Wei Deng

Choroidal neovascularisation (CNV) is a major cause of irreversible vision loss. Conventional anti-VEGF therapies for CNV often require repeated intravitreal injections, imposing substantial treatment burden and cumulative injection-related risk. Genome editing to disrupt VEGFA could provide durable suppression, potentially from a single administration. However, durability depends on permanently disrupting the target in a well-controlled way, as unrestricted VEGFA loss can cause choriocapillaris atrophy and compromise retinal function. This represents a major safety barrier to clinical translation. In this study, we developed lipid-based nanoparticles to release Cas9/sgRNA ribonucleoprotein (RNP) complexes under light illumination, enabling light-controlled editing of the VEGFA gene. The nanoparticles had a mean hydrodynamic diameter of approximately 130 nm with low polydispersity (0.296 ± 0.032) and high sgRNA encapsulation efficiency (90.33 ± 0.15%). In human retinal pigment epithelial cells, light exposure increased the extent of VEGFA disruption relative to the non-illuminated condition, as confirmed by targeted next-generation sequencing, with corresponding reductions in VEGFA mRNA and protein expression. Following a single intravitreal injection in a laser-induced rat CNV model, the nanoparticles combined with light illumination significantly reduced vascular leakage and CNV-associated structural abnormalities compared with both vehicle and non-illuminated controls, and ocular safety assessment revealed no apparent nanoparticle-related structural abnormalities in the retina. This study provides a foundation for spatially controlled VEGFA genome editing as a potential therapeutic strategy for VEGFA -driven macular neovascular disease.

KRAS mutations, including G12D, G12V and G13D, are major oncogenic drivers in colorectal cancer (CRC) and are associated with therapeutic resistance. However, approved KRAS inhibitors remain focused on G12C subtype, leaving these prevalent variants without approved targeted therapies. Here, we developed a lipid nanoparticle (LNP)-based RNA interference (RNAi) strategy to target these three KRAS variants using a single small interfering RNA (siRNA) sequence. The developed siRNA-LNPs reduced KRAS mRNA and protein expression in HCT116 (G13D), SW620 (G12V) and LS180 (G12D) cells, but not in KRAS wild-type HT29 cells, suppressing the cellular proliferation, clonogenic survival and migration. Transcriptomic profiling revealed changes in KRAS associated signalling, including suppression of cell-cycle and DNA repair programmes, providing a rationale for combining siRNA silencing with the fluoropyrimidine chemotherapeutic agent 5-fluorouracil (5-FU). The combination produced synergistic effects across all three KRAS-mutant CRC cell models and enhanced the apoptosis in both monolayer cultures and 3D spheroids. Collectively, these findings provide proof of concept for this combination strategy and support its further investigation as a potential therapeutic approach for KRAS-mutant CRC. Abstract Figure:

Efficient nuclear delivery of CRISPR/Cas ribonucleoprotein (RNP) is essential for maximising gene-editing efficiency, yet achieving this using non-viral vectors remains a significant challenge. In this study we developed a polymer–lipid hybrid nanoparticle modified with the AS1411 aptamer that targets nucleolin for nucleus-targeted CRISPR/Cas9 delivery. These aptamer-modified nanoparticles were thoroughly characterised and shown to accumulate within cell nuclei, as confirmed by confocal imaging. To further quantitatively analyse nuclear localisation of these nanoparticles, an AI segmentation‑assisted approach based on deep convolutional neural networks (CNN) using the U-Net architecture was employed to segment fluorescent nanoparticles and DAPI signals in microscopy images and quantify colocalisation metrics. After confirming nuclear localisation, we evaluated the gene knockout efficiency of the delivery system by targeting the GFP gene in vitro. The aptamer-functionalized nanoparticles significantly enhanced knockout efficiency, reducing the percentage of GFP-positive cells to 30.0%, compared to 40.8% with untargeted nanoparticles. We further applied the system to knockout the Lcn2 gene, where Cas9/sgRNA complex delivered via targeted nanoparticles achieved higher knockout efficiency than non-targeted counterparts, leading to greater inhibition of breast cancer cell proliferation. These results demonstrate that aptamer-mediated nuclear targeting significantly enhances CRISPR/Cas9 gene editing efficacy and represents a promising strategy to improve transfection efficiency in CRISPR-based gene therapy.

Catalytic nanomedicine represents an attractive therapeutic approach for cancer therapy while heavily relying on the catalytic activity and tumor selectivity of the catalyst. Here, a novel catalytic biohybrid ("bacterobot") of gold nanoparticle-coated electroactive bacterium is developed for enhanced catalytic cancer therapy. The electroactive G. sulfurreducens, for the first time used in cancer therapy, turbocharges the multienzyme-like catalytic activities of biosynthetic gold nanoparticles via cascade catalytic reactions to produce abundant reactive oxygen species to induce apoptosis and pyroptosis of tumor cells. Distinctive tumor accumulation and a tumor inhibition rate of 68% were achieved with single-dose intravenous injection of biohybrids in 4T1 tumor-bearing mice. Overall, the bacterobot biohybrid therapy demonstrates a high degree of therapeutic selectivity to tumor tissues and consequent excellent therapeutic efficacy and systemic biosafety, signifying the potential to create a paradigm shift in the way cancers are treated.

Nanomaterials have become promising platforms in the field of drug and gene delivery, offering unique advantages over traditional therapeutic approaches. Their tunable physicochemical properties enable improved pharmacokinetics and therapeutic performance. A wide range of nanocarriers, including lipid‐based, polymer‐based, and hybrid systems, have been rapidly developed and are attracting increasing attention in both preclinical and clinical research. However, despite promising preclinical outcomes, these systems still encounter critical challenges in achieving precise delivery to specific tissues, cells, and intracellular compartments. This review provides a comprehensive assessment of recent advances in the design and application of nanocarriers for targeted delivery, with emphasis on strategies designed for nuclear targeting. In the context of nuclear targeting, it explores passive approaches involving modulation of particle size, morphology, and surface charge, alongside active targeting strategies incorporating nuclear localization signals and other ligands. In addition to highlighting progress, the review examines the limitations associated with delivery efficiency, off‐target effects, and barriers to clinical translation. By addressing both advances and ongoing challenges, this review provides valuable insights into the design and engineering of targeted nanocarriers. These developments are crucial for unlocking the full potential of precision nanomedicine.