This delivery platform enables the shuttling of engineered proteins throughout all nuclei in multinucleated cells with direct application to gene therapy. Gene therapy has emerged as one of the most promising frontiers in modern medicine, offering potential to treat, cure, or even prevent diseases at their genetic roots. According to industry reports, the global gene therapy market is projected to surpass $20 billion by 2028, with applications ranging from rare genetic disorders and cancers to inherited muscular and neurological diseases. Treating diseases that affect multinucleated cells, such as skeletal muscle fibers, faces significant technical hurdles as these cells contain multiple nuclei. In conventional gene delivery systems, proteins introduced to these cells often localize to only a subset of nuclei, leaving many untreated and reducing the effectiveness of therapies. Additionally, for diseases like muscular dystrophy, where uniform nuclear targeting is essential for restoring muscle function, this limits clinical benefit. For gene therapy to be successful, it must localize to the appropriate cells and, in some cases, subcellular localizations.
Various techniques to achieve this subcellular targeting to the nucleus have generated mixed results. Inclusion of nuclear localization signals (NLS) and nuclear export signals (NES) is effective in facilitating nuclear entry, but in multinucleated cells, such inclusion can inhibit and prevent the effective and efficient spread of such gene therapies.
Researchers at the University of Florida have developed a delivery platform that enables uniform distribution of therapeutic fusion proteins in multinucleated cells. Delivery of nucleic acids encoding multiple cellular migration signals enables the resulting fusion proteins to shuttle out of one nucleus and into adjacent nuclei within the multinucleated cells. This technology provides a platform for the next generation of gene therapy for complex cell types that have historically been difficult to treat with this type of therapy.
This system enables delivery of gene therapy cargos of expression fusion proteins to all nuclei within multinucleated cells, enhancing treatment efficacy
This method of engineering proteins enables efficient, uniform delivery of therapeutic and gene editing proteins with multiple nuclear trafficking signals to target all nuclei in multinucleated cells. The nuclear export signals (NES), nuclear localization signals (NLS), and nucleolar localization signals (NoLS) shuttle these engineered proteins efficiently between the nucleus and cytoplasm. The migration signals are positioned at the N-or C-terminus, or both, and linked via customizable peptide linkers to optimize trafficking dynamics. Delivery is achieved through isolated nucleic acids encoding the fusion protein, which can be packaged in adeno-associated virus (AAV) vectors with tissue-specific promoters for targeted expression in skeletal muscle or liver. The system’s modular design allows researchers and clinicians to adjust the strength and combination of trafficking signals, ensuring balanced nuclear import and export for optimal spread across all nuclei in a multinucleated cell. The ability to shuttle engineered proteins throughout all nuclei in multinucleated cells opens new possibilities for treating genetic diseases that previously had limited options. It means therapies can be more effective and reach every part of the cell that needs help, which is especially important for tissues like muscle that rely on coordinated activity.