Conditional ablation of Meningioma-1 in osteoblasts

描述(申请人提供)：维生素D受体(VDR)，其必要的激素配体1，25-二羟基维生素D3(1，25(OH)2D3)，以及许多核受体共激活因子和辅阻遏子，通过增加肠道钙吸收和直接影响成骨细胞的增殖、分化和功能，协调控制选定基因和基因网络的表达，从而影响骨骼。脑膜瘤-1(MN1)是VDR和1，25(OH)2D3在成骨细胞中调控的靶基因之一。MN1是VDR和其他核受体的共激活剂，它是VDR完全配体诱导的活性所必需的。MN1缺失的小鼠模型的表型表明了MN1在骨骼生物学中的重要意义，该模型在膜内成骨中显示了颅骨异常。体外数据支持这种新的因子在成骨细胞和骨骼生物学中的重要作用。例如，MN1在包括原代成骨细胞在内的许多成骨细胞系中表达，在成骨细胞分化过程中其表达增加，在共培养研究中，MN1的去除显著影响成骨细胞的形态、成骨细胞的增殖、成骨细胞的分化、1，25(OH)2D3激活的基因表达以及1，25(OH)2D3刺激的破骨细胞形成。然而，这些体外方法并没有解决MN1在出生后骨骼中的生理相关性。由于全球MN1KO模型出生后的致命性，MN1生物学在骨骼中的这个关键位置仍然没有得到解决。在目前的提案中，我们提供了关于开发解决这一问题的新鼠标模型的初步数据。我们的实验室利用Col？1(I)-cre缺失株和条件等位基因相结合，开发了一种针对成骨细胞的小鼠MN1基因的消融小鼠，在条件等位基因中，小鼠MN1基因的外显子1两侧是loxP位点。Col？1(I)-cre+MN1fl/fl小鼠在20周龄时表现出性别选择性和骨室选择性的矿化不足表型。这些缺陷让人想起在人类和啮齿动物模型中观察到的年轻人在正常衰老过程中早期骨丢失的情况。这些新数据表明，MN1参与了更多的全球骨骼重塑，独立于VDR和维生素D内分泌系统。这一最新模型有可能揭示幼年成年哺乳动物早期骨小梁丢失的潜在机制和生物学机制。因此，这项建议的总体单一研究目标是描述这一新模型的详细骨骼表型，并确立MN1在出生后骨骼生理学中的意义。

DESCRIPTION (provided by applicant): The vitamin D receptor (VDR), its requisite hormonal ligand, 1,25-dihydroxyvitamin D3 (1,25(OH)2D3), as well as a host of nuclear receptor coactivators and corepressors coordinately control the expression of select genes and gene networks to impact bone by increasing intestinal calcium absorption and by directly affecting osteoblast proliferation, differentiation, and function. Meningioma-1 (MN1) is one such target gene that VDR and 1,25(OH)2D3 regulate in osteoblasts. MN1 is a coactivator of VDR and other nuclear receptors and it is needed for full ligand-induced activity of the VDR. The significance of MN1 in skeletal biology is suggested by the phenotype of the MN1 null mouse model which displays cranial skeletal anomalies in intramembranous ossification. In vitro data support an important role for this novel factor in the osteoblast and in skeletal biology. For example, MN1 is expressed in numerous osteoblastic cell lines including primary osteoblasts, its expression is increased during osteoblast differentiation, MN1 ablation dramatically impacts osteoblast morphology, osteoblast proliferation, osteoblast differentiation, 1,25(OH)2D3-activated gene expression, and 1,25(OH)2D3-stimulated osteoclastogenesis in co-culture studies. However, these in vitro approaches do not address the physiological relevance of MN1 in the postnatal skeleton. Due to postnatal lethality of the global MN1KO model, this critical poin of MN1 biology in the skeleton has remained unaddressed. In this current proposal, we present preliminary data on the development of a new mouse model that addresses this issue. Our laboratory developed a mouse with osteoblast-targeted ablation of the murine MN1 gene using the Col¿1(I)-cre deletor strain combined with a conditional allele in which exon 1 of the murine MN1 gene is flanked by loxP sites. The Col¿1(I)-cre+MN1fl/fl mice displayed a gender-selective and compartment-selective undermineralized skeletal phenotype at 20 weeks of age. These defects are reminiscent of those observed in early bone loss in young adults during the normal aging process in human and rodent models. These new data implicate MN1 in more global skeletal remodeling independent of VDR and the vitamin D endocrine system. This newest model has the potential to uncover new insight into potential mechanisms and biologies involved in early trabecular bone loss in young, adult mammals. Thus, the overall single research goal of this proposal is to characterize the detailed skeletal phenotype of this new model and establish the significance of MN1 in postnatal skeletal physiology.