title: STAT3/NF-κB-Regulated Lentiviral TK/GCV Suicide Gene Therapy for Cisplatin-Resistant Triple-Negative Breast Cancer.
abstract: Triple-negative breast cancer (TNBC) represents approximately 20% of all breast cancers and appears resistance to conventional cytotoxic chemotherapy, demonstrating a particularly poor prognosis and a significantly worse clinical outcome than other types of cancer. Suicide gene therapy has been used for the in vivo treatment of various solid tumors in recent clinical trials. In tumor microenvironment, STAT3/NF-κB pathways are constitutively activated in stromal cells as well as in cancer stem cells (CSCs). In this study, we have cloned a novel STAT3/NF-κB-based reporter system to drive the expression of herpes simplex virus thymidine kinase (HSV-TK) against breast cancer. Lentiviral vector expressing HSV-TK under the regulation of STAT3/NF-κB fused response element was developed. In this setting, we exploited the constitutive STAT3/NF-κB activation in tumors to achieve higher transgene expression than that driven by a constitutively active CMV promotorin vivo. An orthotropic MDA-MB-231 triple negative breast cancer mouse model was used for evaluating the feasibility of STAT3-NF-κB-TK/GCV suicide gene therapy system. The basal promoter activity of Lenti-CMV-TK and Lenti-STAT3-NF-κB-TK in MDA-MB-231 cells was compared by3H-FEAU uptake assay. The Lenti-CMV-TK showed ~5 fold higher3H-FEAU uptake then Lenti -STAT3-NF-κB-TK. In clonogenic assay, cells expressing Lenti-CMV-TK were 2-fold more sensitive to GCV than Lenti-STAT3-NF-κB-TK transduced cells.In vitroeffect of STAT3-NF-κB-induced transgene expression was determined by 10ng/mL TNF-α induction and confirmed by western blot analysis and DsRedm fluorescent microscopy.In vivoevaluation of therapeutic effect by bioluminescence and [18F]FHBG microPET imaging indicated that Lenti-STAT3-NF-κB-TK showed more tumor growth retardation than Lenti-CMV-TK when GCV (20 mg/kg) was administered. The invasiveness and expression of cancer stem cell markers were both decreased after STAT3/NF-κB-regulated HSV-TK/GCV therapy. Moreover, STAT3/NF-κB signaling targeting could further sensitize tumor cells to cisplatin. This study successfully established a theranositic approach to treat triple-negative breast cancer via STAT3-NF-κB responsive element-driven suicide gene therapy. This platform may also be an alternative strategy to handle with drug-resistant cancer cells.
authors.affiliation: Department of Biomedical Imaging and Radiological Sciences, School of Biomedical and Engineering, National Yang-Ming University, Taipei, Taiwan.
authors.forename: Wei-Ying
authors.initials: WY
authors.lastname: Kuo
authors.affiliation: Molecular and Genetic Imaging Core/Taiwan Mouse Clinic, NRPB, Department of Nuclear Medicine and National PET/Cyclotron Center, Taipei Veterans General Hospital, Taipei, Taiwan.
authors.forename: Luen
authors.initials: L
authors.lastname: Hwu
authors.affiliation: Department of Biomedical Imaging and Radiological Science, China Medical University, Taichung, Taiwan.
authors.forename: Chun-Yi
authors.initials: CY
authors.lastname: Wu
authors.affiliation: Department of Biomedical Imaging and Radiological Sciences, School of Biomedical and Engineering, National Yang-Ming University, Taipei, Taiwan.
authors.forename: Jhih-Shian
authors.initials: JS
authors.lastname: Lee
authors.affiliation: Department of Nuclear Medicine, Taipei Veterans General Hospital, Taipei, Taiwan.
authors.forename: Chi-Wei
authors.initials: CW
authors.lastname: Chang
authors.affiliation: Department of Nuclear Medicine, Taipei Veterans General Hospital, Taipei, Taiwan.; Biomedical Imaging Research Center, Department of Biomedical Imaging and Radiological Sciences, School of Biomedical and Engineering, Institute of Clinical Medicine, School of Medicine, National Yang-Ming University.
authors.forename: Ren-Shyan
authors.initials: RS
authors.lastname: Liu
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000008:administration & dosage*; D002945:Cisplatin / Q000008:administration & dosage; D004195:Disease Models, Animal; D004351:Drug Resistance; D015774:Ganciclovir / Q000008:administration & dosage*; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D016086:Lentivirus / Q000235:genetics; D051379:Mice; D052002:NF-kappa B p50 Subunit / Q000235:genetics / Q000378:metabolism*; D050796:STAT3 Transcription Factor / Q000235:genetics / Q000378:metabolism*; D013937:Thymidine Kinase / Q000235:genetics / Q000378:metabolism*; D014161:Transduction, Genetic; D016896:Treatment Outcome; D064726:Triple Negative Breast Neoplasms / Q000628:therapy*
chemical_list: D000970:Antineoplastic Agents; D052002:NF-kappa B p50 Subunit; D050796:STAT3 Transcription Factor; C494087:Stat3 protein, mouse; C073701:Nfkb1 protein, mouse; D013937:Thymidine Kinase; D015774:Ganciclovir; D002945:Cisplatin
keywords: Cisplatin resistance.; TK/GCV suicide gene therapy; Triple-negative breast cancer