Efficient Treatment of Rheumatoid Arthritis by Degradable LPCE Nano-Conjugate-Delivered p65 siRNA

Abstract

Rheumatoid arthritis (RA) is one of the most common autoimmune diseases worldwide, causing severe cartilage damage and disability. Despite the recent progress made in RA treatment, limitations remain in achieving early and efficient therapeutic intervention. Advanced therapeutic strategies are in high demand, and siRNA-based therapeutic technology with a gene-silencing ability represents a new approach for RA treatment. In this study, we created a cationic delivery micelle consisting of low-molecular-weight (LMW) polyethylenimine (PEI)–cholesterol–polyethylene glycol (PEG) (LPCE) for small interfering RNA (siRNA)-based RA gene therapy. The carrier is based on LMW PEI and modified with cholesterol and PEG. With these two modifications, the LPCE micelle becomes multifunctional, and it efficiently delivered siRNA to macrophages with a high efficiency greater than 70%. The synthesized LPCE exhibits strong siRNA protection ability and high safety. By delivering nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) p65 siRNA, the p65 siRNA/LPCE complex efficiently inhibited macrophage-based cytokine release in vitro. Local administration of the p65 siRNA/LPCE complex exhibited a fast and potent anti-inflammatory effect against RA in a mouse model. According to the results of this study, the functionalized LPCE micelle that we prepared has potential gene therapeutic implications for RA.

siRNA‐based nanotherapeutics as emerging modalities for immune‐mediated diseases: A preliminary review

“…For knocking down specific genes, siRNAs have shown promise, but their application as therapeutic agents has limits due to their hydrophilicity and ease of degradation. As a result, most cell types can be TNF-α-siRNA NPs Co-delivery of TNF-α siRNA and TPPS2a, as an encouraging topical therapy against psoriasis (Suzuki et al, 2021) siRNA-LNPs Distinguished reduction of IRF8 mRNA exhibited a targeted immunomodulatory effect ex vivo and in vivo in the DSS colitis model (Veiga, Goldsmith et al, 2019) siRNA-polymeric micelles Accumulation in the inflamed large intestine, decreased inflammatory reactions in UC model mice, demonstrating a positive inhibitory effect (Ibaraki et al, 2022) NF-κB p65-siRNA multifunctionalized polymeric micelles Potential tactic for gene therapy against RA (Chen et al, 2022) Bio-reducible PbAE NPs Significant results against human GBM cells through inhibition and migration of inflammatory cancer cells (Kozielski, Ruiz-Valls et al, 2019) Au-LNHy coated with AuNPs and comodified with PEG and α8 integrin, then loaded with dexamethasone and TGFβ1 siRNA…”

“…Moreover, NF‐κB p65 significantly contributed to the pathophysiological proinflammatory cascades during RA. So, it is noteworthy to confirm that silencing the expression of NF‐κB p65 in macrophages through siRNA‐based nano‐therapeutics can be a potential tactic for gene therapy against RA (Chen, Zhou et al, 2022).…”

“…Xiao Hua Chen and coworkers (2022) developed a novel multifunctionalized LMW PEI–cholesterol–PEG (LPCE) delivery micelle against RA gene therapy. Furthermore, NF‐κB p65 siRNA was conjugated to the cationic polymeric micelles for regulating inflammation, immunity, cell proliferation, and cellular apoptosis for the targeted and safest delivery.…”

“…Moreover, NF-κB p65significantly contributed to the pathophysiological proinflammatory cascades during RA. So, it is noteworthy to confirm that silencing the expression of NF-κB p65 in macrophages through siRNA-based nano-therapeutics can be a potential tactic for gene therapy against RA(Chen, Zhou et al, 2022).In another related research, Scheinman and associates developed RGD functionalized poly(lactide-co-glycolytic) acid (PLGA)-based NPs to deliver STAT1 siRNA, as RGD peptide is linked with the propagation of siRNA in cellular tissues. Therefore, RGD peptide was synthesized in preparing NPs for tracking via siRNA.…”

The emerging potential of siRNA nanotherapeutics in treatment of arthritis

“… [144] PEGylation siRNA incorporated PLGA nanoparticles TNF-α siRNA Mcl-1 siRNA Particle size: 192 ± 12 nm, Zeta potential: −26.7 ± 2.3 mV CIA Intravenously injected Inhibited macrophage-based cytokine release and anti-inflammatory effect. [145] CH: Chitosan; CIA: collagen-induced arthritis; MCL-1: myeloid cell leukemia-I; COX2: Cyclooxygenase-2; iNOS: inducible nitric oxide; Ki-67: Kiel-67 (monoclonal antibody); SAINT-C18: Cationic lipid 1-methyl-4-(cis-9-dioleyl)methyl-pyridinium-chloride; VCAM-1: Vascular cell adhesion molecule 1; tGC: Thiolated glycol chitosan; PDAPEI: Dopamine and PEI copolymerized nanodots; PCL: Poly-ε-caprolactone; DEAE: Diethylethylamine; CAIA: Collagen-Antibody Induced Arthritis; MTX: Methotrexate; AIA: Adjuvant-induced arthritis; BAFF-R: B-cell activating factor receptor; HA: Hyaluronic acid; LPCE:low-molecular-weight-polyethylenimine-cholesterol–polyethylene-glycol; PCADK: poly (cyclohexane-1,4-diyl acetone dimethylene ketal); SWCNTs: single-walled carbon nanotubes. …”