Synergistic effects of rare variants of ARHGAP31 and FBLN1 in vitro in terminal transverse limb defects.
Synergistic effects of rare variants of ARHGAP31 and FBLN1 in vitro in terminal transverse limb defects.
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ARHGAP31 和 FBLN1 罕见变体在体外对终末横向肢体缺损的协同作用
DOI：
10.3389/fgene.2022.946854
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发表时间：
2022
影响因子：
3.7
通讯作者：
Li, Chuanzhou
中科院分区：
文献类型：
--
作者：
Tian, Hong;Chu, Fan;Li, Yingjie;Xu, Mengmeng;Li, Wenjiao;Li, Chuanzhou
背景：先天性皮肤发育不全（ACC）和末端横肢缺损（TTLDs）是Adams-Oliver综合征（AOS）最常见的特征。ARHGAP 31是常染色体显性遗传型AOS的致病基因之一，其变异体可能仅引起孤立性TTLD。方法：采用全外显子组测序（WES）和桑格测序技术对TTLD的致病基因进行鉴定。构建表达载体用于在哺乳动物细胞培养物中转染，随后进行生化和功能分析，包括免疫印迹、免疫荧光染色和细胞计数kit-8测定。 结果如下：WES和桑格测序结果提示先证者分别从其无症状的父母之一遗传了罕见的ARHGAP 31变异[c.2623G > A（p.Glu875Lys）]和罕见的FBLN 1变异[c.1649G > A（p.Arg550His）]。鉴于FBLN 1变异也与并指畸形有关，我们怀疑这两个基因共同促成了TTLD表型，并在体外探索了它们可能的作用。突变体FBLN 1表现出由于蛋白质稳定性受损而导致的表达降低，而ARHGAP 31蛋白质表达未因突变而改变。功能分析表明，只有在两个突变体的共转染组细胞活力下降，细胞增殖受损，细胞凋亡被激活。Cdc 42活性在ARHGAP 31突变和FBLN 1突变单独作用下以及二者联合作用下均下降。此外，MAPK/ERK通路仅被两个突变体共转染组激活，而两个野生型转染组激活。 结论：我们报告了一个携带两种罕见的肢体缺陷相关基因ARHGAP 31和FBLN 1变体的病例，并提供了体外证据表明，这两种突变体引起的细胞功能的协同破坏可能会增强临床表现的特异性，扩大了我们对TTLD和其他遗传性疾病中因果基因相互作用的临床复杂性的认识。
Background: Aplasia cutis congenita (ACC) and terminal transverse limb defects (TTLDs) are the most common features of Adams-Oliver syndrome (AOS). ARHGAP31 is one of the causative genes for autosomal dominant forms of AOS, meanwhile its variants may only cause isolated TTLD. Here, we report a proband presented with apparent TTLD but not ACC. Methods: Whole exome sequencing (WES) and Sanger sequencing were applied to identify causative genes. Expression vectors were constructed for transfections in mammalian cell cultures followed by biochemical and functional analysis including immunoblotting, immunofluorescence staining, and cell counting kit-8 assay. Results: WES and Sanger sequencing suggested that the proband inherited rare ARHGAP31 variant [c.2623G > A (p.Glu875Lys)] and a rare FBLN1 variant [c.1649G > A (p.Arg550His)] from one of her asymptomatic parents, respectively. Given FBLN1 variation has also been linked to syndactyly, we suspected that the two genes together contributed to the TTLD phenotype and explored their possible roles in vitro. Mutant FBLN1 showed reduced expression resulted from impaired protein stability, whereas ARHGAP31 protein expression was unaltered by mutation. Functional assays showed that only in the co-transfected group of two mutants cell viability was decreased, cell proliferation was impaired, and apoptosis was activated. Cdc42 activity was declined by both ARHGAP31 mutation and FBLN1 mutation alone, and the two together. Furthermore, the MAPK/ERK pathway was only activated by two mutants co-transfected group compared with two wild-type transfections. Conclusion: We report a case carrying two rare variants of limb defects associated genes, ARHGAP31 and FBLN1, and provide in vitro evidence that synergistic disruption of cellular functions attributed by the two mutants may potentiate the penetrance of clinical manifestations, expanding our knowledge of clinical complexity of causal gene interactions in TTLD and other genetic disorders.
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影响因子：
5.2
作者：
Bohlega, Saeed;Al-Ajlan, Huda;Al-Saif, Amr
通讯作者：
Al-Saif, Amr
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影响因子：
5.5
作者：
Gong, Jiao;Jie, Yusheng;Chong, Yutian
通讯作者：
Chong, Yutian
影响因子：
2
作者：
Isrie, Mala;Wuyts, Wim;Devriendt, Koenraad
通讯作者：
Devriendt, Koenraad
影响因子：
3.9
作者：
Meester JAN;Sukalo M;Schröder KC;Schanze D;Baynam G;Borck G;Bramswig NC;Duman D;Gilbert-Dussardier B;Holder-Espinasse M;Itin P;Johnson DS;Joss S;Koillinen H;McKenzie F;Morton J;Nelle H;Reardon W;Roll C;Salih MA;Savarirayan R;Scurr I;Splitt M;Thompson E;Titheradge H;Travers CP;Van Maldergem L;Whiteford M;Wieczorek D;Vandeweyer G;Trembath R;Van Laer L;Loeys BL;Zenker M;Southgate L;Wuyts W
通讯作者：
Wuyts W
影响因子：
9.8
作者：
Southgate, Laura;Machado, Rajiv D.;Trembath, Richard C.
通讯作者：
Trembath, Richard C.