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Using RNA-interference to Investigate the Innate Immune Response in Mouse Macrophages
Published on: November 3, 2014
Folate-linked lipid-based nanoparticles deliver a NFkappaB decoy into activated murine macrophage-like RAW264.7 cells
Yoshiyuki Hattori1, Makoto Sakaguchi, Yoshie Maitani
1Institute of Medicinal Chemistry, Hoshi University, Tokyo, Japan.
Abstract:
Activated macrophages are the key effector cells in rheumatoid arthritis (RA) and secrete multiple mediators of inflammation including proinflammatory cytokines. We investigated delivery of a nuclear factor kappa B (NFkappaB) decoy by folate-linked lipid-based nanoparticles (NP-F) into murine macrophages. The expression of folate receptor (FR) in RAW264.7 cells activated by lipopolysaccaride was confirmed by strong expression of FR mRNA, and association of FITC-labeled folate-BSA conjugate. When transfected via NP-F, the NFkappaB decoy was strongly detected in the cytoplasm, and an inhibitory effect on the translocation of NFkappaB into the nucleus was observed at 0.03 microM of the decoy, suggesting that NP-F effectively delivered the NFkappaB decoy into the cytoplasm. This information is of value for the design of NFkappaB decoy carrier systems targeting FR in activated macrophages in gene therapy for autoimmune diseases such as RA.
Insights
This study shows folate-linked nanoparticles effectively deliver nuclear factor kappa B (NFkappaB) decoys into macrophages. This targeted delivery inhibits inflammation, offering a new gene therapy approach for rheumatoid arthritis (RA).
Area of Science:
- Immunology
- Nanotechnology
- Gene Therapy
Background:
- Activated macrophages are central to rheumatoid arthritis (RA) pathogenesis, releasing inflammatory mediators.
- Nuclear factor kappa B (NFkappaB) is a key transcription factor driving inflammation in RA.
Purpose of the Study:
- To investigate the efficacy of folate-linked lipid-based nanoparticles (NP-F) for delivering NFkappaB decoy into murine macrophages.
- To assess the potential of NP-F as a targeted gene therapy carrier for autoimmune diseases.
Main Methods:
- Confirmation of folate receptor (FR) expression on lipopolysaccharide-activated RAW264.7 murine macrophages.
- Transfection of NFkappaB decoy using NP-F and evaluation of its cellular localization and inhibitory effects.
Main Results:
- FR expression was confirmed on activated macrophages via mRNA and folate conjugate binding.
- NP-F successfully delivered NFkappaB decoy into the cytoplasm of macrophages.
- A significant inhibition of NFkappaB nuclear translocation was observed at 0.03 microM decoy concentration.
Conclusions:
- NP-F serves as an effective carrier for delivering NFkappaB decoys into activated macrophages.
- This targeted delivery system holds promise for developing novel gene therapies for RA and other autoimmune diseases.
