Hyun Jung Chung

Interferon‐λ (IFN‐λ), a type III interferon that selectively targets epithelial cells, holds strong potential as an intranasal antiviral due to its ability to suppress respiratory virus replication without inducing systemic inflammation. However, clinical translation of human IFN‐λ3 (hIFN‐λ3) is hindered by limited thermostability, protease susceptibility, and rapid mucosal clearance. In this study, instability‐prone elements in hIFN‐λ3 are eliminated through artificial intelligence (AI)‐based backbone remodeling and targeted surface hydrophobic patch engineering. A protease‐sensitive loop is replaced with a de novo α‐helix, which shields neighboring hydrophobic patches and forms a new hydrophobic core, yielding an engineered variant (hIFN‐λ3‐DE1) with enhanced thermostability (Tm > 90 °C), protease resistance, and preserved antiviral activity and structural integrity even after extended heat stress (two weeks at 50 °C). Further glyco‐engineering introduces an N‐linked glycan at a site distant from receptor‐binding interfaces, improving solubility, production yield, and diffusion through synthetic nasal mucus. Intranasal administration of the resulting variant (G‐hIFN‐λ3‐DE1) enables effective mucosal penetration and provides a more rapid onset of in vivo prophylactic protection against influenza A virus. These findings highlight a robust and versatile strategy that combines AI‐driven structural design with glyco‐engineering to develop scalable, bioavailable, and functionally enhanced nasal biologics for respiratory virus prophylaxis.

Fungal infections pose a serious threat to global human health fueled by the increase in immunosuppressive therapies, medical implants, and transplantation. The emergence of multidrug resistance with limited options of current antifungal drugs are a further constraint. There is thus a clear and unmet need to identify therapeutic targets and develop alternative classes of antifungal agents. Here, we hypothesize that dual targeting of key regulatory genes of fungal cell wall synthesis (FKS1 encoding β−1,3-glucan synthase and CHS3 encoding chitin synthase) can synergistically inhibit fungal growth. Based on iterative designs, we generate a small library of fungal-targeted nanoconstructs, and identify a lead construct (FTNx) that shows preferential accumulation in fungal cells over mammalian cells and leads to prominent antifungal effects in vitro. We further show that FTNx is highly effective in a mouse model of disseminated candidiasis, demonstrating diminished fungal growth and enhanced survival rate. This strategy appears promising as an effective treatment for fungal infections in mammalian hosts.

Drug‐resistant microorganisms cause serious problems in human healthcare, leading to the persistence in infections and poor treatment outcomes from conventional therapy. In this study, a gene‐targeting strategy using microbubble‐controlled nanoparticles is introduced that can effectively eliminate biofilms of methicillin‐resistant Staphylococcus aureus (MRSA) in vivo. Biofilm‐targeting nanoparticles (BTN) capable of delivering oligonucleotides are developed that effectively remove biofilm‐associated bacteria upon controlled delivery with diatom‐based microbubblers (MB). The activity of BTN in silencing key bacterial genes related to MRSA biofilm formation ( icaA ), bacterial growth ( ftsZ ), and antimicrobial resistance ( mecA ), as well as their multi‐targeting ability in vitro is validated. The integration of BTN with MB is next examined, resulting in synergistic effects in biofilm removal and antimicrobial activity in an ex vivo porcine skin model. The therapeutic efficacy is further investigated in vivo in a mouse wound model infected with MRSA biofilm, which showed that MB‐controlled BTN delivery substantially reduced bacterial load and led to the effective elimination of the biofilm. This study underscores the potential of the gene silencing platform with physical enhancement as a promising strategy to combat problems related to biofilms and antibiotic resistance.

Interferon-λ (IFN-λ), a type III interferon that selectively targets epithelial cells, holds strong potential as an intranasal antiviral due to its ability to suppress respiratory virus replication without inducing systemic inflammation. However, clinical translation of human IFN-λ3 (hIFN-λ3) is hindered by limited thermostability, protease susceptibility, and rapid mucosal clearance. In this study, instability-prone elements in hIFN-λ3 are eliminated through artificial intelligence (AI)-based backbone remodeling and targeted surface hydrophobic patch engineering. A protease-sensitive loop is replaced with a de novo α-helix, which shields neighboring hydrophobic patches and forms a new hydrophobic core, yielding an engineered variant (hIFN-λ3-DE1) with enhanced thermostability (Tm >90 °C), protease resistance, and preserved antiviral activity and structural integrity even after extended heat stress (two weeks at 50°C). Further glyco-engineering introduces an N-linked glycan at a site distant from receptor-binding interfaces, improving solubility, production yield, and diffusion through synthetic nasal mucus. Intranasal administration of the resulting variant (G-hIFN-λ3-DE1) enables effective mucosal penetration and provides robust prophylactic protection against influenza A virus infection in an in vivo mouse model. These findings highlight a robust and versatile strategy that combines AI-driven structural design with glyco-engineering to develop scalable, bioavailable, and functionally enhanced nasal biologics for respiratory virus prophylaxis.

MRNA therapeutics present a promising strategy for the treatment of human diseases, which requires a carrier to protect the mRNA and ensure its effective cellular delivery. However, research on how the surface physicochemical properties of the mRNA carriers affect the efficiency of delivery of mRNA into cells has been limited thus far. Here, we report that chirality control of lipid nanoparticles (LNPs) can enhance the delivery and transfection efficiency of cargo mRNA into cells. We prepared chiral LNPs by surface modification of LNPs with chiral cobalt oxide nanoparticles. The chiral LNPs with d- or l-chirality resulted in different cellular uptake and transfection efficiencies of luciferase or EGFP mRNA. Chiral LNPs with d-chirality showed significant enhancement in cellular delivery as well as transfection (5.3-fold) of mRNA compared to the control LNPs. This study suggests a novel strategy of using chirality for the delivery of nucleic acid drugs and provides a more in-depth understanding of the effect of chirality on cells.

