MicroRNA-200a as a therapeutic agent for acute and chronic liver pathologies and regeneration.

Abstract

Effective therapies targeting the molecular pathogenesis of liver diseases remain an unmet clinical need. Here, using acute and chronic liver pathology models, we demonstrate a dual therapeutic role for miR-200a encapsulated in poly(lactic acid) (PLA) nanoparticles (NPs) in both treating liver pathology and stimulating regeneration. This rationale is based on miR-200a's central role in regulating oxidative and endoplasmic reticulum (ER) stress, key drivers of liver diseases, enhancing liver regeneration and preventing disease development. Herein, we report the design and therapeutic validation of a nanomedicine platform comprising biocompatible PLA NPs for the hepatic delivery of miR-200a. The resulting PLA@miR-200a NPs (155 ± 39 nm in diameter) demonstrated successful cargo encapsulation and were internalized by hepatocytes in ex vivo liver slice models with minimal cytotoxicity. Pre-complexation of the miR with spermidine facilitated efficient encapsulation within the PLA matrix. Intracellular kinetic studies revealed a degradation-mediated, sustained release profile of the miR payload over 48 h. Crucially, the functional efficacy of this delivery mechanism was validated in vivo, where PLA@miR-200a successfully activated the Keap1/Nrf2 signaling pathway in target hepatocytes, achieving a reliable physiological effect. In a murine model of acute acetaminophen-induced liver injury, a single intraperitoneal administration of PLA@miR-200a profoundly mitigated hepatotoxicity, reduced dystrophic changes, normalized liver enzyme levels, and restored hepatocyte proliferation. Advanced metabolic FLIM imaging in combination with molecular analysis revealed revitalized of cellular biosynthetic activity and antioxidant capacity. Furthermore, in a chronic carbon tetrachloride-induced fibrosis model, the therapy demonstrated potent anti-fibrotic activity, reducing collagen deposition by two stages on the Metavir scale and promoting the resolution of fibrotic septa. In addition, the liver's regenerative capacity and metabolic state were normalized, as evidenced by restored hepatocyte proliferation and FLIM-based metabolic imaging. This work establishes that miR-200a-based nanotherapy has a high potential as a universal, dual-action strategy for not only treating a wide range of liver pathologies but also enhancing the organ's intrinsic regenerative potential, offering significant promise for clinical translation.