Gaeun Kim

Small extracellular vesicles (sEVs) have emerged as next‐generation multifunctional nanotherapeutics due to their parental‐cell traits and role in intercellular communication. Among them, immune cell–derived sEVs are uniquely positioned to couple innate immunomodulatory activities with therapeutic payload delivery, making them highly attractive for cancer therapy. In particular, M1 macrophage‐derived sEVs (M1‐sEVs) preserve the tumour‐suppressive functions of their parent cells, including tumour microenvironment (TME) reprogramming, immune activation, and inhibition of cancer progression. However, the mechanisms by which these activities are coordinated within the TME, and whether they act independently or synergistically, remain poorly understood. Clarifying these mechanisms is crucial for harnessing their intrinsic bioactivity in combination with their natural capacity as drug delivery nanocarriers to optimize therapeutic efficacy. Here, we demonstrate that M1‐sEVs exhibit intrinsic stability and circulation longevity via ‘do not eat me’ ligands, as well as tumour‐homing ability revealed by proteomic profiling, enabling efficient uptake and deep infiltration in breast cancer models. Functionally, M1‐sEVs deliver antiproliferative microRNAs that suppress tumour metabolism, growth, and progression by inhibiting self‐renewal, adhesion, migration, motility, and invasion. Importantly, by integrating this endogenous bioactivity with exogenous doxorubicin loading, we achieved synergistic efficacy: a 3‐fold reduction in IC ₅₀ in vitro (0.46 µM vs. 1.45 µM for free drug) and 70.18% tumour growth inhibition in vivo. These findings highlight M1‐sEVs as dual‐action nanotherapeutics that combine innate immune‐regulatory and tumour‐inhibitory functions with efficient drug delivery, advancing their development as powerful platforms for cancer therapy.

Small extracellular vesicles (sEVs) have emerged as next-generation multifunctional nanotherapeutics due to their parental-cell traits and role in intercellular communication. Among them, immune cell–derived sEVs are uniquely positioned to couple innate immunomodulatory activities with therapeutic payload delivery, making them highly attractive for cancer therapy. In particular, M1 macrophage-derived sEVs (M1-sEVs) preserve the tumor-suppressive functions of their parent cells, including tumor microenvironment (TME) reprogramming, immune activation, and inhibition of cancer progression. However, the mechanisms by which these activities are coordinated within the TME, and whether they act independently or synergistically, remain poorly understood. Clarifying these mechanisms is crucial for harnessing their intrinsic bioactivity in combination with their natural capacity as drug delivery nanocarriers to optimize therapeutic efficacy. Here, we demonstrate that M1-sEVs exhibit intrinsic stability and circulation longevity via ‘do not eat me’ ligands, as well as tumor-homing ability revealed by proteomic profiling, enabling efficient uptake and deep infiltration in breast cancer models. Functionally, M1-sEVs deliver antiproliferative microRNAs that suppress tumor metabolism, growth, and progression by inhibiting self-renewal, adhesion, migration, motility, and invasion. Importantly, by integrating this endogenous bioactivity with exogenous doxorubicin loading, we achieved synergistic efficacy: a 3-fold reduction in IC ₅₀ in vitro (0.46 μM vs. 1.45 μM for free drug) and 70.18% tumor growth inhibition in vivo . These findings highlight M1-sEVs as dual-action nanotherapeutics that combine innate immune-regulatory and tumor-inhibitory functions with efficient drug delivery, advancing their development as powerful platforms for cancer therapy.

High-content, high-throughput (HCHT) screening platforms are essential for drug discovery, yet conventional 2D assays lack physiological relevance, and current 3D spheroid systems often face challenges to scalability, uniformity, and the analytical efficiency required for statistically robust screening. Here, we present a fully integrated 3D HCHT platform that synergizes tumor spheroid arrays generated from a bioinert inverted colloidal crystal (iCC) hydrogel framework with an automated, high-speed image analysis pipeline for rapid and spatially resolved therapeutic profiling. The iCC framework enables spontaneous self-assembly of highly ordered tumor spheroid array at high spheroid density (∼79.8 spheroids·mm^-2) with tight size uniformity (-1). The automated image-processing algorithm achieves robust region-of-interest segmentation, fluorescence-weighted-spheroid centroiding (FwSC), and multiparametric spatial analysis in <5 s per image─markedly faster than well-plate analysis (e.g., ∼5 min per 96-well plate) or sequential single-spheroid analysis methods (∼1 s per spheroid). Using this platform, we capture dose-dependent diffusion of doxorubicin, tumor penetration profile of small extracellular vesicles, and cell-type-specific infiltration behaviors of monocytes versus macrophages. Comparative viability profiling across chemotherapeutics further reveals distinct spatial toxicity signatures, highlighting the importance of spatial context in drug response assessment. Collectively, this platform enables rapid, reproducible, and spatially informative screening in 3D, offering a powerful tool for drug discovery, tumor modeling, and immunotherapeutic development.

