Abstract
Drugs that target tumor necrosis factor-alpha (TNF) are particularly important in the treatment of severe inflammatory progression in rheumatoid arthritis, Crohn's disease and psoriasis. Despite the central role of the TNF/TNF receptor (TNFR) in various disease states, there is a paucity of information concerning TNFR2 signaling. In this study, we have developed a simple and highly sensitive cell-death based assay system for analyzing TNFR2-mediated bioactivity that can be used to screen for TNFR2-selective drugs. Using a lentiviral vector, a chimeric receptor was engineered from the extracellular and transmembrane domain of human TNFR2 and the intracellular domain of mouse Fas and the recombinant protein was then expressed in TNFR1(-/-)R2(-/-) mouse preadipocytes. Our results demonstrate that this chimeric receptor is capable of inducing apoptosis by transmembrane- as well as soluble-TNF stimuli. Moreover, we found that our bioassay based on cell death phenotype had an approximately 80-fold higher sensitivity over existing bioassays. We believe our assay system will be an invaluable research tool for studying TNFR2 and for screening TNFR2-targeted drugs.
References
Oct 1, 1992·The Journal of Experimental Medicine·P VandenabeeleW Fiers
May 19, 1995·Cell·B Z StangerB Seed
Feb 1, 1994·Journal of Immunotherapy with Emphasis on Tumor Immunology : Official Journal of the Society for Biological Therapy·K MargolinJ W Mier
Aug 28, 1998·Science·A Ashkenazi, V M Dixit
Jan 29, 1999·Science·H MiyoshiB E Torbett
Dec 12, 2001·Molecular and Cellular Biology·Alicia Algeciras-SchimnichMarcus E Peter
Sep 7, 2002·The Journal of Biological Chemistry·Anja Krippner-HeidenreichPeter Scheurich
Dec 17, 2002·Arthritis and Rheumatism·S Lori BrownM Miles Braun
Mar 26, 2003·Cell Death and Differentiation·H WajantP Scheurich
Jun 28, 2003·Journal of Clinical Oncology : Official Journal of the American Society of Clinical Oncology·Jared A GollobMichael B Atkins
Jul 31, 2003·Cell·Olivier Micheau, Jürg Tschopp
Sep 2, 2003·Nature Reviews. Immunology·Bharat B Aggarwal
Oct 1, 1995·Trends in Cell Biology·P VandenabeeleW Fiers
Jun 11, 2004·Nature·Phil OhJan E Schnitzer
Dec 30, 2004·Clinical Cancer Research : an Official Journal of the American Association for Cancer Research·Hiroko ShibataYasuo Tsutsumi
Dec 19, 2006·Cell·Annalyn GilchristJohn J M Bergeron
Apr 6, 2007·Microbes and Infection·Muazzam JacobsBernhard Ryffel
Citations
May 7, 2011·Biomaterials·Yasuhiro AbeShin-ichi Tsunoda
Jan 5, 2010·Cytokine·Hiroko ShibataYasuo Tsutsumi
Oct 27, 2009·Biomaterials·Yasuo YoshiokaShinsaku Nakagawa
Aug 18, 2009·Biochemical and Biophysical Research Communications·Tetsuya NomuraYasuo Tsutsumi
Dec 17, 2008·Journal of Molecular Biology·Yohei MukaiYasuo Tsutsumi
Dec 19, 2012·Cytokine·Tomohiro MorishigeShinsaku Nakagawa
Feb 27, 2017·The Journal of Biological Chemistry·Masaki InoueShin-Ichi Tsunoda
Jul 3, 2015·Yakugaku zasshi : Journal of the Pharmaceutical Society of Japan·Haruhiko Kamada
May 14, 2020·The Journal of Biological Chemistry·Masaki InoueShin-Ichi Tsunoda