ADAM17 but not ADAM10 mediates tumor necrosis factor-alpha and L-selectin shedding from leukocyte membranes.
The release of tumor necrosis factor-alpha (TNF-alpha) from cellular membranes has been shown by different laboratories to be controlled by a disintegrin and metalloprotease, ADAM10 or ADAM17. In contrast, only ADAM17 has shown to be involved in L-selectin shedding. To determine the specific roles of ADAM10 and ADAM17 in the processing of TNF-alpha and L-selectin shedding, antisense oligonucleotides (ASO) targeting both ADAM10 and ADAM17 were identified. We show that ISIS 16337 reduces ADAM17 mRNA and ISIS 100750 reduces ADAM10 mRNA in a sequence-specific and dose-dependent manner in both Jurkat and THP-1 cells. The ADAM17 ASO (ISIS 16337) inhibited both TNF-alpha secretion in THP-1 cells and L-selectin shedding in Jurkat cells, whereas the ADAM10 ASO (ISIS 100750) did not significantly inhibit release of either protein. These results suggest that ADAM17 is one of the major metalloproteases involved in L-selectin shedding as well as TNF-alpha processing. The biologic substrates for ADAM10 in Jurkat and THP-1 cells remain to be elucidated.
Condon
T P
TP
Isis Pharmaceuticals, Carlsbad, CA 92008, USA. tcondon@isisph.com
Flournoy
S
S
Sawyer
G J
GJ
Baker
B F
BF
Kishimoto
T K
TK
Bennett
C F
CF
D051722:ADAM Proteins; D000072198:ADAM17 Protein; D053829:Amyloid Precursor Protein Secretases; D016282:Aspartic Acid Endopeptidases; D002462:Cell Membrane / Q000378:metabolism*; D010450:Endopeptidases / Q000378:metabolism*; D006801:Humans; D019041:L-Selectin / Q000378:metabolism*; D007962:Leukocytes / Q000187:drug effects / Q000378:metabolism*; D008081:Liposomes; D008666:Metalloendopeptidases / Q000378:metabolism; D008845:Microinjections; D016376:Oligonucleotides, Antisense / Q000494:pharmacology; D010714:Phosphatidylethanolamines; D013873:Thionucleotides / Q000494:pharmacology; D014407:Tumor Cells, Cultured; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism*
D008081:Liposomes; D016376:Oligonucleotides, Antisense; D010714:Phosphatidylethanolamines; D013873:Thionucleotides; D014409:Tumor Necrosis Factor-alpha; D019041:L-Selectin; C020888:1,2-dielaidoylphosphatidylethanolamine; D053829:Amyloid Precursor Protein Secretases; D010450:Endopeptidases; D016282:Aspartic Acid Endopeptidases; C509280:BACE1 protein, human; D051722:ADAM Proteins; D008666:Metalloendopeptidases; D000072198:ADAM17 Protein; C000606462:ADAM17 protein, human