title: Novel antisense therapeutics delivery systems: In vitro and in vivo studies of liposomes targeted with anti-CD20 antibody.
abstract: Antisense gene therapy using molecules such as antisense oligodeoxynucleotides, siRNA or miRNA is a very promising strategy for the treatment of neoplastic diseases. It can be combined with other treatment strategies to enhance therapeutic effect. In acute leukemias, overexpression of the antiapoptotic gene BCL2 is observed in more than 70% of cases. Therefore, reduction of the Bcl-2 protein level could, in itself, prevent the development of cancer or could possibly help sensitize cancer cells to apoptosis inducers. The main objective of our work is to develop therapeutic liposome formulations characterized by high transfection efficiency, stability in the presence of serum, as well as specificity and toxicity for target (leukemic) cells. Each of our liposomal formulations consists of a core composed of antisense oligonucleotides complexed by either cationic lipid, DOTAP, or a synthetic polycation, polyethyleneimine, encapsulated within liposomes modified with polyethylenoglycol. In addition, the liposomal shells are enriched with covalently-bound antibodies recognizing a well characterized bio-marker, CD20, exposed on the surface of leukemia cells. The resulting immunoliposomes selectively and effectively reduced the expression of BCL2 in target cells. Model animal experiments carried out on mice-engrafted tumors expressing the specific marker showed high efficiency of the liposome formulations against specific tumor development. In conclusion, we show that lipid formulations based on a polyplex or lipoplex backbone additionally equipped with antibodies are promising non-viral vectors for specific oligonucleotide transfer into human tumor cells.
authors.affiliation: Laboratory of Cytobiochemistry, Biotechnology Faculty, University of Wrocław, Joliot-Curie 14a, 50-383 Wrocław, Poland; Electron Microscopy Laboratory, Faculty of Biology, University of Environmental and Life Sciences Wroclaw, Kożuchowska 5b, 50-631 Wroclaw, Poland.
authors.forename: Justyna M
authors.initials: JM
authors.lastname: Meissner
authors.affiliation: Laboratory of Cytobiochemistry, Biotechnology Faculty, University of Wrocław, Joliot-Curie 14a, 50-383 Wrocław, Poland.
authors.forename: Monika
authors.initials: M
authors.lastname: Toporkiewicz
authors.affiliation: Laboratory of Cytobiochemistry, Biotechnology Faculty, University of Wrocław, Joliot-Curie 14a, 50-383 Wrocław, Poland.
authors.forename: Aleksander
authors.initials: A
authors.lastname: Czogalla
authors.affiliation: Laboratory of Cytobiochemistry, Biotechnology Faculty, University of Wrocław, Joliot-Curie 14a, 50-383 Wrocław, Poland.
authors.forename: Lucyna
authors.initials: L
authors.lastname: Matusewicz
authors.affiliation: Wrocław Medical University, Department and Clinic of Haematology, Blood Neoplasms, and Bone Marrow Transplantation ul. L, Pasteura 4, 50-367 Wroclaw, Poland.
authors.forename: Kazimierz
authors.initials: K
authors.lastname: Kuliczkowski
authors.affiliation: Laboratory of Cytobiochemistry, Biotechnology Faculty, University of Wrocław, Joliot-Curie 14a, 50-383 Wrocław, Poland. Electronic address: afsbc@ibmb.uni.wroc.pl.
authors.forename: Aleksander F
authors.initials: AF
authors.lastname: Sikorski
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000008:administration & dosage* / Q000737:chemistry / Q000493:pharmacokinetics; D018951:Antigens, CD20 / Q000276:immunology / Q000378:metabolism; D017209:Apoptosis; D002051:Burkitt Lymphoma / Q000235:genetics / Q000276:immunology / Q000378:metabolism / Q000473:pathology / Q000628:therapy*; D045744:Cell Line, Tumor; D005229:Fatty Acids, Monounsaturated / Q000737:chemistry; D015972:Gene Expression Regulation, Neoplastic; D015316:Genetic Therapy / Q000379:methods*; D006801:Humans; D018796:Immunoconjugates / Q000008:administration & dosage* / Q000737:chemistry / Q000493:pharmacokinetics; D008055:Lipids / Q000737:chemistry*; D008081:Liposomes; D008297:Male; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D016376:Oligonucleotides, Antisense / Q000008:administration & dosage* / Q000235:genetics / Q000493:pharmacokinetics; D011092:Polyethylene Glycols / Q000737:chemistry; D011094:Polyethyleneimine / Q000737:chemistry; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000235:genetics / Q000378:metabolism; D000644:Quaternary Ammonium Compounds / Q000737:chemistry; D014018:Tissue Distribution; D014162:Transfection / Q000379:methods*; D023041:Xenograft Model Antitumor Assays
chemical_list: D000911:Antibodies, Monoclonal; D018951:Antigens, CD20; C000595707:BCL2 protein, human; D005229:Fatty Acids, Monounsaturated; D018796:Immunoconjugates; D008055:Lipids; D008081:Liposomes; D016376:Oligonucleotides, Antisense; D019253:Proto-Oncogene Proteins c-bcl-2; D000644:Quaternary Ammonium Compounds; D011092:Polyethylene Glycols; D011094:Polyethyleneimine; C070046:1,2-dioleoyloxy-3-(trimethylammonium)propane
keywords: Antisense nucleotide therapy; B-cell malignancies; BCL2 gene; Bcl-2 protein; Cationic lipids; Polyethylenimine; Targeted liposomes