Inhibition of Epstein Barr Virus LMP1 gene expression in B lymphocytes by antisense oligonucleotides: uptake and efficacy of lipid-based and receptor-mediated delivery systems.
Epstein Barr Virus (EBV), is associated with an increasing number of lymphoid and epithelial malignancies. Among the genes expressed by EBV during latency, LMP1 plays a key role for growth transformation and immortalization of B lymphocytes. We have previously shown that antisense oligonucleotides (ONs) directed to LMP1 mRNA, effectively suppressed LMP1 gene expression and substantially reduced proliferation of the infected cells. The use of antisense phosphodiester oligonucleotides as therapeutic agents is limited by inefficient cellular uptake and intracellular transport to the target mRNA. We tested the ability of three cationic carriers internalized by different pathways, to increase the delivery of anti-LMP1-ON to their site of action in EBV-infected B lymphocytes. We report here that liposomes, dendrimers or transferrin-polylysine-conjugated ON were internalized by the cells at an extent several fold higher than that of the naked oligomers. However, only the delivery system exploiting the transferrin receptor pathway of internalization, was able to vectorize biologically active antisense LMP1-ON.
Galletti
Roberta
R
Department of Public Health Sciences, University "La Sapienza", P. le A. Moro 5, 00185 Rome, Italy.
Masciarelli
Silvia
S
Conti
Cinzia
C
Matusali
Giulia
G
Di Renzo
Livia
L
Meschini
Stefania
S
Arancia
Giuseppe
G
Mancini
Carlo
C
Mattia
Elena
E
D000818:Animals; D000998:Antiviral Agents / Q000493:pharmacokinetics* / Q000494:pharmacology; D001402:B-Lymphocytes / Q000821:virology*; D002144:Callithrix; D002460:Cell Line; D050091:Dendrimers / Q000493:pharmacokinetics; D004337:Drug Carriers; D015870:Gene Expression / Q000187:drug effects; D004854:Herpesvirus 4, Human / Q000187:drug effects* / Q000235:genetics; D008081:Liposomes / Q000493:pharmacokinetics; D018613:Microscopy, Confocal; D016376:Oligonucleotides, Antisense / Q000493:pharmacokinetics* / Q000494:pharmacology; D011107:Polylysine / Q000031:analogs & derivatives / Q000493:pharmacokinetics; D014168:Transferrin / Q000031:analogs & derivatives / Q000493:pharmacokinetics; D014763:Viral Matrix Proteins / Q000096:biosynthesis / Q000235:genetics
D000998:Antiviral Agents; D050091:Dendrimers; D004337:Drug Carriers; C037662:EBV-associated membrane antigen, Epstein-Barr virus; D008081:Liposomes; D016376:Oligonucleotides, Antisense; D014168:Transferrin; D014763:Viral Matrix Proteins; C072481:transferrin-polylysine conjugate; D011107:Polylysine