title: Noninvasive imaging of apoptosis induced by adenovirus-mediated cancer gene therapy using a caspase-3 biosensor in living subjects.
abstract: We attempted to visualize the serial induction of caspase-3-dependent apoptosis mediated by Fas ligand/tumor necrosis factor-related apoptosis-inducing ligand (FasL/TRAIL) adenoviral gene therapy in mice bearing human glioma xenografts using a caspase-3 biosensor and monitored its therapeutic effects. Human D54 glioma cells expressing both the caspase-3 sensor and the Renilla luciferase (Rluc) gene were established (referred to as D54-CR cells). The bioluminescence imaging (BLI) signals of the caspase-3 sensor in the D54-CR cells were increased in a time- and virus dose-dependent manner by Ad-TRAIL or Ad-FasL transduction. Fluorescence-activated cell sorting (FACS) analysis revealed an increase in both cleaved caspase-3 or poly(ADP-ribose) polymerase (PARP) and annexin V- and propidium iodide-positive cells depending on the dosage of administered virus. Ad-FasL treatment resulted in a significant increase in the BLI activity of the caspase-3 sensor in the D54-CR tumors, which were ≈ 8.2, ≈ 12.9, and ≈ 46.6 times higher than those of control at 12 hours, 24 hours, and 96 hours posttreatment, respectively. In contrast, a significant reduction in Rluc activity, as a surrogate marker of cell viability, was detected in the tumors treated with Ad-FasL but not in those treated with Ad-null. Overall, the activation of caspase-3-dependent apoptosis induced by Ad-FasL/Ad-TRAIL gene therapy was successfully monitored by a sensitive imaging platform for caspase-3 activation.
authors.forename: Thoudam Debraj
authors.initials: TD
authors.lastname: Singh
authors.forename: Ho Won
authors.initials: HW
authors.lastname: Lee
authors.forename: Sang-Woo
authors.initials: SW
authors.lastname: Lee
authors.forename: Jeoung-Hee
authors.initials: JH
authors.lastname: Ha
authors.forename: Alnawaz
authors.initials: A
authors.lastname: Rehemtulla
authors.forename: Byeong-Cheol
authors.initials: BC
authors.lastname: Ahn
authors.forename: Young Hyun
authors.initials: YH
authors.lastname: Jeon
authors.forename: Jaetae
authors.initials: J
authors.lastname: Lee
mesh_terms: D000256:Adenoviridae / Q000235:genetics* / Q000378:metabolism; D000818:Animals; D017209:Apoptosis; D015374:Biosensing Techniques; D053148:Caspase 3 / Q000378:metabolism*; D045744:Cell Line, Tumor; D053222:Fas Ligand Protein / Q000235:genetics / Q000378:metabolism; D015316:Genetic Therapy; D005822:Genetic Vectors / Q000008:administration & dosage; D005910:Glioma / Q000000981:diagnostic imaging* / Q000628:therapy*; D006801:Humans; D049410:Luciferases, Renilla; D049408:Luminescent Agents; D051379:Mice; D008807:Mice, Inbred BALB C; D009374:Neoplasms, Experimental; D011877:Radionuclide Imaging
chemical_list: C505661:FASLG protein, human; D053222:Fas Ligand Protein; D049408:Luminescent Agents; D049410:Luciferases, Renilla; D053148:Caspase 3