BACKGROUND: To establish new strategies for the treatment of proliferative vitreoretinopathy (PVR), we investigated new members of a recently discovered apoptosis-inducing receptor-ligand system in human retinal pigment epithelial (RPE) cells. TRAIL (Apo2-L) and Apo3-L are capable of inducing cell death via their receptors Trail-R1 to Trail-R4 and TRAMP. The goal of this study was to prove the existence of these new apoptosis-inducing receptors and ligands in RPE cells. METHODS: Human RPE cells, cultured or prepared directly from the eye, were examined by RT-PCR. Immunohistochemistry of epiretinal membranes of traumatic PVR was performed for the detection of TRAIL and Trail-R1. Protein expression of Trail-R1 was examined in cultured human RPE cells by western blot. Cell death after TRAIL treatment of human RPE cells was measured by crystal violet staining. RESULTS: For RPE cells derived directly from the eye, we detected mRNAs of Trail-R2, Trail-R3, TRAIL, and APO3-L, but not Trail-R1, Trail-R4, and TRAMP. All the examined transcripts were detected in human P0 RPE cells in vitro. Immunohistochemical studies on PVR membranes identified TRAIL and Trail-R1. Western blot confirmed the presence of Trail-R1 in cultured human RPE cells. TRAIL failed to kill RPE cells in vitro, but showed a strong synergistic killing effect when coincubated with protein (cycloheximide) or RNA (actinomycin D) synthesis inhibitor. CONCLUSIONS: We detected a novel apoptosis-inducing receptor-ligand system in RPE cells. An induction of apoptosis as a treatment of PVR seems to be possible. Further investigations are needed including an animal model of PVR.
BACKGROUND: To establish new strategies for the treatment of proliferative vitreoretinopathy (PVR), we investigated new members of a recently discovered apoptosis-inducing receptor-ligand system in humanretinal pigment epithelial (RPE) cells. TRAIL (Apo2-L) and Apo3-L are capable of inducing cell death via their receptors Trail-R1 to Trail-R4 and TRAMP. The goal of this study was to prove the existence of these new apoptosis-inducing receptors and ligands in RPE cells. METHODS:Human RPE cells, cultured or prepared directly from the eye, were examined by RT-PCR. Immunohistochemistry of epiretinal membranes of traumatic PVR was performed for the detection of TRAIL and Trail-R1. Protein expression of Trail-R1 was examined in cultured human RPE cells by western blot. Cell death after TRAIL treatment of human RPE cells was measured by crystal violet staining. RESULTS: For RPE cells derived directly from the eye, we detected mRNAs of Trail-R2, Trail-R3, TRAIL, and APO3-L, but not Trail-R1, Trail-R4, and TRAMP. All the examined transcripts were detected in human P0 RPE cells in vitro. Immunohistochemical studies on PVR membranes identified TRAIL and Trail-R1. Western blot confirmed the presence of Trail-R1 in cultured human RPE cells. TRAIL failed to kill RPE cells in vitro, but showed a strong synergistic killing effect when coincubated with protein (cycloheximide) or RNA (actinomycin D) synthesis inhibitor. CONCLUSIONS: We detected a novel apoptosis-inducing receptor-ligand system in RPE cells. An induction of apoptosis as a treatment of PVR seems to be possible. Further investigations are needed including an animal model of PVR.
Authors: Christina L Zuch de Zafra; Avi Ashkenazi; Walter C Darbonne; Melissa Cheu; Klara Totpal; Shirley Ortega; Heather Flores; Mark D Walker; Bruce Kabakoff; Bert L Lum; Barbara J Mounho-Zamora; Scot A Marsters; Noël O Dybdal Journal: Cell Death Dis Date: 2016-08-11 Impact factor: 8.469