Advancing targeted delivery of intracellular protein therapeutics

About This Grant

PROJECT SUMMARY Intracellular protein delivery has immense potential for research and therapeutic applications, from replacing dysfunctional proteins to modulating key signaling pathways and safely delivering genome editing modalities. However, efficiently delivering functional proteins into the cell cytosol remains a significant challenge. Virus-like particles (VLPs), particularly those derived from retroviruses, offer a promising solution due to their ability to encapsulate large therapeutic proteins and directly deliver them into cells. These cell-derived nanoparticles, composed of assemblies of viral structural proteins such as “gag,” mimic natural viral delivery mechanisms while lacking viral genetic material. Researchers have begun leveraging this capability to transport diverse protein cargoes into cells. Despite their promise, the biogenesis and biological performance of VLPs are influenced by complex factors. For instance, packaging large macromolecules, distinct from the viral genomes they naturally encapsulate, can unpredictably alter their assembly properties. Similarly, their interactions with host factors and serum proteins during production impact assembly, stability, and cellular targeting. Moreover, limited understanding of how these factors influence VLP behavior in vivo, including the role of the protein corona, remains a critical barrier to their advancement as a viable delivery platform for research and clinical applications. To address these challenges, my laboratory will leverage our expertise in biomaterials design, nanoparticle- biological interactions, protein engineering, drug delivery, and nanomedicine to establish design principles for next-generation VLPs with enhanced targeting, potency, and bioproduction efficiency. First, we will develop heuristics for the optimal presentation of bioactive macromolecules on VLPs to enhance cell-specific uptake and targeting. Second, we will elucidate the composition of the VLP protein corona and its impact on in vivo targeting. By understanding how protein corona formation during production and systemic circulation influences VLP performance, we aim to refine their design for improved therapeutic outcomes. Third, we will identify genetic modifiers that regulate VLP assembly, cargo packaging, and release to optimize production yield, stability, and scalability for clinical applications. By elucidating the mechanisms underlying VLP assembly, functionalization, and tropism, the outcome of this work will be a set of design rules to advance the development of VLPs as versatile tools for research and therapeutic applications. These design rules will guide the creation of new VLPs capable of interfacing predictably with biological environments, paving the way for their clinical translation as targeted intracellular delivery platforms. Furthermore, our findings will contribute to the broader understanding and design of other cell-derived nanoparticles for drug delivery applications.