CRISPR represent a groundbreaking genome-editing technology that has revolutionized genetic modification. This innovative tool offers an unparalleled revolution in the future treatment of genetic disorders, neurological diseases, infectious diseases and cancer. Despite the rapid expansion of CRISPR applications, its clinical use in humans is still relatively limited, with only 69 active clinical trials and 6 completed studies reported so far. This review examined current clinical trials and their processes in addressing various diseases via the CRISPR/Cas system. While earlier literatures have focused mainly on delivery methods and materials for CRISPR/Cas9, our review emphasized innovative targeting conditions and approaches for novel and functional therapeutic designs. In addition, we reviewed recent research to increase the efficiency of CRISPR editing in the management of genetic disorders and cancer, while exploring their future challenges and potential. This review provided a unique perspective on the advancement of CRISPR technology. By addressing these aspects, we aim to contribute to ongoing efforts to improve CRISPR-based therapies and expand their clinical applications, ultimately striving to transform the future of medical treatment.

Wound healing is a series of complex biological events that are tightly coordinated by the body. However, this physiological process is compromised in a pathological setting and underlie conditions including diabetic complications, infectious diseases and chronic inflammatory conditions. Clinically, this translates to multiple pathological features collectively exhibited (microbial colonization, chronic inflammation and impaired regenerative processes). Due to these pathological features, modern wound care approaches have changed from simple passive coverage products to advanced dressing systems which incorporate multifunctional therapies. To comprehensively elucidate recent advancements in advanced wound dressings for drug delivery applications, this review systematically examines material-driven drug release strategies through multiple analytical dimensions. The discussion encompasses structurally engineered controlled-release systems featuring bilayer architectures, layer-by-layer assembly techniques and porous matrix designs, as well as intelligent stimulus-responsive mechanisms based on physicochemical properties, including physical condition modulation, swelling/degradation behavior control, dynamic chemical bond engineering and crosslinking network optimization. Through critical analysis of these cutting-edge technologies, this article provides insightful perspectives on their clinical translation potential and future trajectories.

The therapeutic efficacy and safety of mRNA-based drugs in immunological and nonimmunological applications are critically dependent on the translated protein yield, which requires precise modulation of mRNA expression kinetics. Among the factors influencing mRNA translation, immunogenicity and stability are pivotal in determining the longevity of protein production. Current optimization strategies have integrated (1) molecular engineering (e.g., modified nucleotides), (2) advanced delivery systems (e.g., lipid nanoparticles), and (3) adjuvant drug synergy. This review focuses on co-delivered adjuvant drugs and introduces the concept of "mRNA translation boosters" for the first time. mRNA translation boosters are classified as small-molecule compounds and macromolecular agents that improve translational fidelity through mechanisms including blockade of pattern recognition receptors, modulation of inflammatory cascades, facilitation of endosomal escape, and protection against enzymatic degradation. As clinically validated with COVID-19 mRNA vaccines, these boosters have now demonstrated expanded utility in gene editing therapies and protein replacement applications. This review addresses the immunological challenges encountered during mRNA transfection and translation while summarizing existing mRNA translation boosters that optimize protein expression kinetics. By establishing a mechanistic framework for booster selection and employment, this work provides translational guidance for advancing nucleic acid therapeutics towards their maximum clinical potential.

Lipid nanoparticles (LNPs) have emerged as versatile carriers for the delivery of genetic medicines and small-molecule drugs, offering desired benefits for therapeutic applications. Optimization of the treatment efficacy of nanocarriers necessitates a thorough understanding of the connection between pharmacokinetics and physicochemical properties. This review consolidates scientific efforts to elucidate how LNP's physicochemical attributes influence their in vivo fate, emphasizing particle size and shape, surface electric potential and ligand-binding chemistry. By examining the interplay between LNPs and biological barriers across various administration routes, this review provides insights into tailoring LNP properties for optimal delivery and reduced off-target effects. Recommendations for future research are provided to advance the study of LNP in vivo behaviors and offer a practical framework for optimizing in vivo performance through product design parameters.

Intestinal drug-resistant pathogens, e.g., Salmonella enterica subsp. enterica serovar Typhimurium (S. Tm) and enteropathogenic Escherichia coli (E. coli), frequently cause lifethreatening infectious enteritis. Probiotic-based therapy is a promising way to eliminate drug-resistant pathogens for treatment of infectious enteritis, but its colonizing and therapeutic efficacy after oral administration are limited. Here, we developed a facile therapeutic agent to treat infectious enteritis by co-assembly of the peptide nanodrug melittin-loaded MSN grafted by polysaccharide-binding protein (MMPB) with the famous probiotic bacteria Lactobacillus plantarum (Lac) and Bifidobacterium animalis subsp. lactis (Bif). The nanodrug was composed of the antimicrobial peptide melittin and mesoporous silica nanoparticles exposing the artificial polysaccharide-binding protein. Owing to presence of the artificial protein on the MMPB surface, the nanodrug strongly bound and cross-linked the probiotic cells, forming the Lac + Bif + MMPB co-assembly. During co-incubation with the kanamycin-resistant E. coli strain (Ecka), the co-assembly strongly reduced the viability of Ecka, leading to the increase in the ratio of probiotic to Ecka from 1.6 to 9.2. After oral administration of the co-assembly to the mice pre-colonized by Ecka, Lac + Bif + MMPB almost eliminated the kanamycin-resistant gene in the intestine, and led to 2-3-fold higher levels of the probiotic cells than the nanodrug MMPB or the combined probiotics Lac+Bif. More importantly, in the mice suffering from enteritis caused by drug-resistant S.Tm, the coassembly remarkably recovered the mouse body weight, reduced intestine colonization of S. Tm cells, and decreased the levels of pro-inflammatory cytokines in both serum and colons. This study realized the synthetic biology technique-mediated abiotic/biotic co-assembly for efficient treating infectious enteritis induced by drug-resistant pathogens.

