TY - JOUR
T1 - Targeting serous epithelial ovarian cancer with designer zinc finger transcription factors
AU - Lara, Haydee
AU - Wang, Yuhua
AU - Beltran, Adriana S.
AU - Juárez-Moreno, Karla
AU - Yuan, Xinni
AU - Kato, Sumie
AU - Leisewitz, Andrea V.
AU - Fredes, Mauricio Cuello
AU - Licea, Alexei F.
AU - Connolly, Denise C.
AU - Huang, Leaf
AU - Blancafort, Pilar
PY - 2012/8/24
Y1 - 2012/8/24
N2 - Ovarian cancer is the leading cause of death among gynecological malignancies. It is detected at late stages when the disease is spread through the abdominal cavity in a condition known as peritoneal carcinomatosis. Thus, there is an urgent need to develop novel therapeutic interventions to target advanced stages of ovarian cancer. Mammary serine protease inhibitor (Maspin) represents an important metastasis suppressor initially identified in breast cancer. Herein we have generated a sequence-specific zinc finger artificial transcription factor (ATF) to up-regulate the Maspin promoter in aggressive ovarian cancer cell lines and to interrogate the therapeutic potential of Maspin in ovarian cancer. We found that although Maspin was expressed in some primary ovarian tumors, the promoter was epigenetically silenced in cell lines derived from ascites. Transduction of the ATF in MOVCAR 5009 cells derived from ascitic cultures of a TgMISIIR-TAg mouse model of ovarian cancer resulted in tumor cell growth inhibition, impaired cell invasion, and severe disruption of actin cytoskeleton. Systemic delivery of lipid-protamine-RNA nanoparticles encapsulating a chemically modified ATF mRNA resulted in inhibition of ovarian cancer cell growth in nude mice accompanied with Maspin re-expression in the treated tumors. Gene expression microarrays of ATF-transduced cells revealed an exceptional specificity for the Maspin promoter. These analyses identified novel targets coregulated with Maspin in human short-term cultures derived from ascites, such as TSPAN12, that could mediate the anti-metastatic phenotype of the ATF. Our work outlined the first targeted, non-viral delivery of ATFs into tumors with potential clinical applications for metastatic ovarian cancers.
AB - Ovarian cancer is the leading cause of death among gynecological malignancies. It is detected at late stages when the disease is spread through the abdominal cavity in a condition known as peritoneal carcinomatosis. Thus, there is an urgent need to develop novel therapeutic interventions to target advanced stages of ovarian cancer. Mammary serine protease inhibitor (Maspin) represents an important metastasis suppressor initially identified in breast cancer. Herein we have generated a sequence-specific zinc finger artificial transcription factor (ATF) to up-regulate the Maspin promoter in aggressive ovarian cancer cell lines and to interrogate the therapeutic potential of Maspin in ovarian cancer. We found that although Maspin was expressed in some primary ovarian tumors, the promoter was epigenetically silenced in cell lines derived from ascites. Transduction of the ATF in MOVCAR 5009 cells derived from ascitic cultures of a TgMISIIR-TAg mouse model of ovarian cancer resulted in tumor cell growth inhibition, impaired cell invasion, and severe disruption of actin cytoskeleton. Systemic delivery of lipid-protamine-RNA nanoparticles encapsulating a chemically modified ATF mRNA resulted in inhibition of ovarian cancer cell growth in nude mice accompanied with Maspin re-expression in the treated tumors. Gene expression microarrays of ATF-transduced cells revealed an exceptional specificity for the Maspin promoter. These analyses identified novel targets coregulated with Maspin in human short-term cultures derived from ascites, such as TSPAN12, that could mediate the anti-metastatic phenotype of the ATF. Our work outlined the first targeted, non-viral delivery of ATFs into tumors with potential clinical applications for metastatic ovarian cancers.
KW - Actin Cytoskeleton/genetics
KW - Animals
KW - Cell Line, Tumor
KW - Female
KW - Gene Expression Regulation, Neoplastic
KW - Mice
KW - Mice, Transgenic
KW - Neoplasm Invasiveness
KW - Neoplasms, Experimental/genetics
KW - Oligonucleotide Array Sequence Analysis
KW - Ovarian Neoplasms/genetics
KW - RNA, Messenger/genetics
KW - Serpins/biosynthesis
KW - Tetraspanins/genetics
KW - Transcription Factors/biosynthesis
KW - Transcriptome
KW - Zinc Fingers
UR - http://www.scopus.com/inward/record.url?scp=84865469563&partnerID=8YFLogxK
U2 - 10.1074/jbc.M112.360768
DO - 10.1074/jbc.M112.360768
M3 - Article
C2 - 22782891
AN - SCOPUS:84865469563
SN - 0021-9258
VL - 287
SP - 29873
EP - 29886
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
IS - 35
ER -