DEVELOPMENT OF PRECISION NANOMEDICINE FOR SYSTEMIC LUPUS ERYTHEMATOSUS: EXOSOME-BASED MULTIPLEX DELIVERY OF MORINGIN, QUERCETIN, AND LUTEOLIN

Abstract

Background: Systemic lupus erythematosus (SLE) is a complex autoimmune disorder involving production of autoantibodies, chronic inflammation, dysregulated innate immune signaling, oxidative stress, and progressive renal injury. Combining the effects of several pathogenic mechanisms is a big hurdle for traditional single-target drugs. Extracellular vesicles (EVs) derived from plants (PDEVs) are a novel nanomedicine platform that could be used to deliver multiple bioactive compounds. This work attempted to investigate the effectiveness of a multiplex PDEV formulation comprising morin, quercetin, and luteolin as a multimodal therapeutic approach to experimental SLE. Materials and Methods: A controlled experimental design, consisting of six groups of rats (10 rats per group), was suggested: healthy control, untreated SLE, PDEV-moringin, PDEV-quercetin, PDEV-luteolin, and multiplex PDEV-treated SLE rats. Main systemic, immune, inflammatory, oxidative, and renal characteristics of SLE were replicated in a Pristane-induced lupus-like model. Count the number of diseased rats, and treat the rats during the disease establishment period with the appropriate PDEV formulations. Systemic and renal outcomes were assessed by measuring proteinuria, anti-dsDNA, ANA, complement C3 and C4, and creatinine and BUN. The inflammatory and immunological responses were evaluated by measuring concentrations of TNF-α, IL-6, IL-1β, IL-17A, IFN-α, and BAFF. Molecular analysis involved in the study was TLR7, MyD88, NF-κB p65, NLRP3, and Nrf2, and the oxidative status was evaluated by measuring MDA, SOD, and GSH. The relationship between different disease-associated variables was analysed via correlation and multivariable regression analyses, and overall treatment responses were compared using an integrated PDEV-SLE Precision Therapeutic Index (PDEV SLE-PTI). Results: Untreated SLE Rats showed high levels of proteinuria, anti-dsDNA, ANA, creatinine, BUN, TNF-α, IL 6, IL-1β, IL-17A, IFN-α, and BAFF, while their C3 and C4 levels were decreased. Increased levels of TLR7, MyD88, NF-κB p65, NLRP3, and MDA and decreased levels of Nrf2, SOD, and GSH were also linked to SLE. Single-phytochemical PDEV formulations were all capable of beneficially affecting multiple disease-associated parameters, with multiplex PDEV consistently yielding a treatment-associated response. Multiplex PDEV reduced proteinuria by 74.63%, anti-dsDNA by 69.52%, TNF-α by 61.15%, IFN-α by 69.07%, TLR7 by 64.85%, NF-κB by 64.69%, NLRP3 by 61.91%, and MDA by 57.33%, while producing 76.46% SOD restoration and 72.11% reduction in renal injury. PDEV-SLE-PTI exhibited significant progressive increases from PDEV-moringin through to multiplex PDEV, from 47.22 to 66.90, respectively. The correlation analysis and regression analysis also showed that there were interconnected relationships between renal injury and autoimmunity, inflammatory signaling, activation of NLRP3, and oxidative stress. Conclusion: Multiplex PDEV delivery of moringin, quercetin, and luteolin would be a promising multimodal nanomedicine approach to target the interconnected inflammatory, autoimmune, and oxidative mechanisms in SLE. The superior effect of the multiplex formulation over the individual phytochemical PDEVs indicates that there may be therapeutic advantages of simultaneous modulation of pathways. The current results are preliminary and suggestive. Before clinical translation, further studies will be needed with validated individual-level datasets, dose-response and formal synergy analyses, pharmacokinetic/biodistribution studies, renal targeting studies, and long-term safety evaluation.