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Updated: Feb 2, 2026
An Adoptive Transfer Model of Rheumatoid Arthritis in Mice
Published on: June 6, 2025
Targeting of Liver Mannan-Binding Lectin-Associated Serine Protease-3 with RNA Interference Ameliorates Disease in a
Nirmal K Banda1, Dhruv Desai2, Robert I Scheinman3
1Division of Rheumatology, Department of Medicine, University of Colorado Anschutz Medical Campus, Aurora, CO 80045.
Abstract:
Mannan-binding lectin-associated serine protease 3 (MASP-3) regulates the alternative pathway of complement and is predominantly synthesized in the liver. The role of liver-derived MASP-3 in the pathogenesis of rheumatoid arthritis (RA) is unknown. We hypothesized that liver-derived MASP-3 is essential for the development of joint damage and that targeted inhibition of MASP-3 in the liver can attenuate arthritis. We used MASP-3-specific small interfering RNAs (siRNAs) conjugated to N-acetylgalactosamine (GalNAc) to specifically target the liver via asialoglycoprotein receptors. Active GalNAc-MASP3-siRNA conjugates were identified, and in vivo silencing of liver MASP-3 mRNA was demonstrated in healthy mice. The s.c. treatment with GalNAc-MASP-3-siRNAs specifically decreased the expression of MASP-3 in the liver and the level of MASP-3 protein in circulation of mice without affecting the levels of the other spliced products. In mice with collagen Ab-induced arthritis, s.c. administration of GalNAc-MASP-3-siRNA decreased the clinical disease activity score to 50% of controls, with decrease in histopathology scores and MASP-3 deposition. To confirm the ability to perform MASP-3 gene silencing in human cells, we generated a lentivirus expressing a short hairpin RNA specific for human MASP-3 mRNA. This procedure not only eliminated the short-term (at day 15) expression of MASP-3 in HepG2 and T98G cell lines but also diminished the long-term (at day 60) synthesis of MASP-3 protein in T98G cells. Our study demonstrates that isoform-specific silencing of MASP-3 in vivo modifies disease activity in a mouse model of RA and suggests that liver-directed MASP3 silencing may be a therapeutic approach in human RA.
Insights
Targeted liver silencing of Mannan-binding lectin-associated serine protease 3 (MASP-3) reduced rheumatoid arthritis symptoms in mice. This approach, using GalNAc-MASP-3-siRNAs, offers a potential new therapy for rheumatoid arthritis.
Area of Science:
- Immunology
- Complement System Biology
- RNA Interference Therapeutics
Background:
- Mannan-binding lectin-associated serine protease 3 (MASP-3) is crucial for the alternative complement pathway and primarily synthesized in the liver.
- The specific role of liver-derived MASP-3 in rheumatoid arthritis (RA) pathogenesis remains largely unexplored.
- This study investigates the hypothesis that liver MASP-3 drives joint damage in RA and that its inhibition could be therapeutic.
Purpose of the Study:
- To investigate the role of liver-derived MASP-3 in rheumatoid arthritis pathogenesis.
- To evaluate the efficacy of targeted liver inhibition of MASP-3 as a potential therapeutic strategy for RA.
- To demonstrate isoform-specific gene silencing of MASP-3 in vivo and in human cell lines.
Main Methods:
- Development and application of N-acetylgalactosamine (GalNAc)-conjugated MASP-3-specific small interfering RNAs (siRNAs) for targeted liver delivery.
- Assessment of MASP-3 mRNA and protein levels in liver and circulation following siRNA treatment in healthy and collagen antibody-induced arthritis (CAIA) mouse models.
- Evaluation of clinical disease activity, joint histopathology, and MASP-3 deposition in arthritic mice treated with GalNAc-MASP-3-siRNAs.
- Validation of MASP-3 gene silencing in human cell lines (HepG2, T98G) using lentiviral short hairpin RNA (shRNA) expression.
Main Results:
- GalNAc-MASP-3-siRNA conjugates effectively and specifically silenced liver MASP-3 mRNA and circulating MASP-3 protein in mice.
- Treatment with GalNAc-MASP-3-siRNAs significantly reduced clinical disease activity (by 50%) and joint pathology in a mouse model of RA.
- MASP-3 deposition in affected joints was also decreased following siRNA treatment.
- Short hairpin RNA targeting human MASP-3 confirmed effective gene silencing in human liver and neuronal cell lines, both short-term and long-term.
Conclusions:
- Isoform-specific in vivo silencing of MASP-3 can modulate disease activity in a murine model of rheumatoid arthritis.
- Targeted, liver-directed MASP-3 gene silencing presents a promising therapeutic avenue for managing human rheumatoid arthritis.
- The study validates the potential of RNA interference strategies for treating complement-mediated inflammatory diseases.
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