Respiratory virus infections continue to pose a substantial global health challenge, requiring effective prophylactic and therapeutic strategies. Type III interferon (IFN-λ) has shown promise as an antiviral agent that strongly inhibits viral replication while minimizing systemic inflammation. Intranasal administration of IFN-λ allows easy access to the respiratory mucosa, enhancing localized antiviral responses. However, clinical application of IFN-λ is hindered by rapid mucociliary clearance, limited mucosal adhesion, and susceptibility to proteolytic degradation. Here, we develop nanoliposomes that can deliver IFN-λ through an intranasal route (NLp@IFN-λ) and act as an effective antiviral. We demonstrate that the nanoliposomes enable efficient penetration of IFN-λ in a mucus-mimicking model while allowing controlled release of the protein in vitro. NLp@IFN-λ treatment could effectively up-regulate interferon-stimulated genes in A549 cells, without inducing cytotoxicity. Finally, in vivo delivery of NLp@IFN-λ through a nasal route demonstrates prolonged retention and reduces viral load in nasal tissues in an infection model with influenza virus. This study demonstrates the potential of NLp@IFN-λ as an effective nasal delivery platform for prophylaxis of respiratory virus infections.

An effective delivery platform is crucial for the development of mRNA vaccines and therapeutics. Here, a versatile platform utilizing cholesterol‐modified oligonucleotides (L‐oligo) that bind to the mRNA within lipid nanoparticles (LNP), and enables the effective delivery of the mRNA into target cells is introduced. mRNA incorporated into LNPs via linkage with L‐oligo, termed oligonucleotide‐linked LNP (lnLNP), is superior in cellular uptake and transfection efficiency in target cells in vitro and in vivo, compared to the conventional LNP formulations. It is further applied lnLNP as an mRNA vaccine platform for SARS‐CoV‐2, demonstrating robust induction of neutralizing activity as well as polyfunctional SARS‐CoV‐2‐specific T‐cell response in vivo. The current strategy can be versatilely applied to different LNP platforms, for vaccine and therapeutic applications against various diseases, such as infections and cancers.

Nonviral delivery of the CRISPR/Cas9 system provides great benefits for in vivo gene therapy due to the low risk of side effects. However, in vivo gene editing by delivering the Cas9 ribonucleoprotein (RNP) is challenging due to the poor delivery into target tissues and cells. Here, we introduce an effective delivery method for the CRISPR/Cas9 RNPs by finely tuning the formulation of ionizable lipid nanoparticles. The LNPs delivering CRISPR/Cas9 RNPs (CrLNPs) are demonstrated to induce gene editing with high efficiencies in various cancer cell lines in vitro. Furthermore, we show that CrLNPs can be delivered into tumor tissues with high efficiency, as well as induce significant gene editing in vivo. The current study presents an effective platform for nonviral delivery of the CRISPR/Cas9 system that can be applied as an in vivo gene editing therapeutic for treating various diseases such as cancer and genetic disorders.

The CRISPR/Cas system has been introduced as an innovative tool for therapy, however achieving specific delivery to the target has been a major challenge. Here, an antibody‐CRISPR/Cas conjugate platform that enables specific delivery and target gene editing in HER2‐positive cancer is introduced. The CRISPR/Cas system by replacing specific residues of Cas9 with an unnatural amino acid is engineered, that can be complexed with a nanocarrier and bioorthogonally functionalized with a monoclonal antibody targeting HER2. The resultant antibody‐conjugated CRISPR/Cas nanocomplexes can be specifically delivered and induce gene editing in HER2‐positive cancer cells in vitro. It is demonstrated that the in vivo delivery of the antibody‐CRISPR/Cas nanocomplexes can effectively disrupt the plk1 gene in HER2‐positive ovarian cancer, resulting in substantial suppression of tumor growth. The current study presents a useful therapeutic platform for antibody‐mediated delivery of CRISPR/Cas for the treatment of various cancers and genetic diseases.

Rapid and accurate diagnosis is crucial for managing the global health threat posed by multidrug-resistant bacterial infections; however, current methods have limitations in either being time-consuming, labor-intensive, or requiring instruments with high costs. Addressing these challenges, we introduce a wireless electrochemical sensor integrating the CRISPR/Cas system with electroconductive polymer dot (PD) nanoparticles to rapidly detect bacterial pathogens from human sputum. To enhance the electroconductive properties, we synthesized copper-ion-immobilized PD (PD-Cu), followed by conjugation of the deactivated Cas9 protein (dCas9) onto PD-Cu-coated Si electrodes to generate the dCas9-PD-Cu sensor. The dCas9-PD-Cu sensor integrated with isothermal amplification can specifically detect target nucleic acids of multidrug-resistant bacteria, such as the antibiotic resistance genes kpc-2 and mecA. The dCas9-PD-Cu sensor exhibits high sensitivity, allowing for the detection of ∼54 femtograms of target nucleic acids, based on measuring the changes in resistivity of the Si electrodes through target capture by dCas9. Furthermore, a wireless sensing platform of the dCas9-PD-Cu sensor was established using a Bluetooth module and a microcontroller unit for detection using a smartphone. We demonstrate the feasibility of the platform in diagnosing multidrug-resistant bacterial pneumonia in patients' sputum samples, achieving 92% accuracy. The current study presents a versatile biosensor platform that can overcome the limitations of conventional diagnostics in the clinic.