title: Synergistic effect of PKC activation and actin filament disruption on carbonate apatite-facilitated lymphocyte transfection.
abstract: Leukemia and lymphoma cells are potential targets for genetic manipulation in cancer therapy. On the other hand, genetically modified autologous lymphocytes expressing a chimeric antigen against a receptor overexpressed in tumor cells or tumor vasculature are promising cell-based therapeutics for cancer.However, the lack of a smart device for efficient transgene delivery to the lymphocytes poses the major obstacle to the successful clinical applications of these attractive approaches. Recently, we developed a carbonate apatite-based nanocarrier system for effective intracellular delivery and release of DNA molecules, achieving very high level of transgene expression in both primary and cancer cells. Although its efficacy in human T leukemia cells is relatively poor, immobilization of fibronectin and/or chimeric E-cadherin-Fc on particle surface could enhance transgene delivery in presence of an actin filament disrupter. Here, we report for the first time that simultaneous stimulation of human T leukemia cells by a protein kinase C (PKC) activator, a Ca(2+) ionophore and an actin filament disrupter dramatically accelerated carbonate apatite-mediated transgene delivery in the cells, resulting in over 100-fold more efficacy than commcercially available lipofectamine.
authors.affiliation: Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Fukuda
authors.forename: K
authors.initials: K
authors.lastname: Kutsuzawa
authors.forename: K
authors.initials: K
authors.lastname: Maruyama
authors.forename: Y
authors.initials: Y
authors.lastname: Akiyama
authors.forename: E H
authors.initials: EH
authors.lastname: Chowdhury
mesh_terms: D008841:Actin Cytoskeleton / Q000378:metabolism*; D001031:Apatites; D004789:Enzyme Activation; D015316:Genetic Therapy / Q000379:methods*; D006367:HeLa Cells; D006801:Humans; D019169:Jurkat Cells; D007938:Leukemia / Q000628:therapy; D008214:Lymphocytes / Q000187:drug effects / Q000378:metabolism*; D053758:Nanoparticles; D011493:Protein Kinase C / Q000096:biosynthesis*; D014162:Transfection / Q000379:methods*
chemical_list: D001031:Apatites; C030782:carboapatite; D011493:Protein Kinase C