Search Results (263)

Lipid nanoparticles (LNPs) have become an important platform for the delivery of RNA therapeutics, including messenger RNA (mRNA) and small interfering RNA (siRNA). However, most clinically approved LNP formulations exhibit strong liver tropism following systemic administration, which limits efficient delivery to extrahepatic tissues. This inherent biodistribution profile has therefore been recognized as a key challenge for expanding the therapeutic applications of RNA nanomedicine. Recent efforts have focused on engineering functionalized LNP systems to improve delivery specificity beyond the liver. Surface modification with targeting ligands—such as antibodies, peptides, and nucleic acid aptamers—can promote receptor-mediated uptake by specific immune cell populations, including macrophages, dendritic cells and T lymphocytes. In parallel, advances in lipid design have improved intracellular RNA delivery by facilitating endosomal escape. These developments have broadened the potential use of RNA nanomedicine for inflammatory disorders, including autoimmune diseases, neuroinflammation, and cardiovascular inflammation. Functionalized LNPs are also being investigated for in vivo engineering of immune cells. This review summarizes current strategies for designing functionalized LNP systems, highlights their emerging applications in immune and inflammatory diseases, and discusses key challenges for clinical translation. Full article

Background: Nanostructured, rod-shaped microparticles represent a promising drug delivery platform for the pulmonary delivery and targeting of alveolar macrophages by exploiting the aerodynamic advantages of fiber-like geometries. These microrods feature a hierarchical architecture, designed for potential macromolecular payloads, and silica (SiO₂)-based systems have previously been shown to successfully deliver oligonucleotides in vitro. However, current microrod systems mainly rely on nanoparticulate SiO₂-based frameworks with limited biodegradability and lack a specific escape mechanism to the cytosol. Therefore, a nanostructured calcium phosphate (CaP) framework is proposed as a biodegradable and resorbable alternative, featuring pH-responsive dissolution under endolysosomal conditions. Methods and Results: This study presents the fabrication of nanostructured, rod-shaped calcium phosphate microparticles and discusses their suitability as a potential pulmonary drug delivery platform. The particles feature dissolution-driven disintegration in acidic and ion-rich environments relevant to phagolysosomes. In addition, the particles exhibited a favorable acute cytotoxicity profile in the murine alveolar macrophage cell line MH-S compared with their SiO₂-based counterparts. Comparative RNA-seq analysis of MH-S exposed to the particles indicates a mild transcriptomic response, while canonical signatures of classical or alternative macrophage activation programs were not observed, supporting a generally well-tolerated exposure profile of the carrier. Conclusions: Together, these findings establish key prerequisites for employing calcium phosphate microrods as a biodegradable pulmonary carrier platform in subsequent studies incorporating therapeutic cargos. Full article

Nanomaterials are emerging versatile platforms for therapeutic delivery, as they offer precise control over drug, antioxidant, and genetic payload transport across biological barriers. Inorganic, organic, hybrid, and biomimetic systems are the major classes of nanomaterials, which all have different physicochemical properties such as size, surface charge, and surface functionalization. These properties collectively influence stability, biodistribution, cellular uptake, and release kinetics. Engineering strategies are increasingly using stimuli-responsive designs that are triggered by pH, reactive oxygen species (ROS), and intracellular redox gradients to perform spatially and temporally controlled delivery. Antioxidant and redox-modulating nanocarriers are of great importance as they overcome the limited bioavailability and nonspecific activity of conventional antioxidants by improving stability, targeting oxidative microenvironments, and allowing for regulated release. Improvements in lipid, polymeric, and inorganic nanoplatforms have also developed gene delivery applications, including siRNA, mRNA, and CRISPR/Cas systems, to provide better cytosolic release and precise therapeutics. When diagnostic imaging is integrated with therapy through theranostic nanoparticles, real-time monitoring and personalized intervention are possible. Safety, scalable manufacturing, and regulatory alignment are some challenges that show the need for standardization and translational procedures to utilize the potential of theranostic nanomedicine. Full article

