SUMOylation regulates LKB1 localization and its oncogenic activity in liver cancer

Imanol Zubiete-Franco (1) , Juan García-Rodríguez (1) , Fernando Lopitz-Otsoa (1) , Marina Serrano-Macia (1) , Jorge Simon (1) , Pablo Fernández-Tussy (1) , Lucía Barbier-Torres (1) , David Fernández-Ramos (1) , Virginia Gutiérrez-De-Juan (1) , Sergio López de Davalillo (1) , Onintza Carlevaris , Adolfo Beguiristain Gómez , Erica Villa , Diego Calvisi , César Martín , Edurne Berra (2) , Patricia Aspichueta , Naiara Beraza (1) , Marta Varela-Rey (1) , Matias Ávila , Manuel S. Rodríguez (3, 4) , José Mato (1) , Irene Díaz-Moreno (5) , Antonio Díaz-Quintana (5) , Teresa Delgado (1) , María Martínez-Chantar (1)

1 CIBERehd - Centro de Investigación Biomédica en Red en el Área temática de Enfermedades Hepáticas y Digestivas
2 IBDC - Institut de signalisation, biologie du développement et cancer - Institute of Developmental Biology and Cancer
3 IPBS - Institut de pharmacologie et de biologie structurale
4 ITAV - Institut des Technologies Avancées en sciences du Vivant
5 Instituto de Investigaciones Químicas (IIQ)

Background: Even though liver kinase B1 (LKB1) is usually described as a tumor suppressor in a wide variety of tissues, it has been shown that LKB1 aberrant expression is associated with bad prognosis in Hepatocellular Carcinoma (HCC). Methods: Herein we have overexpressed LKB1 in human hepatoma cells and by using histidine pull-down assay we have investigated the role of the hypoxia-related post-translational modification of Small Ubiquitin-related Modifier (SUMO)ylation in the regulation of LKB1 oncogenic role. Molecular modelling between LKB1 and its interactors, involved in regulation of LKB1 nucleocytoplasmic shuttling and LKB1 activity, was performed. Finally, high affinity SUMO binding entities-based technology were used to validate our findings in a pre-clinical mouse model and in clinical HCC. Findings: We found that in human hepatoma cells under hypoxic stress, LKB1 overexpression increases cell viability and aggressiveness in association with changes in LKB1 cellular localization. Moreover, by using site-directed mutagenesis, we have shown that LKB1 is SUMOylated by SUMO-2 at Lys178 hampering LKB1 nucleocytoplasmic shuttling and fueling hepatoma cell growth. Molecular modelling of SUMO modified LKB1 further confirmed steric impedance between SUMOylated LKB1 and the STe20-Related ADaptor cofactor (STRADα), involved in LKB1 export from the nucleus. Finally, we provide evidence that endogenous LKB1 is modified by SUMO in pre-clinical mouse models of HCC and clinical HCC, where LKB1 SUMOylation is higher in fast growing tumors. Interpretation: Overall, SUMO-2 modification of LKB1 at Lys178 mediates LKB1 cellular localization and its onco-genic role in liver cancer. und: This work was supported by grants from NIH (US Department of Health and Human services)-R01AR001576-11A1 (J.M.M and M.L.M-C.), Gobierno Vasco-Departamento de Salud 2013111114 (to M.L.M.-C), ELKARTEK 2016, Departamento de Industria del Gobierno Vasco (to M.L.M.-C), MINECO: SAF2017-87301-R and SAF2014-52097-R integrado en el Plan Estatal de Investigación Cientifica y Técnica y Innovación 2013-2016 cofinanciado con Fondos FEDER (to M.L.M.-C and J.M.M., respectively), BFU2015-71017/BMC Keywords: LKB1 SUMO HCC SIRT1 STRADα EBioMedicine 40 (2019) 406-421 ⁎ Corresponding authors at: CIC bioGUNE, Ed. 801A Parque Tecnológico de Bizkaia,

Mots clés: LKB1; SUMO; HCC; SIRT1; STRADα

Domaines: Cancer; Hépatologie et Gastroentérologie; Génomique, Transcriptomique et Protéomique [q-bio.GN]

Imanol Zubiete-Franco, Juan García-Rodríguez, Fernando Lopitz-Otsoa, Marina Serrano-Macia, Jorge Simon, et al.. SUMOylation regulates LKB1 localization and its oncogenic activity in liver cancer. EBioMedicine, 2019, 40, pp.406-421. ⟨10.1016/j.ebiom.2018.12.031⟩. ⟨hal-02351178⟩