Title : Engineered MSC-EVs for targeted PAD4 miRNA delivery to control NETosis in neonatal viral sepsis
Abstract:
Rationale: Neonatal viral sepsis remains a major clinical challenge, especially in resource-limited settings, due to high mortality and the lack of effective antiviral treatments. A key pathogenic feature of viral sepsis is excessive Neutrophil Extracellular Trap (NET) formation (NETosis), which, while initially protective, can drive inflammation and tissue damage. PAD4 (Peptidyl Arginine Deiminase 4) is a critical enzyme in NETosis, catalyzing histone citrullination and chromatin decondensation. Targeting PAD4 offers a promising therapeutic strategy, but conventional inhibitors carry risks of off-target effects and immune suppression. This study proposes using engineered Mesenchymal Stem Cell-derived Extracellular Vesicles (MSC-EVs) loaded with PAD4-targeting miRNAs to specifically inhibit NETosis, offering a novel, biocompatible, and targeted approach to neonatal viral sepsis.
Objectives: Dentify and validate PAD4-targeting miRNAs (e.g. miR-155, miR-223, miR-3164). Engineer MSCs to produce EVs enriched with PAD4-specific miRNAs. Characterize the MSC-EVs for efficient cargo loading, targeting ability, and immune modulation. Evaluate the efficacy of these EVs in suppressing NETosis in vitro and in in vivo models of viral-induced neonatal sepsis. Assess the safety, biodistribution, and long-term therapeutic outcomes of the treatment.
Hypothesis: Engineered MSC-derived EVs carrying PAD4-targeting miRNAs can selectively suppress NETosis in viral neonatal sepsis by downregulating PAD4, thereby reducing inflammation and tissue damage without compromising essential immune responses.
Significance to the study: This project represents a pioneering application of cell-free, EV-based RNA therapeutics for neonatal viral sepsis. If successful, it could: Uncover new insights into miRNA-mediated regulation of NETosis. Establish MSC-EVs as a safe and efficient delivery platform for targeted immune modulation in neonates. Lay the groundwork for next generation antiviral immunotherapies that are non-antibiotic, personalized, and safer for vulnerable neonatal populations.
Keywords: MSC Extracellular Vesicles, Engineered EVs, RNA Delivery, PAD4-Targeting miRNA, siRNA Therapy, Targeted Therapy
