Biology subjects

Sluijter, J.

Publications and source records attributed to Sluijter, J..

3 recordsLinked to original sources

bioRxiv2026

An extracellular vesicle biogenesis-inspired engineering platform for efficient protein delivery and therapeutic base editing

Efficient and controllable delivery of genome-editing proteins remains a central challenge for therapeutic translation of gene-editing technologies. Extracellular vesicles (EVs) offer an attractive non-viral delivery modality due to their biocompatibility and large capacity for cytosolic cargo delivery. Yet, rational strategies to achieve controlled and programmable protein loading are still lacking. Here, we present NEO-TOP-EVs, an EV biogenesis-guided engineering platform that systematically integrates key features of three design principles inspired by vesicle formation: 1) PI(4,5)P2-mediated plasma membrane targeting, 2) ESCRT-dependent membrane scission, and 3) self-assembly-driven cargo clustering for enabling efficient encapsulation of genome-editing ribonucleoproteins. Together, the NEO design increased cargo incorporation and enhanced functional delivery of gene editing modalities under particle-normalized conditions. Using NEO-TOP-EVs, we achieve efficient delivery of Cas9 and adenine base editor ribonucleoproteins without nucleic acid templates. In an in vitro proof-of-concept, delivery of an adenine base editor targeting proprotein convertase subtilisin/kexin type 9 (PCSK9) induces efficient splice-site disruption, resulting in reduced PCSK9 expression and enhanced LDL receptor activity. Proof-of-concept in vivo experiments provide preliminary evidence of functional Cre protein delivery to the liver. Together, these findings establish NEO-TOP-EVs as a modular platform for protein-based genome editing, demonstrating how biogenesis-informed EV engineering yields functional protein delivery at levels relevant to therapeutic development.

bioengineering↗

bioRxiv2026

Chiral Histidine-Modified Gold Nanoclusters Loaded into Cationic Lipid Nanoparticles for Treatment of Biofilm-associated Infections

Antimicrobial resistance has heightened the risk of device-associated infections, in which Staphylococcus aureus (S. aureus) biofilms persist through extracellular-matrix protection and marked physiological heterogeneity. Gold nanoclusters (AuNCs) can disrupt biofilms via redox-associated stress, but limited in vivo stability and poor local retention constrain their translational potential. Here, we synthesized chiral histidine-modified AuNCs (D-, L-, and DL-AuNCs) and formulated them into mildly cationic lipid nanoparticles (AuNCs@LNP) by microfluidic assembly to enhance biofilm engagement while preserving nanocluster identity. The resulting particles were [~]100-120 nm with high gold loading and a modest charge reversal. In established S. aureus USA300 biofilms, free DL-AuNCs reduced viable burden by up to 5.6 log10 CFU/mL, suppressed metabolic activity, and increased ROS signaling. Notably, LNP encapsulation maintained bactericidal activity and further improved killing by [~]1 log10 CFU at matched Au dose, while substantially enhancing biomass removal (crystal violet residual biomass: 63.77% vs 45.17%), consistent with carrier-mediated matrix destabilization. In a subcutaneous implant infection model, a single perilesional dose of DL-AuNCs@LNP reduced implant-associated bacterial burden by 2.6 log10 CFU per implant versus PBS. These results establish a modular antibiofilm platform that couples nanocluster-driven killing with lipid-facilitated biofilm disruption and defines an efficacy-tolerability window to guide optimization of locally delivered antibiofilm nanomedicines. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=61 SRC="FIGDIR/small/707929v2_ufig1.gif" ALT="Figure 1"> View larger version (22K): org.highwire.dtl.DTLVardef@119134forg.highwire.dtl.DTLVardef@142e7b2org.highwire.dtl.DTLVardef@1799550org.highwire.dtl.DTLVardef@139c33a_HPS_FORMAT_FIGEXP M_FIG C_FIG

microbiology↗

bioRxiv2020

Transcriptional regulation profiling reveals disrupted lipid metabolism in failing hearts with a pathogenic phospholamban mutation

BackgroundThe R14del mutation in the phospholamban (PLN) gene is associated with various types of cardiomyopathies and increases the risk of developing life-threatening ventricular arrhythmias. In this study, we focused on a homogeneous Dutch founder cohort of genetic cardiomyopathy due to PLN R14del mutation and aimed to study the influence of epigenetic changes from a multi-dimensional perspective. ResultsUsing cardiac tissue of PLN R14del patients and donors, we identified differentially acetylated promoters and enhancers (H3K27ac ChIPseq), annotated enriched transcription factor (TF) binding motifs located in those regions, and identified differentially expressed genes (RNA-seq). In line with the fibrofatty replacement in PLN R14del hearts at the histological level, our integrative analysis detected the downregulation of key TF regulators in fatty acid oxidation (FAO) metabolisms and their downstream target in PLN R14del hearts as compared to controls. We further examined heart tissue using immunofluorescence staining (IF) and to confirm the mitochondrial lipid abnormalities in the PLN R14del hearts. Furthermore, we observed the accumulation and deformation of lipid droplets and a disrupted morphology of mitochondria, the key organelle where FAO takes place, in PLN R14del heart using transmission electron microscopy (TEM). ConclusionUsing multi-omics approaches, we successfully obtained a unique list of chromatin regions and genes, including TF-coding genes, which played important roles in the metabolism-related signalling in PLN R14del hearts.

genomics↗