Given the critical shortage of antibody-drug conjugates (ADCs) for bladder cancer (BCa), we developed a novel FGFR3-targeted ADC, LZU-WZLYFG001, composed of a humanized anti-FGFR3 IgG1 monoclonal antibody, a cleavable GGFG linker, and the payload DXD. The antibody was engineered in 293 cells and conjugated via thiol-based chemistry, achieving a drug-to-antibody ratio (DAR) of eight. Comprehensive preclinical assessments, including in vitro and in vivo studies using BCa cells, organoids, cellderived xenograft and patient-derived xenograft (PDX) models, were conducted to evaluate efficacy, targeting ability, mechanism, safety and tissue distribution. LZU-WZLYFG001 demonstrated high purity, targeting specificity and low endotoxin levels, and it significantly inhibited BCa cell proliferation, migration and invasion at nanomolar concentrations, with efficacy strongly associated with FGFR3 expression levels. Mechanistic studies showed binding to FGFR3, internalization and lysosomal release of LZU-WZLYFG001. In organoid and xenograft models, LZU-WZLYFG001 exhibited superior efficacy compared with the gemcitabine + cisplatin (GC) regimen, particularly in GC-resistant PDX tumors, while also showing robust 3D penetration, a bystander effect, and no significant short-term toxicity. Collectively, these findings demonstrate that LZU-WZLYFG001 exhibits excellent preclinical efficacy and safety, and its superiority over GC, together with its activity in resistant tumors, highlights its potential as a novel therapeutic option for BCa.

Local precise drug delivery is conducive to improving therapeutic efficacy and minimizing off-target toxicity. Current local delivery approaches are focused mostly on superficial or postoperative tumor lesions, due to the challenges posed by the inaccessibility of deep-seated tumors. Herein, we report a magnetic continuum soft robot capable of non-invasive and site-specific delivery of prodrug nanoassemblies-loaded hydrogel. The nanoassemblies are co-assembled from redox-responsive docetaxel prodrug and oxaliplatin prodrug, and subsequently embedded into a hydrogel matrix. The hydrogel precursor and crosslinker are synchronously delivered using the soft robot under magnetic guidance and in situ crosslinked at the gastric cancer lesions, forming a drug depot for sustained release and long-lasting treatment. As the hydrogel gradually degrades, the nanoassemblies are internalized by tumor cells. The redox response ability enables them to be selectively activated within tumor cells to trigger the release of docetaxel and oxaliplatin, exerting a synergistic anti-tumor effect. We find that the combination effectively induces immunogenic cell death of gastric tumor, enhancing antitumor immune responses. This strategy offers an intelligent and controllable integration platform for precise drug delivery and combined chemo-immunotherapy.

Integrating photodynamic therapy (PDT) with immunosuppression reversal represents a promising synergistic approach to boost cancer immunotherapy. However, the complicated components and cumbersome preparation procedures of the currently developed nano drug delivery systems heavily hinder their further clinical translation. Herein, a reactive oxygen species (ROS)/photo dual-responsive amphipathic prodrug (denoted as PPTN) was designed and synthesized by linking NLG919, an indoleamine-2,3-dioxygenase (IDO) inhibitor, with the photosensitizer protoporphyrin IX (PpIX) by a thioketal moiety, and further modifying with $\left(\mathrm{82373811,82574333,82504683}\right)$. PPTN could self-assemble into nanoscale unimolecular micelles in aqueous solution without additional excipients, increasing tumor accumulation while effectively addressing the pronounced hydrophobicity challenge of PpIX. Upon light exposure, PPTN generated ROS, not only directly damaging cancer cells, but also trigger the breakage of thioketal bond to accelerate simultaneous release of NLG919. Therefore, PPTN potentially act as a promising ROS/photo dual-responsive carrier-free prodrug delivery system for controllable drug release and specific tumor therapy. Moreover, PPTN induced simultaneous PDT-triggered immunogenic cell death (ICD) effect and specific IDO blockade to boost immune response, exhibiting potent suppression efficacy against primary and distant tumors. Overall, with the superiorities of easily controllable preparation procedures, synchronous drug delivery and ROS/photo dual-responsiveness, such a prodrug unimolecular micelle may represent a promising nanoplatform for photoactivated-immunotherapy.