The efficient delivery of small interfering RNAs (siRNAs) to immune and respiratory cells represents a key methodological challenge in developing inhaled RNA interference (RNAi) approaches. A central question is whether siRNA functionality is preserved following aerosolization, as the mechanical stress of nebulization may compromise siRNA integrity and silencing activity. Here, we report a proof-of-concept study using THP-1-derived macrophage-like cells as a tractable in vitro model to characterize jet nebulization for siRNA delivery. Three siRNA candidates targeting interleukin-1 beta (IL-1β) were computationally designed and validated for potent silencing activity and low cytotoxicity. Using a commercially available, off-the-shelf jet nebulizer combined with Lipofectamine RNAiMAX, we demonstrate that siRNA-lipoplexes retain their gene-silencing activity after aerosolization, achieving robust IL-1β knockdown. The delivery efficiency was influenced by siRNA-lipoplex complexation, highlighting the importance of formulation parameters. These findings establish a practical and accessible in vitro platform for evaluating nebulized siRNA functionality, providing a foundation for future studies in more complex and physiologically relevant airway models. Full article

Hypocrellin A (HA) represents a pharmaceutically important perylenequinone photosensitizer produced by Shiraia bambusicola. However, submerged fermentation remains constrained by filamentous morphological characteristics and inherent mass transfer limitations. Although microparticle-enhanced cultivation (MPEC) has demonstrated efficacy in filamentous fungal systems, the molecular mechanisms by which physical cues, such as microparticle-induced shear stress, reprogram fungal metabolism remain largely unexplored. This work systematically optimizes SiO₂-based MPEC parameters for S. bambusicola GDMCC 60438, including particle dimensions, temporal addition protocols, and solid loading. Mechanistic investigations integrated pellet morphology analysis, membrane lipid composition, intracellular redox status, energy/precursor markers, and RNA-seq transcriptomic profiling with qRT-PCR validation. Under optimized conditions (10% w/v SiO₂, 30 mesh, added at 6 h), HA yield reached 41.76 ± 5.02 mg/L, representing a 3.65-fold increase over controls. MPEC shifted morphology toward smaller, more porous pellets with denser internal structure, accompanied by increased membrane fluidity (unsaturated/saturated fatty acid ratio from 1.54 to 2.63), elevated ROS levels with antioxidant enzyme activation, and enhanced acetyl-CoA and ATP accumulation. Transcriptomic analysis identified 206 differentially expressed genes enriched in oxidative phosphorylation, carbon metabolism, and stress responses, with upregulation of PKS-related biosynthetic genes and major facilitator superfamily transporters. This work establishes an integrated mechanistic framework linking particle-induced morphological changes to metabolic reprogramming through oxidative stress and subsequent transcriptional activation of the HA biosynthetic pathway, providing rational design principles for MPEC strategies in filamentous fungi. Full article

Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease for which there is currently no cure. Dominant mutations in the TARDBP gene are causative of ALS. In particular, the p. G376D substitution in TDP-43 causes familial ALS and it is associated with TDP-43 mislocalization in the cytosol, increased presence of cytoplasmic aggregates, and lysosomal and mitochondrial dysfunction. We previously designed a small interfering RNA (siRNA) that specifically targets and silences the mutant allele and we demonstrated that, in patient-derived fibroblasts, it can reduce TDP-43 aggregation, decrease oxidative stress, and improve cell viability. Here, we investigated the ability of this siRNA to revert some ALS-associated pathological phenotypes in motor neurons derived from induced pluripotent stem cells (iPSCs), as motor neurons are the primary cells affected in ALS. siRNA treatment reduced TDP-43 mislocalization, enhanced lysosomal function and cell viability, and decreased oxidative stress. These findings indicate that this allele-specific siRNA effectively reverses key ALS-related cellular deficits in motor neurons, representing a promising candidate for targeted therapy in patients carrying the TDP-43 G376D mutation. Full article