Tauopathies, a group of neurodegenerative disorders, are characterized by the abnormal aggregation of tau proteins into neurofibrillary tangles, driving synaptic dysfunction, neuronal loss, and disease progression through tau aggregate propagation. Graphene quantum dots (GQDs) functionalized with D ‐cysteine ( D ‐GQDs) have shown promise in inhibiting tau aggregation and transmission via π–π stacking and electrostatic interactions with tau proteins. However, the nonspecific binding of GQDs to various proteins in the physiological environment, such as serum albumin, limits their clinical translation. In this study, the aim is to enhance the specificity of D ‐GQDs toward tau protein by incorporating a tau‐targeting N‐amino peptide, mxyl‐NAP2. The mxyl‐NAP2/ D ‐GQD complex demonstrates improved selectivity for tau protein over serum albumin, effectively enhancing the inhibition of tau aggregation. To further minimize off‐target effects and optimize therapeutic delivery, the mxyl‐NAP2/ D ‐GQD complex is loaded into small extracellular vesicles (sEVs), followed by functionalization of sEVs with neuron‐targeting ligand, rabies viral glycoprotein peptides. This strategy not only reduces off‐target effects, but also enhances uptake by neuron cells, which further improves inhibition of tau transmission between neurons. The results indicate that mxyl‐NAP2/ D ‐GQD‐sEVs hold great promise for overcoming the off‐target limitations of D ‐GQDs and advancing the development of precision therapeutics for neurodegenerative diseases.

High-content, high-throughput (HCHT) screening platforms are essential for drug discovery, yet conventional 2D assays lack physiological relevance, and current 3D spheroid systems often face challenges to scalability, uniformity, and the analytical efficiency required for statistically robust screening. Here, we present a fully integrated 3D HCHT platform that synergizes tumor spheroid arrays generated from a bioinert inverted colloidal crystal (iCC) hydrogel frame-work with an automated, high-speed image analysis pipeline for rapid and spatially resolved therapeutic profiling. The iCC framework enables spontaneous self-assembly of highly-ordered tumor spheroid array at high spheroid density (∼79.8 spheroids·mm ^{− 2} ) with tight size uniformity ( − 1 ). The automated image-processing algorithm achieves robust region-of-interest segmentation, fluorescence-weighted centroiding, and multi-parametric spatial analysis in versus macrophages. Comparative viability profiling across chemotherapeutics further reveals distinct spatial toxicity signatures, highlighting the importance of spatial context in drug response assessment. Collectively, this platform enables rapid, reproducible, and spatially informative screening in 3D, offering a powerful tool for drug discovery, tumor modeling, and immunotherapeutic development.

Small extracellular vesicles (sEVs) are promising nanocarriers for drug delivery to treat a wide range of diseases due to their natural origin and innate homing properties. However, suboptimal therapeutic effects, attributed to ineffective targeting, limited lysosomal escape, and insufficient delivery, remain challenges in effectively delivering therapeutic cargo. Despite advances in sEV-based drug delivery systems, conventional approaches need improvement to address low drug-loading efficiency and to develop surface functionalization techniques for precise targeting of cells of interest, all while preserving the membrane integrity of sEVs. We report an enhanced gene delivery system using multifunctional sEVs for highly efficient siRNA loading and delivery. The integration of chiral graphene quantum dots enhanced the loading capacity while preserving the structural integrity of the sEVs. Additionally, lysosomal escape is facilitated by functionalizing sEVs with pH-responsive peptides, fully harnessing the inherent homing effect of sEVs for targeted and precise delivery. These sEVs achieved a 1.74-fold increase in cytosolic cargo delivery compared to unmodified sEVs, resulting in substantial gene silencing of around 73%. Our approach has significant potential to advance sEV-based gene delivery in order to accelerate clinical progress.