Work

A cell-free TLR5high MSC membrane nanoparticle therapy for Crohn's disease: Targeted immunomodulation via the flagellin/TLR5 axis

Year: 2025

Type: article

Abstract: Inflammatory bowel disease (IBD), including Crohn's disease (CD; Th1/Th17-driven) and ulcerative colitis (Th2-skewed), lacks therapies correcting T-cell imbalance. Current cytokine-focused treatments remain ineffective, while mesenchymal stromal cells (MSCs) therapies are hindered by inherent heterogeneity and challenges related to cell viability maintenance, batch-to-batch consistency, and standardization. This study aimed to (Kaplan, 2015 (1)) identify MSCs subtypes targeting CD pathology, Sebastian and Siegmund (2024) (2) create MSCs-mimicking nanoparticles, and (Li et al., 2016 (3)) propose a “deconstructed cell therapy” framework. Using colon datasets and CD blood samples, Th1/Th17-macrophage dysregulation was mapped. Transcriptomic screening of eight MSCs sources identified dental pulp-derived TLR5 high -MSCs as superior Th1/Th17 inhibitors compared to umbilical cord TLR5 low -MSCs. In colitis models, TLR5 high -MSCs intercepted gut flagellin (Fla), blocking macrophage TLR5/NF-κB to restore T-cell balance. Decellularized MSCs membranes were engineered into nanovesicles (TLR5 high -CMNP), which showed 3.7-fold higher Fla. affinity than antibodies and suppressed Th1/Th17 activity in vitro. In murine colitis, TLR5 high -CMNP achieved comparable efficacy to MSCs (e.g., 68.9 % reduction in MPO scores), while avoiding challenges associated with live-cell administration — such as embolism occurrence (0 % vs. 24 % in MSCs), need for viability maintenance, and potential variability in TLR5 expression. Bioinformatic analysis confirmed TLR5 as pivotal for MSCs specificity, enabling tailored nanoparticle design. This study highlights TLR5 high -CMNP as a safer, cell-free MSCs alternative and introduces a paradigm prioritizing precise immune checkpoint targeting (e.g., Fla./TLR5) over broad cytokine suppression, resolving IBD therapeutic ambiguity through scalable biomimetic nanomaterials. • TLR5 identified as membrane-encoded immune checkpoint governing MSC-mediated Th1/Th17 suppression. • Dental pulp MSCs outperformed eight tissue sources via competitive flagellin-TLR5 interception. • Deconstructed cell therapy framework converts MSC heterogeneity into target-defined nanomaterials. • Biomimetic vesicles show 3.7-fold higher Fla. affinity versus anti-TLR5 antibodies in IBD models.

Source: Journal of Controlled Release

DOI: 10.1016/j.jconrel.2025.114121