Authors:

Felix Ortiz de Montellano

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About Paper:

Cancer remains a leading cause of mortality worldwide, with current therapies often limited by systemic toxicity and poor specificity. Protein-based nanoparticles offer a promising alternative for the safe and effective delivery of protein therapeutics, including gene-editing therapeutics such as CRISPR/Cas9. Here, elastin-like polypeptide (ELP)/Histone-5 (H5) fusion proteins were engineered to self-assemble into complex coacervate core micelle (C3Ms) nanoparticles for the encapsulation and targeted delivery of therapeutic protein cargos. Using a machine learning platform deployed on Columbia University's high-performance computing cluster, cancer-specific protein minibinders were designed and incorporated into these fusion proteins, enabling the active chemical targeting of specific tumor cells. Utilizing genetically supercharged variants of green fluorescent protein (GFP) 61 as a model cargo, C3Ms were formed and characterized via dynamic light scattering (DLS) and transmission electron microscopy (TEM). Stable micelle formation was observed at a positive charge fraction of 0.8, yielding nanoparticles with a mean diameter of 26.6 nm (number-weighted), a z- average hydrodynamic radius of 41.7 nm, and a low polydispersity index (0.143). Rational engineering of the H5 region enhanced micelle stability under physiological salt conditions through increased charge density and the introduction of disulfide crosslinks. The micelles also displayed pH- responsive disassembly, suggesting potential for endosomal escape and programmed disassembly. Cellular uptake experiments in MDA-MB-231 breast cancer cells revealed enhanced internalization of minibinder- functionalized micelles compared to untargeted controls, as well as evidence of endosomal escape. Collectively, these results demonstrate the feasibility of rationally engineered, fully protein-based nanoparticles as modular, tunable, and biocompatible platforms for targeted intracellular protein therapeutic delivery. Ongoing efforts aim to fully characterize the delivery efficiencies of these particles, and extend this system to Cas9 and other therapeutically relevant proteins, establishing a foundation for protein-based engineered nanocarriers in future cancer therapies.

Source:

Columbia / Biology / 2026

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Co-authors:

Felix Ortiz de Montellano