title: Photochemical internalization (PCI) of immunotoxins targeting CD133 is specific and highly potent at femtomolar levels in cells with cancer stem cell properties.
abstract: CD133 is a putative cancer stem cell (CSC) marker for a number of different cancers and is suggested to be a therapeutic target. Since also normal stem cells express CD133 it is of paramount importance that targeting strategies provide a specific and efficient delivery of cytotoxic drugs in only CD133-positive CSCs. In this study, we have employed photochemical internalization (PCI), a minimally invasive method for light-controlled, specific delivery of membrane-impermeable macromolecules from endocytic vesicles to the cytosol, to specifically target CD133-positive cancer cells. We demonstrate that PCI increases the cytotoxic effect of an immunotoxin (IT) targeting CD133-expressing cancer cells of colon (WiDr and HCT116) and pancreas (BxPC-3) origin. The IT consisted of the mAb CD133/1 (AC133) bound to the ribosome inactivating plant toxin saporin (anti-CD133/1-sap). We show that TPCS2a-PCI of anti-CD133/1-sap is specific, and highly cytotoxic at femto-molar concentrations. Specific binding and uptake of CD133/1, was shown by fluorescence microscopy and co-localization with TPCS2a in endosomes/lysosomes was determined by confocal microscopy. CD133(high) WiDr cells, isolated by fluorescence activated cell sorting, had a 7-fold higher capacity to initiate spheroids than CD133(low) cells (P<0.001) and were resistant to photodynamic therapy (PDT). However, PDT-resistance was bypassed by the PCI strategy. Tumor initiation and aggressive growth in athymic nude mice was obtained with only 10 CD133(high) cells in contrast to CD133(low) cells where substantially higher cell numbers were needed. The excellent high efficacy and selectivity of eliminating CD133-expressing cells by PCI warrant further pre-clinical evaluations of this novel therapeutic approach.
authors.affiliation: Department of Radiation Biology, Institute for Cancer Research, Norwegian Radium Hospital, Oslo University Hospital, Oslo, Norway. monica.bostad@rr-research.no
authors.forename: Monica
authors.initials: M
authors.lastname: Bostad
authors.forename: Kristian
authors.initials: K
authors.lastname: Berg
authors.forename: Anders
authors.initials: A
authors.lastname: Høgset
authors.forename: Ellen
authors.initials: E
authors.lastname: Skarpen
authors.forename: Harald
authors.initials: H
authors.lastname: Stenmark
authors.forename: Pål K
authors.initials: PK
authors.lastname: Selbo
mesh_terms: D000071916:AC133 Antigen; D000911:Antibodies, Monoclonal / Q000008:administration & dosage* / Q000737:chemistry; D015703:Antigens, CD / Q000276:immunology*; D045744:Cell Line, Tumor; D016503:Drug Delivery Systems; D006023:Glycoproteins / Q000276:immunology*; D006801:Humans; D014411:Neoplastic Stem Cells / Q000276:immunology*; D010455:Peptides / Q000276:immunology*; D055668:Photochemical Processes; D010778:Photochemotherapy; D017319:Photosensitizing Agents / Q000008:administration & dosage*; D011166:Porphyrins / Q000008:administration & dosage*; D054789:Ribosome Inactivating Proteins, Type 1 / Q000008:administration & dosage* / Q000737:chemistry; D000076985:Saporins
chemical_list: D000071916:AC133 Antigen; D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D006023:Glycoproteins; C000606023:PROM1 protein, human; D010455:Peptides; D017319:Photosensitizing Agents; D011166:Porphyrins; C000606024:Prom1 protein, mouse; D054789:Ribosome Inactivating Proteins, Type 1; C559366:meso-tetraphenyl chlorin disulphonate; D000076985:Saporins