Suppression of human monocyte tissue factor synthesis by antisense oligodeoxynucleotide

Area of Science:

• Biochemistry
• Molecular Biology
• Immunology

Background:

• Tissue factor (TF) initiates the blood coagulation cascade, making its cell biology crucial for understanding various disorders.
• Inhibiting TF synthesis is a potential therapeutic strategy for TF-related pathologies.

Purpose of the Study:

• To investigate the feasibility of inhibiting human blood monocyte TF synthesis using an antisense oligodeoxynucleotide (aODN).
• To explore the use of CD14 receptor-mediated endocytosis for delivering TF aODN to monocytes.

Main Methods:

• An antisense oligodeoxynucleotide (aODN) targeting human monocyte TF mRNA was designed.
• A DNA carrier system using anti-CD14 antibody linked to poly(L-lysine) facilitated CD14 receptor-mediated endocytosis for aODN delivery.
• TF activity was measured in monocytes treated with TF aODN and endotoxin, with control experiments using sense ODN, mismatched aODN, and irrelevant aODN.

Main Results:

• Co-treatment of monocytes with TF aODN and endotoxin resulted in significant suppression of TF activity (80.4 +/- 2.2%).
• Control experiments confirmed the specificity of the TF aODN, excluding non-specific inhibitory effects.
• Cytotoxicity of the DNA carrier complex was evaluated.

Conclusions:

• The TF mRNA antisense ODN specifically suppressed the synthesis of biologically active monocyte TF.
• Antisense ODNs represent a promising tool for investigating monocyte/macrophage TF function in health and disease.