ABSTRACT
The cell surface transmembrane receptor TM4SF5 has been implicated in hepatocellular carcinoma (HCC), but its candidacy as a therapeutic target has not been evaluated. Building on findings that immunization with a peptide vaccine targeting human TM4SF5 can exert prophylactic and therapeutic effects in a murine model of HCC, we developed a monoclonal antibody to characterize expression of TM4SF5 in HCC and to target its function there as an anticancer strategy. We found that the antibody modulated cell signaling in HCC cells in vitro, reducing cell motility, modulating E-cadherin expression, altering p27(kip1) localization, and increasing RhoA activity. Using a mouse xenograft model of human HCC, we documented the in vivo efficacy of the antibody, which suppressed tumor growth in either tumor prevention or treatment designs. Our work offers a preclinical proof of concept for TM4SF5 as a promising target for antibody therapeutics to treat HCC. Cancer Res; 74(14); 3844-56. ©2014 AACR.
Subject(s)
Antibodies, Monoclonal/pharmacology , Antineoplastic Agents/pharmacology , Carcinoma, Hepatocellular/metabolism , Carcinoma, Hepatocellular/pathology , Liver Neoplasms/metabolism , Liver Neoplasms/pathology , Membrane Proteins/antagonists & inhibitors , Actins/metabolism , Animals , Antibodies, Monoclonal/administration & dosage , Antibody Specificity , Antineoplastic Agents/administration & dosage , Cadherins/metabolism , Carcinoma, Hepatocellular/genetics , Cell Line, Tumor , Cell Movement/drug effects , Cell Nucleus/metabolism , Cyclin-Dependent Kinase Inhibitor p27/metabolism , Disease Models, Animal , Gene Expression , Humans , Liver Neoplasms/genetics , Male , Membrane Proteins/genetics , Membrane Proteins/metabolism , Mice , Organ Specificity , Protein Transport , Tumor Burden/drug effects , Xenograft Model Antitumor Assays , rho GTP-Binding Proteins/metabolismABSTRACT
The innovation of a peptide vaccine strategy may contribute to the development of efficacious and convenient cancer vaccines. Recently, we formulated an efficacious peptide vaccine without carriers using the natural phosphodiester bond CpG-DNA and a special liposome complex [Lipoplex(O)]. The peptide vaccine targeting a tumor antigen, transmembrane 4 superfamily member 5 protein (TM4SF5), was confirmed to have preventive and therapeutic effects in a mouse hepatocellular carcinoma (HCC) model. In this study, we demonstrated that the isotype-switched (IgM(-)IgD(-)) B cell population increased after immunization and that the functional memory response persisted for at least 70 days after the final immunization of mice. Delayed implantation of BNL-HCC cells significantly induced the peptide-specific IgG2a production in the immunized mice. Accordingly, tumor growth was inhibited and the survival rate increased. These results suggest that our peptide vaccine induces memory response, which is essential for cancer vaccine application.
Subject(s)
B-Lymphocytes/immunology , Cancer Vaccines/immunology , Carcinoma, Hepatocellular/immunology , Immunologic Memory/immunology , Liver Neoplasms/immunology , Membrane Proteins/immunology , Animals , Cancer Vaccines/therapeutic use , Carcinoma, Hepatocellular/prevention & control , Cell Line, Tumor , CpG Islands/immunology , Disease Models, Animal , Epitopes/immunology , Immunoglobulin Class Switching , Immunoglobulin G/immunology , Kaplan-Meier Estimate , Liposomes , Liver Neoplasms/prevention & control , Male , MiceABSTRACT
Human embryonic stem (ES) cells are a potential source of cells for developmental studies and for a variety of applications in transplantation therapies and drug discovery. However, human ES cells are difficult to culture and maintain at a large scale, which is one of the most serious obstacles in human ES cell research. Culture of human ES cells on MEF cells after disassociation with accutase has previously been demonstrated by other research groups. Here, we confirmed that human ES cells (H9) can maintain stem cell properties when the cells are passaged as single cells under a feeder-free culture condition. Accutase-dissociated human ES cells showed normal karyotype, stem cell marker expression, and morphology. We prepared frozen stocks during the culture period, thawed two of the human ES cell stocks, and analyzed the cells after culture with the same method. Although the cells revealed normal expression of stem cell marker genes, they had abnormal karyotypes. Therefore, we suggest that accutase-dissociated single cells can be usefully expanded in a feeder-free condition but chromosomal modification should be considered in the culture after freeze-thawing.
