RESUMEN
Assembly morphology made from lipids is controlled by the balance between the polar headgroup and the hydrophobic tails. In this study, we showed the various generations of polyamidoamine dendron-bearing lipids could form various assembly morphologies. Furthermore, the effect of the assembly morphologies made from dendron-bearing lipids for transfection abilities were examined. We synthesized three novel dendron-bearing lipids, DL-U2-G1 (G1), DL-U2-G2 (G2), and DL-U2-G3 (G3), which included first, second, and third generation polyamidoamine dendrons, respectively. Transmission electron microscopy showed that lipoplexes (complexes with cationic lipids and plasmid DNA) comprising G1 had multilamellar structures. G2 presented as aggregates of cubic particles and G3 exhibited clusters of spherical micelles. The ability to form complexes with plasmid DNA was in the decreasing order G3 > G2 > G1; calcein release from endosomes was in the order G3 > G2, G1; and transfection activity followed the order G1 > G2, G3. Interaction of the lipoplexes with heparin suggests that G3 had a lower level of plasmid DNA dissociation from lipoplexes than G1 in vitro. These results suggest that the size of the DL-U2 headgroup determines assembly morphology and that the structure markedly affects transfection activity.
Asunto(s)
ADN/administración & dosificación , Dendrímeros/química , Lípidos/química , Plásmidos/administración & dosificación , Transfección , Animales , Células COS , Chlorocebus aethiops , ADN/genética , Dendrímeros/síntesis química , Células HeLa , Humanos , Lípidos/síntesis química , Plásmidos/genéticaRESUMEN
Active targeting of the liposome is an attractive strategy for drug delivery and in vivo bio-imaging. We previously reported the specific accumulation of Sialyl Lewis X (SLX) liposome to inflamed tissue in arthritic model mice or tumor-bearing mice. SLX-liposome encapsulation with fluorescent substances allows for the visualization of these liposomes by the time-dependent transvascular accumulation of fluorescent signals in the histological sections. In the present study, we developed a new SLX-liposome encapsulated with colloidal gold for transmission electron microscopic observation. We herein describe the characterization of the colloidal gold-loaded SLX-liposomes and demonstrate its specific targeting to the endothelial cells of tumor blood vessels in tumor-bearing mice.
Asunto(s)
Células Endoteliales/ultraestructura , Oro Coloide/metabolismo , Liposomas/farmacocinética , Microscopía Electrónica de Transmisión/métodos , Neoplasias/irrigación sanguínea , Animales , Artritis/metabolismo , Sistemas de Liberación de Medicamentos/métodos , Femenino , Inflamación/metabolismo , Liposomas/ultraestructura , Ratones , Ratones Endogámicos BALB C , Modelos Animales , Neoplasias/ultraestructura , Oligosacáridos/metabolismo , Antígeno Sialil Lewis XRESUMEN
Generally angiogenic factors induce the expression of E-selectin in vascular endothelial cells in the tumors. In this study, we employed an anti-E-selectin monoclonal antibody to target tumors in vivo and evaluated an optical imaging reagent to visualize tumor regions. The anti-E-selectin antibody was conjugated on the surface of liposomes, which encapsulated the near-infrared fluorescent substances Cy3 or Cy5.5. The liposomes efficiently recognized human umbilical vein endothelial cells only when E-selectin was induced by angiogenic factors such as TNF-alpha in vitro. Cy5.5 encapsulated into liposomes that were conjugated with an anti-E-selectin antibody successfully visualized Ehrlich ascites tumor cells when transplanted into mice. Thus, E-selectin targeting with liposomes containing a near-infrared fluorescent dye was found effective in visualizing tumors in vivo. This strategy should be extremely useful as a method to identify sentinel lymphatic nodes and angiogenic tumors as well as use for drug delivery to tumor cells.
Asunto(s)
Anticuerpos Monoclonales , Selectina E/inmunología , Inmunoconjugados , Neoplasias/diagnóstico , Animales , Carcinoma de Ehrlich/diagnóstico , Células Endoteliales/citología , Endotelio Vascular , Humanos , Liposomas , Ratones , Neoplasias/patología , Neovascularización Patológica/diagnóstico , Venas UmbilicalesRESUMEN
Molecular targeting agents have become formidable anticancer weapons, which show much promise against the refractory tumors. Functional peptides are among the more desirable of these nanobio-tools. Intracellular delivery of multiple functional peptides forms a basis for potent, non-invasive mode of delivery, providing distinctive therapeutic advantages. Here, we examine growth suppression efficiency of human glioblastomas by dual-peptide targeting. We did simultaneous introduction of two tumor suppressor peptides (p14(ARF) and p16(INK4a) or p16(INK4a) and p21(CIP1) functional peptides) compared with single-peptide introduction using Wr-T-mediated peptide delivery. Wr-T-mediated transport of both p14(ARF) and p16(INK4a) functional peptides (p14-1C and p16-MIS, respectively) into human glioblastoma cell line, U87DeltaEGFR, reversed specific loss of p14 and p16 function, thereby drastically inhibiting tumor growth by >95% within the first 72 h, whereas the growth inhibition was approximately 40% by p14 or p16 single-peptide introduction. Additionally, the combination of p16 and p21(CIP1) (p21-S154A) peptides dramatically suppressed the growth of glioblastoma line Gli36DeltaEGFR, which carries a missense mutation in p53, by >97% after 120 h. Significantly, our murine brain tumor model for dual-peptide delivery showed a substantial average survival enhancement (P < 0.0001) for peptide-treated mice. Wr-T-mediated dual molecular targeting using antitumor peptides is highly effective against growth of aggressive glioblastoma cells in comparison with single molecule targeting. Thus, jointly restoring multiple tumor suppressor functions by Wr-T-peptide delivery represents a powerful approach, with mechanistic implications for development of efficacious molecular targeting therapeutics against intractable human malignancies.
Asunto(s)
Antineoplásicos/farmacología , Glioblastoma/patología , Péptidos/farmacología , Secuencia de Aminoácidos , Animales , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Inhibidor p16 de la Quinasa Dependiente de Ciclina/metabolismo , Ensayos de Selección de Medicamentos Antitumorales , Sinergismo Farmacológico , Femenino , Humanos , Ratones , Ratones Endogámicos BALB C , Ratones Desnudos , Datos de Secuencia Molecular , Mutación/genética , Péptidos/química , Proteína p14ARF Supresora de Tumor/metabolismo , Proteína p53 Supresora de Tumor/metabolismoRESUMEN
We prepared the liposome binding Sialyl Lewis X (SLX) on the surface in order to specifically and efficiently deliver substances (fluorescent materials, chemical substances, proteins, genes, etc.) to inflammation or tumor regions. The liposome with SLX (SLX-Lipo-Cy5.5), in which fluorescent substance Cy5.5 was included, was administered intravenously to arthritis or Ehrlich Ascites Tumor (EAT) bearing mouse, and the accumulation of liposome was observed using two types of in vivo fluorescent imaging equipment. The result was that the accumulation of SLX-Lipo-Cy5.5 to inflammation or tumor regions was significantly higher than the control liposome without sugar chain (Lipo-Cy5.5) at 24 and 48 h after administration. In addition, it was confirmed that this accumulation showed a shift of liposome from blood vessels to the surrounding tissues. Thus, it was proven that this liposome is useful not only as an in vivo bio-imaging reagent but also as a drug delivery system (DDS).