Chemically modified ribozyme targeting TNF-alpha mRNA regulates TNF-alpha and IL-6 synthesis in synovial fibroblasts of patients with rheumatoid arthritis.
Rheumatoid arthritis (RA) is chronic polyarthritis in which a variety of inflammatory cytokines play a role. Since tumor necrosis factor-alpha (TNF-alpha) is one of the most important cytokines in the pathogenesis of RA, we evaluated the feasibility of ribozymes as a therapeutic agent to control the inflammatory process of RA synovium. A hammerhead ribozyme against TNF-alpha was chemically modified to increase nuclease resistance and added to RA fibroblastlike cell cultures without using a delivery system. The cellular uptake of fluorescent-labeled ribozyme into synovial cells was found to last at least 48 hr by confocal laser scanning microscopy. The ribozyme targeting TNF-alpha gene inhibited both the expression of TNF-alpha mRNA and the secretion of TNF-alpha and IL-6. The cytotoxic effect by the ribozyme on synovial cells was negligible when determined by an alamar blue assay. Chemically modified ribozymes designed to suppress the TNF-alpha gene may be potential as a therapeutic agent for rheumatoid arthritis.
Takahashi
Minako
M
Department of Clinical Medicine, Tohoku University, School of Medicine, Sendai, Japan. minakot@mail.cc.tohoku.ac.jp
Funato
Tadao
T
Suzuki
Yoko
Y
Fujii
Hiroshi
H
Ishii
Keiko Kumura
KK
Kaku
Mitsuo
M
Sasaki
Takeshi
T
D001172:Arthritis, Rheumatoid / Q000188:drug therapy* / Q000235:genetics / Q000276:immunology*; D001483:Base Sequence; D002478:Cells, Cultured; D005347:Fibroblasts / Q000187:drug effects / Q000276:immunology; D006801:Humans; D066298:In Vitro Techniques; D015850:Interleukin-6 / Q000096:biosynthesis*; D016337:RNA, Catalytic / Q000737:chemistry / Q000235:genetics / Q000627:therapeutic use*; D012333:RNA, Messenger / Q000235:genetics* / Q000378:metabolism*; D013583:Synovial Membrane / Q000187:drug effects / Q000276:immunology; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis* / Q000235:genetics*
D015850:Interleukin-6; D016337:RNA, Catalytic; D012333:RNA, Messenger; D014409:Tumor Necrosis Factor-alpha