Variation in the Kozak sequence of WNT16 results in an increased translation and is associated with osteoporosis related parameters
Variation in the Kozak sequence of WNT16 results in an increased translation and is associated with osteoporosis related parameters
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DOI：
10.1016/j.bone.2013.10.022
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发表时间：
2014-02-01
期刊：
影响因子：
4.1
通讯作者：
Van Hul, Wim
中科院分区：
文献类型：
--
作者：
Hendrickx, Gretl;Boudin, Eveline;Van Hul, Wim
WNT 16在骨代谢调节中的重要性最近被几个全基因组关联研究和Wnt 16（Wnt 16（-/-））敲除小鼠模型证实。因此，本研究的目的是复制并进一步阐明WNT 16中常见遗传变异对骨质疏松症相关参数的影响。为此，我们在来自Odense雄激素研究（OAS）的健康高加索男性群体中进行了WNT 16候选基因关联研究。使用HapMap，选择5个tagSNPs和1个多标记测试进行基因分型，以覆盖WNT 16中和周围的大多数常见遗传变异（MAF > 5%）。这项研究证实了先前报道的rs3801387和rs 2707466与几个部位骨密度（BMD）的相关性。此外，我们还证明了rs 2908007与年轻人、老年人和完全OAS人群中几个部位的BMD密切相关。观察到的这三种相关SNP对各自表型的影响是相当的，我们可以得出结论，次要等位基因的存在导致BMD增加。此外，我们对从年轻OAS队列中选择的两个队列进行了WNT 16的重新测序，基于其极端BMD值。在此基础上，选择rs 55710688用于体外翻译实验，因为它位于WNT 16 a的Kozak序列中。我们观察到翻译效率增加，因此Kozak序列的WNT 16 a量更高，这在高BMD队列中显著更普遍。该观察结果与Wnt 16（-/-）小鼠的结果一致。最后，进行了WNT荧光素酶报告基因测定，结果显示Wnt 16没有激活β-连环蛋白依赖性途径。我们确实检测到Wnt 16对WNT 1激活这一经典WNT途径的剂量依赖性抑制作用。因此，WNT 16的翻译增加可导致WNT 16对经典WNT信号传导的抑制作用增加。这种说法与经典WNT信号传导对骨形成的已知激活作用相反，并表明通过非经典WNT信号传导对骨代谢的刺激作用。需要更多的研究不仅要证实这一假设，而且要进一步阐明非经典WNT通路在骨代谢中的作用以及不同WNT信号通路之间相互作用的一般机制。(C)2013 Elsevier Inc. All rights reserved.
The importance of WNT16 in the regulation of bone metabolism was recently confirmed by several genome-wide association studies and by a Wnt16 (Wnt16(-/-)) knockout mouse model. The aim of this study was thus to replicate and further elucidate the effect of common genetic variation in WNT16 on osteoporosis related parameters. Hereto, we performed a WNT16 candidate gene association study in a population of healthy Caucasian men from the Odense Androgen Study (OAS). Using HapMap, five tagSNPs and one multimarker test were selected for genotyping to cover most of the common genetic variation in and around WNT16 (MAF > 5%). This study confirmed previously reported associations for rs3801387 and rs2707466 with bone mineral density (BMD) at several sites. Furthermore, we additionally demonstrated that rs2908007 is strongly associated with BMD at several sites in the young, elderly and complete OAS population. The observed effect of these three associated SNPs on the respective phenotypes is comparable and we can conclude that the presence of the minor allele results in an increase in BMD. Additionally, we performed re-sequencing of WNT16 on two cohorts selected from the young OAS cohort, based on their extreme BMD values. On this basis, rs55710688 was selected for an in vitro translation experiment since it is located in the Kozak sequence of WNT16a. We observed an increased translation efficiency and thus a higher amount of WNT16a for the Kozak sequence that was significantly more prevalent in the high BMD cohort. This observation is in line with the results of the Wnt16(-/-) mice. Finally, a WNT luciferase reporter assay was performed and showed no activation of the beta-catenin dependent pathway by Wnt16. We did detect a dose-dependent inhibitory effect of Wnt16 on WNT1 activation of this canonical WNT pathway. Increased translation of WNT16 can thus lead to an increased inhibitory action of WNT16 on canonical WNT signaling. This statement is in contrast with the known activating effect of canonical WNT signaling on bone formation and suggests a stimulatory effect on bone metabolism via noncanonical WNT signaling. More research is required to not only confirm this hypothesis, but also to further elucidate the role of non-canonical WNT pathways in bone metabolism and the general mechanisms of interplay between the different WNT signaling pathways. (C) 2013 Elsevier Inc. All rights reserved.