title: Development and characterization of immuno-nanocarriers targeting the cancer stem cell marker AC133.
abstract: In the context of targeted therapy, we addressed the possibility of developing a drug delivery nanocarrier capable to specifically reach cancer cells that express the most prominent marker associated with cancer stem cell (CSC) phenotype, AC133. For this purpose, 100nm lipid nanocapsules (LNCs) were functionalized with a monoclonal antibody (mAb) directed against AC133 according to two distinct methods: firstly, post-insertion within 100nm LNCs of a lipid poly(ethylene glycol) functionalized with reactive-sulfhydryl maleimide groups (DSPE-PEG(2000)-maleimide) followed by thiolated mAb coupling, and, secondly, creation of a thiolated lipo-immunoglobulin between DSPE-PEG(2000)-maleimide and AC133, then post-inserted within LNCs. Due to the reduced number of purification steps, lower amounts of DSPE-PEG(2000)-maleimide that were necessary as well as lower number of free maleimide functions present onto the surface of immuno-LNC, the second method was found to be more appropriate. Thus, 126nm AC133-LNC with a zeta potential of -22mV while keeping a narrow distribution were developed. Use of the IgG1κ isotype control-immunoglobulins produced similar control IgG1-LNCs. Micro-Bradford colorimetric assay indicated a fixation of about 40 immunoglobulins per LNC. Use of human Caco-2 cells that constitutively express AC133 (Caco-2-AC133(high)) allowed addressing the behavior of the newly functionalized immuno-LNCs. siRNA knockown strategy permitted to obtain Caco-2-AC133(low) for comparison. Immunofluorescence-combined flow cytometry analysis demonstrated that the epitope-recognition function of AC133 antibody was preserved when present on immuno-LNCs. Although grafting of immunoglobulins onto the surface of LNCs repressed their internalization within Caco-2 cells as evaluated by flow cytometry, AC133-specific cellular binding was obtained with AC133-LNC as assessed by computer-assisted fluorescence microscopy. In conclusion, interest of AC133-LNCs as niche carriers is discussed toward the development of CSC targeted chemo- or radio-nanomedicines.
authors.affiliation: INSERM, U646, F-49933 Angers, France.
authors.forename: E
authors.initials: E
authors.lastname: Bourseau-Guilmain
authors.forename: J
authors.initials: J
authors.lastname: Béjaud
authors.forename: A
authors.initials: A
authors.lastname: Griveau
authors.forename: N
authors.initials: N
authors.lastname: Lautram
authors.forename: F
authors.initials: F
authors.lastname: Hindré
authors.forename: M
authors.initials: M
authors.lastname: Weyland
authors.forename: J P
authors.initials: JP
authors.lastname: Benoit
authors.forename: E
authors.initials: E
authors.lastname: Garcion
mesh_terms: D000071916:AC133 Antigen; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism / Q000494:pharmacology; D000937:Antigen-Antibody Reactions / Q000276:immunology; D015703:Antigens, CD / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D000951:Antigens, Neoplasm / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D018938:Caco-2 Cells; D002448:Cell Adhesion / Q000187:drug effects; D002462:Cell Membrane / Q000378:metabolism; D003593:Cytoplasm / Q000378:metabolism; D016503:Drug Delivery Systems / Q000379:methods*; D004705:Endocytosis / Q000502:physiology; D006023:Glycoproteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D006801:Humans; D007096:Imidoesters / Q000737:chemistry; D054709:Lecithins / Q000737:chemistry; D008301:Maleimides / Q000737:chemistry; D053769:Nanocapsules / Q000737:chemistry*; D058726:Nanoconjugates / Q000737:chemistry*; D010316:Particle Size; D010455:Peptides / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D010714:Phosphatidylethanolamines / Q000737:chemistry; D011092:Polyethylene Glycols / Q000737:chemistry; D034741:RNA, Small Interfering / Q000235:genetics; D055672:Static Electricity; D013229:Stearic Acids / Q000737:chemistry; D013438:Sulfhydryl Compounds / Q000138:chemical synthesis; D013499:Surface Properties; D014280:Triglycerides / Q000737:chemistry
chemical_list: C519184:1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-methoxy-poly(ethylene glycol 2000); D000071916:AC133 Antigen; D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D000951:Antigens, Neoplasm; D006023:Glycoproteins; D007096:Imidoesters; D054709:Lecithins; D008301:Maleimides; D053769:Nanocapsules; D058726:Nanoconjugates; C000606023:PROM1 protein, human; D010455:Peptides; D010714:Phosphatidylethanolamines; D034741:RNA, Small Interfering; D013229:Stearic Acids; D013438:Sulfhydryl Compounds; D014280:Triglycerides; D011092:Polyethylene Glycols; C067028:Solutol HS 15; C005905:methyl 4-mercaptobutyrimidate