Files

No attached file found for this publication.

Details

Authors
Abstract
Objectives: Germline substitutions in the endothelial cell tyrosine kinase receptor TIE2/TEK cause a rare inherited form of venous anomalies, Mucocutanous Venous Malformation (VMCM). Moreover, at least 50% of common sporadic venous malformations (VM) are caused by somatic mutations in TIE2. The identification of the etiopathogenic cause forms the basis for the generation of in vivo mouse models of the disease. Methods: Conditional “knock-in” lines carrying the most frequent somatic Tie2 mutation, L914F, have been generated. The targeting construct replaces wild-type exon 17 with cDNA starting at exon 17, followed by the remaining 3’ coding region flanked by loxP sites, and then the mutant exon. Upon Cre-expression, the cDNA containing the wild-type exon is floxed out, leaving only the mutant exon and endogenous 3’ exons to be expressed. After electroporation, homologously recombined murine ES cells were identified using PCR and Southern blot, and selected clones were injected into blastocysts to generate targeted mice. Results: Eight chimeras have been obtained, and germline transmission of the targeted locus was confirmed. Ubiquitous Cre-expression, causing wild-type Tie2 excision, is being induced by crossing with PGK-Cre mice. Preliminary results indicate embryonic lethality before E10.5. Tamoxifen-inducible, endothelial cell-specific replacement will be obtained by crossing with Cdh5-CreERT2 transgenics. Conclusion: The knock-in strategy replaces wild-type Tie2 with VM-causative mutations, hopefully generating an in vivo model of the disease. This will allow us to study the effects of the mutations and better understand their etiopathogenic mechanisms, as well as test novel therapeutic measures.
Affiliations

Citations

  • APA
  • Chicago
  • FWB

Uebelhoer, M., Brouillard, P., Achouri, Y., & Vikkula, M. (2011). Towards a Venous Malformation Mouse Model. 11th Annual Meeting of the Belgian Society of Human Gentics, Louvain-la-Neuve, Belgium. https://hdl.handle.net/2078.5/55611