Department of Anatomy, School of Biomedical Sciences, University of Otago, P.O. Box 56, Dunedin 9054, New Zealand.
Hum Mol Genet. 2024 Apr 18;33(9):787-801. doi: 10.1093/hmg/ddae011.
Adolescent idiopathic scoliosis (AIS) is the most common form of scoliosis, in which spinal curvature develops in adolescence, and 90% of patients are female. Scoliosis is a debilitating disease that often requires bracing or surgery in severe cases. AIS affects 2%-5.2% of the population; however, the biological origin of the disease remains poorly understood. In this study, we aimed to determine the function of a highly conserved genomic region previously linked to AIS using a mouse model generated by CRISPR-CAS9 gene editing to knockout this area of the genome to understand better its contribution to AIS, which we named AIS_CRMΔ. We also investigated the upstream factors that regulate the activity of this enhancer in vivo, whether the spatial expression of the LBX1 protein would change with the loss of AIS-CRM function, and whether any phenotype would arise after deletion of this region. We found a significant increase in mRNA expression in the developing neural tube at E10.5, and E12.5, for not only Lbx1 but also other neighboring genes. Adult knockout mice showed vertebral rotation and proprioceptive deficits, also observed in human AIS patients. In conclusion, our study sheds light on the elusive biological origins of AIS, by targeting and investigating a highly conserved genomic region linked to AIS in humans. These findings provide valuable insights into the function of the investigated region and contribute to our understanding of the underlying causes of this debilitating disease.
青少年特发性脊柱侧凸（AIS）是最常见的脊柱侧凸类型，其脊柱在青春期发生弯曲，90%的患者为女性。脊柱侧凸是一种使人衰弱的疾病，在严重的情况下，通常需要支架或手术。AIS 影响人群的 2%-5.2%；然而，这种疾病的生物学起源仍知之甚少。在这项研究中，我们旨在使用 CRISPR-CAS9 基因编辑技术生成的小鼠模型来确定以前与 AIS 相关的高度保守基因组区域的功能，该区域的基因组被敲除，以更好地了解其对 AIS 的贡献，我们将其命名为 AIS_CRMΔ。我们还研究了调节该增强子在体内活性的上游因子，LBX1 蛋白的空间表达是否会随着 AIS-CRM 功能丧失而改变，以及删除该区域后是否会出现任何表型。我们发现，在 E10.5 和 E12.5 时，不仅 Lbx1，而且其他相邻基因的发育中的神经管中的 mRNA 表达都显著增加。成年敲除小鼠表现出椎体旋转和本体感觉缺陷，这也在人类 AIS 患者中观察到。总之，我们的研究通过针对和研究与人类 AIS 相关的高度保守基因组区域，揭示了 AIS 难以捉摸的生物学起源。这些发现为研究区域的功能提供了有价值的见解，并有助于我们理解这种使人衰弱的疾病的根本原因。
