Functional impact of human collagen α2(XI) gene polymorphism in pathogenesis of ossification of the posterior longitudinal ligament of the spine
Functional impact of human collagen α2(XI) gene polymorphism in pathogenesis of ossification of the posterior longitudinal ligament of the spine
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DOI：
10.1359/jbmr.2001.16.5.948
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发表时间：
2001-05-01
影响因子：
6.2
通讯作者：
Inoue, I
中科院分区：
文献类型：
--
作者：
Maeda, S;Ishidou, Y;Inoue, I
在日本，脊柱后纵韧带骨化（OPLL）是脊髓病的主要原因。在早期的研究中，我们提供了人类胶原蛋白α 2（XI）基因（COL11A2）明显差异的遗传连锁和等位基因关联证据，这些差异可能构成OPLL的遗传易感性。((1))在本研究中，观察到内含子6多态性[内含子6（-4A）]与非OPLL有很强的等位基因关联（p = 0.0003），其中内含子6（-4A）等位基因在非OPEL受试者中比在OPLL患者中更常见。此外，在外显子6 [exon 6（+28A）]上发现一个新的多态性与内含子6（-4A）连锁不平衡。通过比较在棘间韧带培养细胞中通过逆转录聚合酶链反应（RT-PCR）分析的信使RNA（mRNA）剪接的差异和体外外显子捕获研究，分析多态性的功能影响。内含子6（-4A）等位基因导致外显子6跳跃和外显子7保留，而外显子6（+28A）等位基因与mRNA剪接的改变无关。在未分化的成骨细胞（Ob）和非OPLL受试者的后纵韧带细胞中观察到相似的mRNA种类。包含外显子6 - 8的区域是一个酸性亚结构域，可能暴露于表面，可以与细胞外基质的分子相互作用。因此，保留外显子7并去除内含子6（-4A）中观察到的外显子6可能在异位骨化过程中起保护作用，因为在未分化的Ob细胞和未骨化的后纵韧带细胞中观察到相同的模式。
Ossification of the posterior longitudinal ligament (OPLL) of the spine is the leading cause of myelopathy in Japan, In earlier studies, we provided genetic linkage and allelic association evidence of distinct differences in the human collagen alpha2(XI) gene (COL11A2) that might constitute inherited predisposition to OPLL.((1)) In the present study, a strong allelic association with non-OPLL (p = 0.0003) was observed with an intron 6 polymorphism [intron 6 (-4A)], in which the intron 6 (-4A) allele is more frequently observed in non-OPEL subjects than in OPLL patients. In addition, a newly identified polymorphism in exon 6 [exon 6 (+28A)] was in linkage disequilibrium with the intron 6 (-4A). The functional impact of the polymorphisms was analyzed by comparing the differences in messenger RNA (mRNA) splicing by reverse-transcription polymerase chain reaction (RT-PCR) analysis in cultured cells from the interspinous ligament and an in vitro exon trapping study. The intron 6 (-4A) allele resulted in skipping exon 6 and retaining ex:on 7, while the exon 6 (+28A) allele was not associated with alteration in mRNA splicing, Similar mRNA species were observed in undifferentiated osteoblast (Ob) cells and in cells from posterior longitudinal ligament of non-OPLL subjects. The region containing exons 6-8 is an acidic subdomain presumably exposed to the surface that could interact with molecules of the extracellular matrix. Accordingly, retaining exon 7 together with removal of exon 6 observed in intron 6 (-4A) could play a protective role in the ectopic ossification process because the same pattern was observed in undifferentiated Ob cells and nonossified posterior longitudinal ligament cells.