Synaptojanin 2 and microRNA-31 control invadopodia formation and metastasis by regulating vesicular trafficking

Nir Ben-Chetrit 1 David Chetrit 3 Cindy Körner 4 Maicol Mancini 1 Tomer Itkin 2 Silvia Carvalho 1 Hadas Cohen-Dvashi 1 Wolfgang Koestller 1 Kirti Sharma 4 Moshit Lindzen 1 Ziv Shulman 2 Raanan Margalit 2 Dalia Seger 1 Hannah Schmidt-Glenewinkel 1 Daniela Ferraro 1 Fresia Pareja 1 Martine Bernstein 1 Hava Gil-Henn 5 Tsvee Lapidot 2 Ronen Alon 2 Fernanda Milanezi 6 Marc Symons 7 Fernando Schmitt 6 Stefan Wiemann 4 Marcelo Ehrlich 3 Yosef Yarden 1
1Department of Biological Regulation, Weizmann Institute of Science, Rehovot
2Department of Immunology, Weizmann Institute of Science, Rehovot
3Department of Cell Research and Immunology, Tel-Aviv University, Tel-Aviv
4Division of Molecular Genome Analysis, German Cancer Research Centre (DKFZ), Heidelberg
5Faculty of Medicine, Bar-Ilan University, Safed
6IPATIMUP, University of Porto, Porto
7Center for Oncology and Cell Biology, The Feinstein Institute for Medical Research, New York
Growth factors propel cell migration in vitro and metastasis in vivo, but the underlying mechanisms are incompletely understood. Employing EGF-stimulated mammary cells we linked the lipid phosphatase synaptojanin 2 (SYNJ2) to MAPK-dependent invasive phenotype, as well as demonstrate high SYNJ2 expression in aggressive human breast tumors and low survival rates. High expression of SYNJ2 in benign or tumorigenic mammary epithelial cells, results with stabilization of EGFR and sustained MAPK signaling.In addition, high expression in mammary and other tumors might relate to repression of microRNA-31, a metastasis suppressor able to restrain SYNJ2 expression.
Knockdown of SYNJ2 in mammary tumor cells attenuates MAPK signaling and almost abolished their intravasation into blood vessels, metastasis to lymph nodes and lung colonization. When tested in vitro, SYNJ2-depleted cells exhibited deformed focal adhesions and disappearance of invadopodia. These effects correlated with derailed trafficking of both EGFR and beta-1 integrin, as well as defective delivery of metalloproteinases to invadopodia. We conclude that recycling of active EGFRs promotes invadopodia formation by locally dephosphorylating phosphatidyl-inositol 3,4,5-trisphosphate, PI(3,4,5)P3, into PI(3,4)P2, thereby priming invadopodia formation. Because of their emerging roles in metastasis, dephosphorylation of phosphoinositides and vesicular trafficking might serve as targets for cancer therapy.








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