title: Light-triggered, efficient cytosolic release of IM7-saporin targeting the putative cancer stem cell marker CD44 by photochemical internalization.
abstract: We have used the site specific and light-depended drug delivery method photochemical internalization (PCI) to release an immunotoxin (IT), targeting the CD44 receptor, into the cytosol of target cells. The IT consisted of a pan CD44 mAb (clone IM7) bound to the ribosome inactivating protein (RIP) saporin by a biotin-streptavidin linker named IM7-saporin. PCI is based upon photosensitizing compounds localized in the membrane of endosomes and lysosomes causing membrane rupture upon illumination followed by release of the IT into the cytosol. In this in vitro study, we have used 7 different human cancer cell lines of various origins to investigate the cytotoxic effect of PCI-based targeting of the cancer stem cell (CSC) marker CD44. Epi-fluorescence microscopy shows both specific binding and uptake of the IM7-Alexa488, after 30 min and 18 h of incubation, and colocalization with the PCI-photosensitizer TPCS2a prior to light-triggered cytosolic release of the CD44-targeting IT. PCI of IM7-saporin resulted in efficient and specific cytotoxicity in CD44-expressing but not in CD44-negative cancer cells. A higher level of reactive oxygen species (ROS) was found in untreated and photodynamic therapy (PDT)-treated LNCaP (CD44(neg)) compared to that of DU145 (CD44(pos)) prostate cancer (PC) cells. This may explain the PDT-resistance observed in the DU145 cells. PCI-based targeting of CD44-expressing cancer cells gives very potent and specific cytotoxic effects and may represent a rational strategy for achieving site-selective elimination of CSCs in aggressive androgen-independent and treatment-resistant PC cells preventing cytotoxic effects on distant normal stem cells.
authors: Department of Radiation Biology, Institute for Cancer Research, Norwegian Radium Hospital, and ‡Cancer Stem Cell Innovation Center (SFI-CAST), Institute for Cancer Research, Norwegian Radium Hospital, Oslo University Hospital , Ullernchausseen 70, 0310 Oslo, Norway.
authors: Monica
authors: M
authors: Bostad
authors: Marianne
authors: M
authors: Kausberg
authors: Anette
authors: A
authors: Weyergang
authors: Cathrine Elisabeth
authors: CE
authors: Olsen
authors: Kristian
authors: K
authors: Berg
authors: Anders
authors: A
authors: Høgset
authors: Pål Kristian
authors: PK
authors: Selbo
mesh_terms: D000911:Antibodies, Monoclonal / Q000737:chemistry; D001710:Biotin / Q000737:chemistry; D045744:Cell Line, Tumor; D003600:Cytosol / Q000378:metabolism; D004337:Drug Carriers / Q000737:chemistry; D011992:Endosomes / Q000378:metabolism; D005434:Flow Cytometry; D006801:Humans; D018960:Hyaluronan Receptors / Q000378:metabolism*; D000922:Immunotoxins / Q000737:chemistry*; D008027:Light; D008247:Lysosomes / Q000378:metabolism; D014411:Neoplastic Stem Cells / Q000187:drug effects*; D010778:Photochemotherapy / Q000379:methods; D017319:Photosensitizing Agents / Q000737:chemistry; D017382:Reactive Oxygen Species / Q000378:metabolism; D054789:Ribosome Inactivating Proteins, Type 1 / Q000737:chemistry*; D000076985:Saporins; D012680:Sensitivity and Specificity; D019809:Streptavidin / Q000737:chemistry; D013997:Time Factors
chemical_list: D000911:Antibodies, Monoclonal; D004337:Drug Carriers; D018960:Hyaluronan Receptors; D000922:Immunotoxins; D017319:Photosensitizing Agents; D017382:Reactive Oxygen Species; D054789:Ribosome Inactivating Proteins, Type 1; D001710:Biotin; D019809:Streptavidin; D000076985:Saporins