Nanobody‐Decorated Lipid Nanoparticles for Enhanced mRNA Delivery to Tumors In Vivo
Pol Escudé Martinez de Castilla, Vincenzo Verdi, Willemijn S. de Voogt, Mariona Estapé Sentí, Arnold C. Koekman, J. Rietveld, Sven van Kempen, Qiangbing Yang, Juliette van Merris, Guido Jenster, Martin E. van Royen, Marcel H.A.M. Fens, Sander A. A. Kooijmans, Wytske M. van Weerden, Guillaume van Niel, Pieter Vader, Raymond M. Schiffelers
Abstract
Prostate cancer (PCa) ranks as the fifth leading cause of cancer-related deaths among men worldwide. In 10-20% of the cases, PCa progresses to an incurable, castration-resistant stage. Castration-resistant PCa cells often overexpress prostate-specific membrane antigen (PSMA), a membrane protein that may serve as their Achilles' heel. Over the past decades, RNA-based therapeutics have emerged as promising treatments for a vast array of diseases, including cancer. In this study, with the ultimate goal of developing a targeted therapy for PCa, lipid nanoparticles (LNPs) are decorated with an anti-PSMA nanobody using click chemistry with a PEG-lipid. Direct stochastic optical reconstruction microscopy (dSTORM) and cluster analysis confirm the presence of at least one nanobody on the surface of 80% of LNPs. These anti-PSMA LNPs exhibit enhanced and specific uptake, and mRNA transfection in PSMA+ cancer cells both in vitro and in a Zebrafish (ZF) metastatic PCa xenograft model. Additionally, in a mouse PSMA-positive xenograft model, systemic administration results in increased LNP accumulation, but not functional mRNA delivery. These findings underscore both the potential and the challenges of using a PSMA-targeted lipid nanoparticle system for mRNA delivery into advanced prostate cancer tumors.