Establishment of zebrafish model via TIE2-R849W and exploration of its role in hereditary venous malformations

doi:10.19438/j.cjoms.2018.01.002

Abstract

Abstract: PURPOSE: To explore the role of TIE2-R849W, a mutation related to hereditary venous malformations, via establishing a novel zebrafish model. METHODS: Through the designment and synthesis of TIE2-R849W expression plasmid, translated mRNA was microinjected into fli1a: EGFP fertilized eggs, which were used to establish a TIE2-R849W over-expression model. Under fluorescence microscope, the developmental change of caudal vein plexus (CVP) was recorded and quantitatively compared. Using RNA extracted from zebrafish, target genes were tested by real-time quantitative PCR (qRT-PCR). The existing gene chip was analyzed to further verify the results. Statistical analysis was performed with GraphPad Prism 5. RESULTS: Based on observed phenotypes of TIE2-R849W over-expression zebrafish, TIE2-R849W significantly affected the development of the caudal venous plexus, which displayed a significant decrease in the number and area of sinus. According to the results of qRT-PCR, upregulation of pik3r2 and foxo1b could be induced by TIE2-R894W over-expression. At the same time, except for significantly up-regulated egfl7, expression of nr2f2, nr2f1a and s1pr1 had no significant difference from the control. In contrast to in vitro chip analysis, tie2, pik3r2, foxo1b and egfl7 showed consistent high expression in individual mutant group. CONCLUSIONS: TIE2-R849W associated with hereditary venous malformations could induce abnormal development of zebrafish CVP, which implies a possible relationship between VM and egfl7.

Key words: Venous malformations, Caudal vein plexus, Zebrafish, Egfl7

CLC Number:

R543.6

DU Zhong, MA Hai-long, ZHANG Ling, ZHENG Jia-wei, WANG Yan-an. Establishment of zebrafish model via TIE2-R849W and exploration of its role in hereditary venous malformations[J]. China Journal of Oral and Maxillofacial Surgery, 2018, 16(1): 6-11.

References

[1] Boscolo E, Limaye N, Huang L, et al.Rapamycin improves TIE2-mutated venous malformation in murine model and human subjects[J]. J Clin Invest, 2015, 125(9): 3491-3504.
[2] 郑家伟, 张凌, 陈正岗. 脉管异常的ISSVA新分类[J]. 中国口腔颌面外科杂志, 2015, 13(5): 385-388.
[3] Shu W, Lin Y, Hua R, et al.Cutaneomucosal venous malformations are linked to the TIE2 mutation in a large Chinese family[J]. Exp Dermatol, 2012, 21(6): 456-457.
[4] 舒伟, 林有坤, 华荣, 等. 一个中国汉族皮肤和黏膜多发静脉血管畸形家系的单倍型分析[J]. 遗传, 2012, 34(4): 431-436.
[5] Calvert JT, Riney TJ, Kontos CD, et al.Allelic and locus heterogeneity in inherited venous malformations[J]. Hum Mol Genet, 1999, 8(7): 1279-1289.
[6] Nätynki M, Kangas J, Miinalainen I, et al.Common and specific effects of TIE2 mutations causing venous malformations[J]. Hum Mol Genet, 2015, 24(22): 6374-6389.
[7] Uebelhoer M, Nätynki M, Kangas J, et al.Venous malformation-causative TIE2 mutations mediate an AKT-dependent decrease in PDGFB[J]. Hum Mol Genet, 2013, 22(17): 3438-3448.
[8] 李礼, 罗凌飞. 以斑马鱼为模式动物研究器官的发育与再生[J]. 遗传, 2013, 35(4): 321-332.
[9] Wakayama Y, Fukuhara S, Ando K, et al.Cdc42 mediates Bmp-induced sprouting angiogenesis through Fmnl3-driven assembly of endothelial filopodia in zebrafish[J]. Dev Cell, 2015, 32(1): 109-122.
[10] Deng W, Wang Y, Liu Z, et al.HemI: a toolkit for illustrating heatmaps[J]. PLoS One, 2014, 9(11): e111988.
[11] Huang YH, Wu MP, Pan SC, et al.STAT1 activation by venous malformations mutant Tie2-R849W antagonizes VEGF-A-mediated angiogenic response partly via reduced bFGF production[J]. Angiogenesis, 2013, 16(1): 207-222.
[12] Kashiwada T, Fukuhara S, Terai K, et al.β-Catenin-dependent transcription is central to Bmp-mediated formation of venous vessels[J]. Development, 2015, 142(3): 497-509.
[13] Wu BJ, Chiu CC, Chen CL, et al.Nuclear receptor subfamily 2 group F member 1a (nr2f1a) is required for vascular development in zebrafish[J]. PLoS One, 2014, 9(8): e105939.
[14] Mendelson K, Zygmunt T, Torres-Vázquez J, et al.Sphingosine 1-phosphate receptor signaling regulates proper embryonic vascular patterning[J]. J Biol Chem, 2013, 288(4): 2143-2156.
[15] Nikolic I, Stankovic ND, Bicker F, et al.EGFL7 ligates αvβ3 integrin to enhance vessel formation[J]. Blood, 2013, 121(15): 3041-3050.
[16] Poissonnier L, Villain G, Soncin F, et al.Egfl7 is differentially expressed in arteries and veins during retinal vascular development[J]. PLoS One, 2014, 9(3): e90455.
[17] Deford P, Brown K, Richards RL, et al.MAGP2 controls Notch via interactions with RGD binding integrins: Identification of a novel ECM-integrin-Notch signaling axis[J]. Exp Cell Res, 2016, 341(1): 84-91.
[18] Castel P, Carmona FJ, Grego-Bessa J, et al. Somatic PIK3CA mutations as a driver of sporadic venous malformations [J]. Sci Transl Med, 2016, 8(332): 332ra42.
[19] Castillo SD, Tzouanacou E, Zaw-Thin M, et al. Somatic activating mutations in Pik3ca cause sporadic venous malformations in mice and humans [J]. Sci Transl Med, 2016, 8(332): 332ra43.
[20] Renz M, Otten C, Faurobert E, et al.Regulation of β1 integrin-Klf2-mediated angiogenesis by CCM proteins[J]. Dev Cell, 2015, 32(2): 181-190.
[21] Skon CN, Lee JY, Anderson KG, et al.Transcriptional downregulation of S1pr1 is required for the establishment of resident memory CD8+ T cells[J]. Nat Immunol, 2013, 14(12): 1285-1293.