Molecular and Functional Characterization of Three Different Postzygotic Mutations in PIK3CA-Related Overgrowth Spectrum (PROS) Patients: Effects on PI3K/AKT/mTOR Signaling and Sensitivity to PIK3 Inhibitors.
Molecular and Functional Characterization of Three Different Postzygotic Mutations in PIK3CA-Related Overgrowth Spectrum (PROS) Patients: Effects on PI3K/AKT/mTOR Signaling and Sensitivity to PIK3 Inhibitors.
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DOI：
10.1371/journal.pone.0123092
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Resta N
中科院分区：
文献类型：
--
作者：
Loconte DC;Grossi V;Bozzao C;Forte G;Bagnulo R;Stella A;Lastella P;Cutrone M;Benedicenti F;Susca FC;Patruno M;Varvara D;Germani A;Chessa L;Laforgia N;Tenconi R;Simone C;Resta N
PIK3CA相关的过度生长谱(PRO)包括一组仅影响肢体末端的疾病，例如I型巨指症，以及纤维脂肪过度生长(FAO)、巨脑-毛细血管畸形(MCAP)综合征、先天性脂肪瘤性躯干非对称性过度生长、淋巴管、毛细血管、静脉和混合型血管畸形、表皮痣、骨骼和脊柱异常(丁香)综合征和半增生性多发性脂肪瘤病(HHML)。杂合子后PIK3CA突变在这些综合征中经常被发现，而突变事件的时机和组织特异性可能是观察到的极端表型变异的原因。我们对3名患者(1名MCAP和2名FAO)进行了Sanger测序和PI3K/AKT/mTOR通路相关基因的定向深度测序，以确定导致突变的原因，并进行了免疫印迹分析，以检测受影响的皮肤成纤维细胞中AKT和p70S6K的磷酸化状态。此外，我们评估了它们在无血清情况下的生长能力以及它们对PI3K抑制剂Wortmannin和LY294002的体外反应。我们的数据表明，患者的细胞显示出PI3K/Akt通路的结构性激活。值得注意的是，PI3K的药物阻断导致培养中的增殖率显著降低，这表明抑制PI3K可能在未来对PROS患者的治疗中被证明是有益的。
PIK3CA-related overgrowth spectrum (PROS) include a group of disorders that affect only the terminal portion of a limb, such as type I macrodactyly, and conditions like fibroadipose overgrowth (FAO), megalencephaly-capillary malformation (MCAP) syndrome, congenital lipomatous asymmetric overgrowth of the trunk, lymphatic, capillary, venous, and combined-type vascular malformations, epidermal nevi, skeletal and spinal anomalies (CLOVES) syndrome and Hemihyperplasia Multiple Lipomatosis (HHML). Heterozygous postzygotic PIK3CA mutations are frequently identified in these syndromes, while timing and tissue specificity of the mutational event are likely responsible for the extreme phenotypic variability observed. We carried out a combination of Sanger sequencing and targeted deep sequencing of genes involved in the PI3K/AKT/mTOR pathway in three patients (1 MCAP and 2 FAO) to identify causative mutations, and performed immunoblot analyses to assay the phosphorylation status of AKT and P70S6K in affected dermal fibroblasts. In addition, we evaluated their ability to grow in the absence of serum and their response to the PI3K inhibitors wortmannin and LY294002 in vitro. Our data indicate that patients’ cells showed constitutive activation of the PI3K/Akt pathway. Of note, PI3K pharmacological blockade resulted in a significant reduction of the proliferation rate in culture, suggesting that inhibition of PI3K might prove beneficial in future therapies for PROS patients.
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影响因子：
30.8
作者：
Lindhurst, Marjorie J.;Parker, Victoria E. R.;Payne, Felicity;Sapp, Julie C.;Rudge, Simon;Harris, Julie;Witkowski, Alison M.;Zhang, Qifeng;Groeneveld, Matthijs P.;Scott, Carol E.;Daly, Allan;Huson, Susan M.;Tosi, Laura L.;Cunningham, Michael L.;Darling, Thomas N.;Geer, Joseph;Gucev, Zoran;Sutton, V. Reid;Tziotzios, Christos;Dixon, Adrian K.;Helliwell, Timothy;O'Rahilly, Stephen;Savage, David B.;Wakelam, Michael J. O.;Barroso, Ines;Biesecker, Leslie G.;Semple, Robert K.
通讯作者：
Semple, Robert K.
影响因子：
2
作者：
Keppler-Noreuil, Kim M.;Sapp, Julie C.;Lindhurst, Marjorie J.;Parker, Victoria E. R.;Blumhorst, Cathy;Darling, Thomas;Tosi, Laura L.;Huson, Susan M.;Whitehouse, Richard W.;Jakkula, Eveliina;Grant, Ian;Balasubramanian, Meena;Chandler, Kate E.;Fraser, Jamie L.;Gucev, Zoran;Crow, Yanick J.;Brennan, Leslie Manace;Clark, Robin;Sellars, Elizabeth A.;Pena, Loren D. M.;Krishnamurty, Vidya;Shuen, Andrew;Braverman, Nancy;Cunningham, Michael L.;Sutton, V. Reid;Tasic, Velibor;Graham, John M., Jr.;Geer, Joseph, Jr.;Henderson, Alex;Semple, Robert K.;Biesecker, Leslie G.
通讯作者：
Biesecker, Leslie G.
影响因子：
7.3
作者：
Pezze, Piero Dalle;Sonntag, Annika G.;Thedieck, Kathrin
通讯作者：
Thedieck, Kathrin
影响因子：
9.8
作者：
Kurek, Kyle C.;Luks, Valerie L.;Warman, Matthew L.
通讯作者：
Warman, Matthew L.
影响因子：
4.8
作者：
Gan, Xiaoqing;Wang, Jiyong;Wu, Dianqing
通讯作者：
Wu, Dianqing