Understanding species-dependent siRNA length generation provides both fundamental and applied insights. At the basic level, it highlights an underappreciated dimension of RNAi diversity in insects, emphasizing that antiviral immunity cannot be fully understood from Drosophila melanogaster alone. At the applied level, these findings have direct implications for the design of dsRNA-based pest management strategies, where tailoring siRNA production to the target insect order could enhance both efficacy and specificity. Previously studies demonstrated that Dicer-2-generated siRNAs exhibit distinct, species-dependent length distributions: dipterans (D. melanogaster) and coleopterans (Tribolium castaneum) mainly produce 21-nt siRNAs; and hymenopterans (Bombus terrestris) and orthopterans (Locusta migratoria) generate siRNAs enriched at 22 nt; whereas lepidopterans such as Spodoptera exigua and Trichoplusia ni predominantly produce 20-nt siRNAs. The central biological question motivating this study was whether structural divergence in Dicer-2 may explain these lineage-specific differences in siRNA length profiles and antiviral RNAi efficiency. To address this, we interpreted observed structural variations in the context of the “molecular ruler” hypothesis and integrated them with previous experimental data on siRNA length variation across insect taxa. Future studies that combine high-resolution structural biology, comparative genomics, and functional assays will be essential to experimentally test whether the structural correlations proposed here determine Dicer-2 cleavage length in vivo and in vitro, and to leverage this knowledge for both agricultural and biomedical applications. Full article

Early detection of hepatocellular carcinoma (HCC) remains challenging due to limitations including the lack of reliable biomarkers. While molecular diagnostics hold promise, their use is limited because tissue biopsies are not routinely performed in HCC. Long non-coding RNAs (lncRNA), such as HOXA transcript at the distal tip (HOTTIP), have been implicated in HCC, with single-nucleotide polymorphisms forming haplotypes that may influence disease progression. This study investigated the clinical relevance of HOTTIP SNPs rs17501292 and rs2067087 in 198 Egyptian HCC patients (129 non-metastatic, 69 metastatic). Moreover, molecular docking was used to design small interfering RNAs (siRNAs) targeting HOTTIP. Genotypes TT and TG (rs17501292) and GG and GC (rs2067087) were associated with reduced metastatic risk compared to GG and CC genotypes, respectively. Survival analysis revealed that TT (rs17501292) and GC (rs2067087) genotypes correlated with improved outcomes. ROC curve analysis confirmed the diagnostic and prognostic value of specific genetic models, affirming their value as biomarkers for metastasis and survival. Molecular docking identified two promising therapeutic candidates. Overall, we can conclude that HOTTIP SNPs may serve as promising potential non-invasive biomarkers for HCC metastasis and prognosis, while the identified siRNAs offer a novel targeted therapeutic approach. Full article

