Functional analysis of the osteoarthritis susceptibility-associated GDF5 regulatory polymorphism.
Functional analysis of the osteoarthritis susceptibility-associated GDF5 regulatory polymorphism.
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DOI：
10.1002/art.24616
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发表时间：
2009-07
影响因子：
--
通讯作者：
Loughlin J
中科院分区：
文献类型：
--
作者：
Egli RJ;Southam L;Wilkins JM;Lorenzen I;Pombo-Suarez M;Gonzalez A;Carr A;Chapman K;Loughlin J
GDF5基因5‘非翻译区的单核苷酸多态(SNP)rs143383(T到C)最近被报道与骨关节炎(OA)的易感性有关，在体外和体内观察到与风险相关的T等位基因的低表达。活体研究取自骨性关节炎患者的软骨组织。本研究旨在将这种SNP对GDF5等位基因表达的影响分析扩展到更多的关节组织类型，以寻找与SNP相互作用的顺式和反式因子，并检测新的顺式作用GDF5调控多态。组织样本来自接受髋关节或膝关节置换手术的骨性关节炎患者。提取核酸，使用rs143383和等位基因表达的鉴别和量化分析，测量T和C等位基因GDF5的相对表达量。使用等位基因表达和荧光素酶报告分析将GDF5转录本序列中的其他常见变体作为潜在的调控元件进行询问，并使用凝胶迁移率改变分析来寻找与rs143383结合的反式因子。我们观察到GDF5在所有受试组织中存在一致的等位基因表达失衡，这意味着rs143383对GDF5表达的功能影响是关节范围的。我们发现了位于GDF5基因3‘-非编码区的第二个多态，它影响该基因的等位基因表达，而不依赖于rs143383。最后，我们观察到变形的表皮自体调节因子1与rs143383的2个等位基因的差异结合。这些发现表明，GDF5基因的多态介导的骨性关节炎的易感性并不局限于软骨，强调了疾病累及整个关节的必要性。另外一个顺式作用调控基因的存在突显了这个重要的骨性关节炎易感基因表达调控的复杂性。聋人-1是一种反式作用因子，作为一种潜在的调节GDF5表达的工具，值得进一步研究。
Single-nucleotide polymorphism (SNP) rs143383 (T to C) in the 5′-untranslated region (5′-UTR) of GDF5 has recently been reported to be associated with osteoarthritis (OA) susceptibility, with lower expression of the risk-associated T allele observed in vitro and in vivo. The in vivo studies were performed on cartilage tissue from OA patients. The present study was undertaken to expand the analysis of the effect of this SNP on GDF5 allelic expression to more joint tissue types, to investigate for cis and trans factors that interact with the SNP, and to examine novel cis-acting GDF5 regulatory polymorphisms. Tissue samples were collected from OA patients undergoing joint replacement of the hip or knee. Nucleic acid was extracted, and, using rs143383 and an assay that discriminates and quantifies allelic expression, the relative amount of GDF5 expression from the T and C alleles was measured. Additional common variants in the GDF5 transcript sequence were interrogated as potential regulatory elements using allelic expression and luciferase reporter assays, and electrophoretic mobility shift assays were used to search for trans factors binding to rs143383. We observed a consistent allelic expression imbalance of GDF5 in all tissues tested, implying that the functional effect mediated by rs143383 on GDF5 expression is joint-wide. We identified a second polymorphism, located in the 3′-UTR of GDF5, that influenced allelic expression of the gene independent of rs143383. Finally, we observed differential binding of deformed epidermal autoregulatory factor 1 (DEAF-1) to the 2 alleles of rs143383. These findings show that the OA susceptibility mediated by polymorphism in GDF5 is not restricted to cartilage, emphasizing the need to consider the disease as involving the whole joint. The existence of an additional cis-acting regulatory polymorphism highlights the complexity of the regulation of expression of this important OA susceptibility locus. DEAF-1 is a trans-acting factor that merits further investigation as a potential tool for modulating GDF5 expression.
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影响因子：
5.3
作者：
Hahm, KM;Sum, EYM;Orkin, SH
通讯作者：
Orkin, SH
影响因子：
4
作者：
Hatakeyama, Y;Tuan, RS;Shum, L
通讯作者：
Shum, L
影响因子：
64.5
作者：
Eulalio, Ana;Huntzinger, Eric;Izaurralde, Elisa
通讯作者：
Izaurralde, Elisa
影响因子：
30.8
作者：
Sanna, Serena;Jackson, Anne U.;Mohlke, Karen L.
通讯作者：
Mohlke, Karen L.
影响因子：
3.5
作者：
Wilkins, James M.;Southam, Lorraine;Loughlin, John
通讯作者：
Loughlin, John