Factors produced by hepatocellular carcinoma micronevironment induce changes in human mesenchymal stromal cells.

GARCÍA, MARIANA G.; BAYO, JUAN; REAL, ALEJANDRINA; FIORE, ESTEBAN; MALVICINI, MARIANA ; PICCIONI, FLAVIA; BOLONTRADE, MARCELA F.; ANDRIANI, OSCAR; BIZAMA, CAROLINA ; FRESNO, CRISTÓBAL; PODHAJCER, OSVALDO. ; FERNÁNDEZ, ELMER; GIDEKEL, MANUEL.; MAZZOLINI, GUILLERMO

Estocolmo

Simposio; 13th Annual Meeting- International Society for Stem Cell Research (ISSCR); 2015

International Society for Stem Cell Research (ISSCR)

Hepatocellular carcinoma (HCC) is the 2nd cause of cancer-related death and the majority of patients are diagnosed at advanced stages. New therapies are needed and those focused on the delivery therapeutic genes by mesenchymal stromal cells (MSC)are gaining interest. HCC produces factors that recruit MSC to the tumor. However, after MSC homing, these factors could modify MSC biology. The aim of this work was to study changes occurred in human MSC exposed to conditioned media (CM)derived from HCC tumors. Cytokines in CM derived from HCC tumors were examined by an antibody array. CM were obtained from fresh human HCC samples (PT-7, PT-12 and PT-19) or tumors subcutaneously generated by a primary culture from a patient with HCC (HC-PT-5) or HuH7 cell line in nude mice. All CM demonstrated similar expression of cytokines including GRO,MCP-1 and IL-8 being the latter with the highest concentration.We evaluated their role in MSC chemotaxis to HCC by a Boydenchamber using neutralizing antibodies anti-CXCR1, CXCR2 orMCP-1. Anti-MCP-1 reduced MSC migration towards CM from HCC around 20%. Similarly, anti-CXCR1 or CXCR2 reduced MSC in vitro migration to HCC approximately 20%. When both receptors were neutralized together MSC migration was reduced about 50%. To evaluate changes on MSC behavior by HCC released factors, MSC were pre-stimulated for 24 h with CM from HuH7 or HCPT-5. Chemotaxis assay showed a 2-fold increase of stimulated MSC(sMSC) migration towards the CM in comparison with unstimulated MSC (usMSC). Gene expression profile of sMSC with CM from PT-7,PT-12, HC-PT-5 or HuH7 tumors showed that MSC differentially expressed 445, 511, 521 and 511 genes respectively in comparison with usMSC. Among these genes 46 are related with cell migration and invasion. The most important chemokines produced by sMSC were MCP-1, Nap2, IP-10, Mip1β and Eotaxin-2. Factors produced by sMSC were able to increase fibroblasts and endothelial cellschemotaxis in comparison to factors produced by usMSC. Finally,sMSC with HuH7 CM and then inoculated in HCC tumor bearing mice did not modify tumor growth. We can conclude that factors produced by HCC are Responsible from MSC chemotaxis and induce changes in MSC gene profile, in the released factors and in their chemotactic capacity. However, these changes did not affect the aggressiveness of HCC in vivo.