Increased TBX6 gene dosages induce congenital cervical vertebral malformations in humans and mice.
Increased TBX6 gene dosages induce congenital cervical vertebral malformations in humans and mice.
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TBX6基因剂量增加可诱发人和小鼠先天性颈椎畸形
DOI：
10.1136/jmedgenet-2019-106333
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发表时间：
2020-06
影响因子：
4
通讯作者：
Liu, Pengfei
中科院分区：
文献类型：
--
作者：
Ren, Xiaojun;Yang, Nan;Wu, Nan;Xu, Ximing;Chen, Weisheng;Zhang, Ling;Li, Yingping;Du, Ren-Qian;Dong, Shuangshuang;Zhao, Sen;Chen, Shuxia;Jiang, Li-Ping;Wang, Lianlei;Zhang, Jianguo;Wu, Zhihong;Jin, Li;Qiu, Guixing;Lupski, James R.;Shi, Jiangang;Zhang, Feng;Liu, Pengfei
背景先天性椎体畸形（CVMs）表现为椎体形态异常。遗传因素与脑血管畸形的发病机制有关，但其发病机制尚不清楚。我们先前报道了人类16p11.2 BP 4-BP 5缺失及其相关的TBX 6剂量减少导致了CVM。我们的目的是研究16p11.2 BP 4-BP 5的相互重复及其对CVM的潜在遗传贡献。方法和结果通过染色体微阵列分析发现携带16p11.2 BP 4-BP 5重复的患者回顾性分析其椎骨表型。7例重复携带者的脊柱评估显示，4例（57%）表现出脑血管畸形的特征，支持增加TBX 6剂量可诱导脑血管畸形的论点。为了在模型生物体中进行进一步的体内功能研究，我们使用CRISPR-Cas9对小鼠Tbx 6的上游调控区进行了基因组编辑，并获得了Tbx 6表达水平升高的三个小鼠突变等位基因（Tbx 6 up 1至Tbx 6 up3）。荧光素酶报告基因分析显示，Tbx 6 up3等位基因的表达水平为参考（+）等位基因的160%。因此，纯合子Tbx 6 up3/up3小鼠可以在功能上模拟16p11.2 BP 4-BP 5重复的杂合子携带者的TBX 6剂量（约150%，即正常水平的3/2基因剂量）。值得注意的是，60%的Tbx 6 up3/up3小鼠表现出CVM。与我们在人类中的观察一致，在小鼠中由增加的Tbx 6剂量诱导的CVM主要影响颈椎。结论我们在人类和小鼠中的研究结果一致支持增加TBX 6剂量有助于发展宫颈CVM的风险。
Background Congenital vertebral malformations (CVMs) manifest with abnormal vertebral morphology. Genetic factors have been implicated in CVM pathogenesis, but the underlying pathogenic mechanisms remain unclear in most subjects. We previously reported that the human 16p11.2 BP4-BP5 deletion and its associated TBX6 dosage reduction caused CVMs. We aim to investigate the reciprocal 16p11.2 BP4-BP5 duplication and its potential genetic contributions to CVMs. Methods and results Patients who were found to carry the 16p11.2 BP4-BP5 duplication by chromosomal microarray analysis were retrospectively analysed for their vertebral phenotypes. The spinal assessments in seven duplication carriers showed that four (57%) presented characteristics of CVMs, supporting the contention that increased TBX6 dosage could induce CVMs. For further in vivo functional investigation in a model organism, we conducted genome editing of the upstream regulatory region of mouse Tbx6 using CRISPR-Cas9 and obtained three mouse mutant alleles (Tbx6up1 to Tbx6up3 ) with elevated expression levels of Tbx6. Luciferase reporter assays showed that the Tbx6up3 allele presented with the 160% expression level of that observed in the reference (+) allele. Therefore, the homozygous Tbx6up3/up3 mice could functionally mimic the TBX6 dosage of heterozygous carriers of 16p11.2 BP4-BP5 duplication (approximately 150%, ie, 3/2 gene dosage of the normal level). Remarkably, 60% of the Tbx6up3/up3 mice manifested with CVMs. Consistent with our observations in humans, the CVMs induced by increased Tbx6 dosage in mice mainly affected the cervical vertebrae. Conclusion Our findings in humans and mice consistently support that an increased TBX6 dosage contributes to the risk of developing cervical CVMs.
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影响因子：
2
作者：
Al-Kateb, Hussam;Khanna, Geetika;Shinawi, Marwan
通讯作者：
Shinawi, Marwan
影响因子：
4.6
作者：
Krol, Aurelie J.;Roellig, Daniela;Pourquie, Olivier
通讯作者：
Pourquie, Olivier
影响因子：
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作者：
Di Gioia, Silvio Alessandro;Shaaban, Sherin;Engle, Elizabeth C.
通讯作者：
Engle, Elizabeth C.
影响因子：
2.7
作者：
Chapman, DL;Agulnik, I;Papaioannou, VE
通讯作者：
Papaioannou, VE
影响因子：
8.8
作者：
Cheung, SW;Shaw, CA;Beaudet, AL
通讯作者：
Beaudet, AL