Synergistic effect of PKC activation and actin filament disruption on carbonate apatite-facilitated lymphocyte transfection

• Published in: Biochemical and biophysical research communications Vol. 419; no. 3; pp. 482 - 484
• Main Authors: Fukuda, K., Kutsuzawa, K., Maruyama, K., Akiyama, Y., Chowdhury, E.H.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 16.03.2012
• Subjects: Actin Cytoskeleton - metabolism; Actin disruption; Apatites; Ca2+ ionophore; Carbonate apatite; DNA uptake; Endocytosis; Enzyme Activation; Genetic Therapy - methods; HeLa Cells; Humans; Jurkat Cells; Leukemia - therapy; Lymphocytes - drug effects; Lymphocytes - metabolism; Nano-crystals; Nanoparticles; PKC activation; Protein Kinase C - biosynthesis; T-leukemia cell transfection; Transfection - methods; DMEM; Endocytosis; PMA; Carbonate apatite; Ca2+ ionophore; PKC; Nano-crystals; RLU; T-leukemia cell transfection; DNA uptake; Actin disruption; PKC activation
• ISSN: 0006-291X; 1090-2104
• DOI: 10.1016/j.bbrc.2012.02.023

► PKC activation enhances carbonate apatite-facilitated lymphocyte transfection. ► Actin filament disruptor synergizes with PKC activators for accelerating transfection efficacy. ► PKC activation might promote particle endocytosis through specific integrins and syndecan. 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 Ca2+ 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.