated genes and chromatin regulatory elements that bind Creb5 in the intervertebral disc.