Metabolic dysfunction-associated fatty liver disease (MASLD) and alcohol-associated liver disease (ALD) are prevalent chronic liver diseases that can progress to steatohepatitis, fibrosis, cirrhosis, and ultimately liver failure. Here, we demonstrated that oral administration of GNVs provided substantial protection against liver injury and fibrosis in MASLD and ALD mouse models. In a Western-style high-fat diet-induced MASLD model and a chronic binge alcohol-induced ALD model, GNVs treatment significantly reduced gut leakiness by restoring intestinal junctional complex proteins and rebalancing the gut microbiome. GNVs attenuated hepatic lipid accumulation, oxidative stress and fibrogenic markers. GNV treatment downregulated the fibrosis-associated tissue inhibitor of metalloproteinase-2 (TIMP2) pathway in hepatic stellate cells, which is linked to enhanced matrix degradation and reduced fibrogenesis. GNVs prevent MASLD- and ALD-associated gut barrier dysfunction and liver fibrosis through modulation of the gut-liver axis and the TIMP2 pathway. Edible GNVs represent a novel, multifaceted therapeutic strategy for managing chronic liver diseases.

This study aimed to assess the therapeutic potential of nisin, 5 -fluorouracil (5FU) and selenium encapsulated in folate-conjugated thiolated chitosan nanoparticles (N/5FU/Se@FTCsNPs), combined with a probiotic cocktail of Lactobacillus acidophilus and Bifidobacterium bifidum, against colorectal cancer (CRC). The nanoparticles ( 277 nm, +9.2 mV ) exhibited high drug loading efficiencies (5FU: 89.11%, nisin: 70.68%) and pHresponsive release, with minimal drug release under gastric conditions and ∼60.7% release at colonic pH, facilitating targeted delivery. The formulation remained stable for over 40 d at $-{20}^{\circ }\mathrm{C}$ and ${4}^{\circ }\mathrm{C}$, demonstrating excellent biocompatibility ( <2% hemolysis) and exhibiting strong mucoadhesive and mucus-penetrating abilities. In vitro, N/5FU/Se@FTCsNPs selectively targeted CT26 colon cancer cells ( ${\mathrm{I}\mathrm{C}}_{5n}:1.57\mu \text{ }\mathrm{g}/\mathrm{m}\mathrm{l}$ ) with minimal effects on healthy cells, enhanced cellular uptake, and induced ROS-mediated apoptosis. In vivo, oral administration-especially with probiotics-significantly reduced tumor volume, improved survival rates and alleviated chemotherapy-related side effects such as diarrhea and weight loss. Biodistribution studies confirmed increased tumor targeting and decreased off-target exposure. Mechanistically, the treatment downregulated oncogenes and inflammatory markers ( 2 - to 12.5 -fold), including β-catenin, mTOR, COX- 2 and VEGF- $\alpha $, while upregulating tumor suppressors and protective genes ( 4 to 14.8 fold), such as PTEN, CASP9 and Mucin 2(P<0.0001). This indicates inhibition of proliferation, metastasis, inflammation, and angiogenesis, along with improved gut barrier function. Cytokine profiling and histological analysis further confirmed reduced systemic inflammation and maintained hematological safety. These findings highlight N/5FU/Se@FTCsNPs combined with probiotics as a promising, safe and effective oral therapy for CRC, leveraging microbiota modulation and targeted delivery.

Multidrug-resistant Klebsiella pneumoniae (MDR-KP) is characterized by high mortality and risk of nosocomial transmission, and biofilm constitutes the primary challenge in the treatment of its implant-associated and refractory pulmonary infections. Notably, the hypoxic microenvironment and the physical barrier of biofilm leading to the increased tolerance of the bacteria to antibiotics. Herein, a hypoxia-responsive hybrid nanoparticle (CHLip@FLD/COL) loaded separately with anti-biofilm candidate fingolimod (FLD) and antibiotic colistin (COL) is achieved targeting antibacterial efficacy against MDR-KP in vitro and in vivo. CHLip@FLD/COL is composed of hybridizing hypoxia-responsive lipids (HLipid) and lipid A targeting materials DSPE-mPEG-COL. HLipid is synthesized by hexadecanedioic acid esterified with nitroimidazole, while DSPE-mPEG is coupling with vector COL via amide reaction. The relative level of extracellular polymeric substances and the NIR-IIb sO₂ images of the infection site are used as indicators to establish mature biofilm models. CHLip@FLD/COL readily releases FLD and COL in hypoxic conditions, and its MIC against MDR-KP is only one-sixteenth of that when COL is used alone in vitro. The nanoparticle exhibits bacterial targeting ability and antibacterial effect in the pulmonary infection and biofilm infection mice models. Bacterial loads eliminated by $4{\mathrm{l}\mathrm{o}\mathrm{g}}_{10}$ CFU and $2{\mathrm{l}\mathrm{o}\mathrm{g}}_{10}$ CFU, respectively. The strategy provides a valuable reference for the treatment of refractory infections caused by MDR-KP.