title: Death receptor 3 mediates TNFSF15- and TNFα-induced endothelial cell apoptosis.
abstract: Tumor necrosis factor superfamily 15 (TNFSF15) suppresses angiogenesis by specifically inducing apoptosis in proliferating endothelial cells. Death receptor 3 (DR3), a member of the TNF receptor superfamily (TNFRSF25), has been identified as a receptor for TNFSF15 to activate T cells. It is unclear, however, whether DR3 mediates TNFSF15 activity on endothelial cells. Here we show that siRNA-mediated knockdown of DR3 in an in vivo Matrigel angiogenesis assay, or in adult bovine aortic endothelial (ABAE) cell cultures, leads to resistance of endothelial cells to TNFSF15-induced apoptosis. Interestingly, DR3-depleted cells also exhibited markedly diminished responsiveness to TNFα cytotoxicity, even though DR3 is not a receptor for TNFα. Treatment of the cells with either TNFSF15 siRNA or a TNFSF15-neutralizing antibody, 4-3H, also results in a significant inhibition of TNFα-induced apoptosis. Mechanistically, DR3 siRNA treatment gives rise to an increase of ERK1/2 MAPK activity, and up-regulation of the anti-apoptotic proteins c-FLIP and Bcl-2, thus strengthening apoptosis-resisting potential in the cells. These findings indicate that DR3 mediates TNFSF15-induced endothelial cell apoptosis, and that up-regulation of TNFSF15 expression stimulated by TNFα is partly but significantly responsible for TNFα-induced apoptosis in endothelial cells.
authors.affiliation: State Key Laboratory of Medicinal Chemical Biology and College of Pharmacy, Nankai University, Tianjin Key Laboratory of Molecular Drug Research, Tianjin, China; Collaborative Innovation Center for Biotherapy, Nankai University, China.
authors.forename: Li-Xia
authors.initials: LX
authors.lastname: Xu
authors.affiliation: USC Norris Cancer Hospital, Department of Medicine, University of Southern California, 1441 Eastlake Avenue, NOR 6330, Los Angeles, CA 90033, United States.
authors.forename: Sammy
authors.initials: S
authors.lastname: Grimaldo
authors.affiliation: State Key Laboratory of Medicinal Chemical Biology and College of Pharmacy, Nankai University, Tianjin Key Laboratory of Molecular Drug Research, Tianjin, China; Collaborative Innovation Center for Biotherapy, Nankai University, China.
authors.forename: Jian-Wei
authors.initials: JW
authors.lastname: Qi
authors.affiliation: State Key Laboratory of Medicinal Chemical Biology and College of Pharmacy, Nankai University, Tianjin Key Laboratory of Molecular Drug Research, Tianjin, China; Collaborative Innovation Center for Biotherapy, Nankai University, China.
authors.forename: Gui-Li
authors.initials: GL
authors.lastname: Yang
authors.affiliation: State Key Laboratory of Medicinal Chemical Biology and College of Pharmacy, Nankai University, Tianjin Key Laboratory of Molecular Drug Research, Tianjin, China; Collaborative Innovation Center for Biotherapy, Nankai University, China.
authors.forename: Ting-Ting
authors.initials: TT
authors.lastname: Qin
authors.affiliation: State Key Laboratory of Medicinal Chemical Biology and College of Pharmacy, Nankai University, Tianjin Key Laboratory of Molecular Drug Research, Tianjin, China; Collaborative Innovation Center for Biotherapy, Nankai University, China.
authors.forename: Huai-Yuan
authors.initials: HY
authors.lastname: Xiao
authors.affiliation: Collaborative Innovation Center for Biotherapy, Nankai University, China; Nankai University School of Medicine, Tianjin, China.
authors.forename: Rong
authors.initials: R
authors.lastname: Xiang
authors.affiliation: Collaborative Innovation Center for Biotherapy, Nankai University, China; Sichuan University College of Life Sciences, Chengdu, China.
authors.forename: Zhixiong
authors.initials: Z
authors.lastname: Xiao
authors.affiliation: State Key Laboratory of Medicinal Chemical Biology and College of Pharmacy, Nankai University, Tianjin Key Laboratory of Molecular Drug Research, Tianjin, China; Collaborative Innovation Center for Biotherapy, Nankai University, China. Electronic address: liluyuan@nankai.edu.cn.
authors.forename: Lu-Yuan
authors.initials: LY
authors.lastname: Li
authors.affiliation: State Key Laboratory of Medicinal Chemical Biology and College of Pharmacy, Nankai University, Tianjin Key Laboratory of Molecular Drug Research, Tianjin, China; Collaborative Innovation Center for Biotherapy, Nankai University, China. Electronic address: zzs@nankai.edu.cn.
authors.forename: Zhi-Song
authors.initials: ZS
authors.lastname: Zhang
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D015153:Blotting, Western; D053446:CASP8 and FADD-Like Apoptosis Regulating Protein / Q000378:metabolism; D020169:Caspases / Q000378:metabolism; D002417:Cattle; D002460:Cell Line; D004317:Doxorubicin / Q000494:pharmacology; D042783:Endothelial Cells / Q000187:drug effects* / Q000378:metabolism / Q000502:physiology; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism; D005260:Female; D015870:Gene Expression; D008810:Mice, Inbred C57BL; D018919:Neovascularization, Physiologic / Q000187:drug effects; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism; D034622:RNA Interference; D053219:Receptors, Tumor Necrosis Factor, Member 25 / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism / Q000494:pharmacology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D053380:Tumor Necrosis Factor Ligand Superfamily Member 15 / Q000235:genetics / Q000494:pharmacology*; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*
chemical_list: D053446:CASP8 and FADD-Like Apoptosis Regulating Protein; C506387:Cflar protein, mouse; D019253:Proto-Oncogene Proteins c-bcl-2; D053219:Receptors, Tumor Necrosis Factor, Member 25; D011994:Recombinant Proteins; C505945:Tnfsf15 protein, rat; D053380:Tumor Necrosis Factor Ligand Superfamily Member 15; D014409:Tumor Necrosis Factor-alpha; D004317:Doxorubicin; D048049:Extracellular Signal-Regulated MAP Kinases; D020169:Caspases
keywords: Angiogenesis; Apoptosis; Autocrine; Cytokine; Death receptor