Objectives Breast cancer remains a major cause of female cancer-related deaths, with current therapies limited by poor tumor targeting and an immunosuppressive microenvironment. This study designed CA/ZYII-siP-c-L—an asymmetric lipid bilayer-coated calcium/aluminum (CA)-core nanoparticle—to co-deliver PD-L1 siRNA (siP) and ziyuglycoside II (ZYII) to boost therapeutic efficacy. Methods CA/ZYII-siP-c-L was fabricated through modified microemulsification to first construct the CA cores, followed by thin-film hydration for encapsulation of ZYII within the hydrophobic domain, and via hybridization of the outer lipid layer with DSPE-PEG1000-PAMAM to finally enable specific adsorption of siP. The characterization of CA/ZYII-siP-c-L was performed to get size distribution, zeta potential and in vitro release behavior. In vitro cytotoxicity of the nanoparticles to NIH3T3 and 4T1 cells was detected by the CCK-8 method. The uptake capacity to 4T1 breast cancer cells was determined using inductively coupled plasma optical emission spectrometry and high-performance liquid chromatography. Pharmacokinetic studies and tissue distribution experiments were performed. In BALB/c mice bearing orthotopic 4T1 tumors, efficacy evaluations were conducted with the detection of tumor immune microenvironment; meanwhile, organ damage was evaluated by hematoxylin-eosin staining of major organs and detection of routine biochemical indicators. Results CA/ZYII-siP-c-L was characterized by dynamic light scattering (mean size ~185.7 nm) and zeta potential analysis (~9.35 mV). In vitro, the nanoparticle exhibited low cytotoxicity in NIH3T3 normal cells, high uptake by 4T1 breast cancer cells, and pH-responsive release. For the pharmacokinetic study, CA nanoparticle system could significantly enhance the systemic exposure of ZYII, compared to free ZYII suspension. In BALB/c mice with orthotopic 4T1 tumors, CA/ZYII-siP-c-L accumulation in tumors was 3.5-fold higher than that of free drugs, significantly enriching helper T cells and cytotoxic T lymphocytes while reducing regulatory T cells and suppressive dendritic cells in the tumor immune microenvironment; this immunomodulatory effect, combined with PD-L1 silencing at protein levels, contributed to ~62% inhibition of tumor growth with no organ damage (confirmed by hematoxylin and eosin staining of major organs and normal biochemical indices). Conclusions CA/ZYII-siP-c-L integrates safety, targeting, and codelivery capabilities, offering a promising strategy for breast cancer treatment by combining siP-mediated immunity regulation and the antitumor effects of ZYII. Full article

Gene delivery/administration and, in particular, small interfering RNA (siRNA) delivery represent a therapeutic challenge, though very effective carriers have yet to be identified. Cyclodextrins (CDs) are cyclic oligosaccharides with unique host–guest inclusion capabilities, widely recognized in the pharmaceutical field for their ability to enhance drug solubility and bioavailability. Their excellent biocompatibility and chemical versatility make them powerful building blocks for the design of supramolecular nanovectors (NVs). Thanks to their facility of functionalization, CDs are highly versatile and have found numerous applications across various fields. In this context, CD-based NVs are currently explored as non-viral agents to transport and release siRNA. Recent studies suggest that self-assembled NVs based on CDs can improve the transfection and safety of siRNA delivery. This review provides a comprehensive overview of the most recent advances in the design of NVs based on CDs and their use for siRNA delivery, discussing the role played by structural differences and chemical functionalization in the context of encapsulation and release. Full article

Rapid depressurization and warming during recovery can trigger stress in deep-sea microbes and accelerate RNA degradation. We developed a remotely operated vehicle (ROV)-oriented multi-sequence microbial sampler for 2000 m sampling (20 MPa, 2 °C) that integrates in situ filtration with immediate RNAlater injection (an RNA stabilization reagent), collecting up to 12 samples per dive. A Dirichlet sampling–B-spline–SVM framework was used to optimize the cam profile of the sequence trigger for robust actuation under geometric constraints and realistic tolerances in both manufacturing and assembly. Relative to the baseline 3-4-5 motion law, the optimized design reduces nominal peak driving torque by ~18–20% and lowers the maximum torque under tolerance perturbations; tests show a further ~10–25% reduction using a SiC ball–ZrO₂ block pair versus a MoS₂-lubricated titanium pushrod–ZrO₂ block pair. A Darcy–Forchheimer porous-media computational fluid dynamics (CFD) model predicts earlier clogging on the lower membrane and a fast-to-slow RNAlater displacement process; greater membrane resistance mismatch delays 95% displacement and increases RNAlater loss. Simulations and Rhodamine B tests suggest an RNAlater consumption of 0.9 L per parallel filter (one membrane per side), and 20 MPa chamber tests confirm stable operation and membrane retrieval. Full article

The development and application of therapeutic oligonucleotides, such as siRNA, miRNA, ASOs and aptamers, is a rapidly growing field in biomedicine. These molecules are undergoing extensive preclinical and clinical testing, and the market for synthetic RNA drugs is expanding. However, several challenges remain, including targeted delivery and high costs associated with development, screening and production. One significant advance has been the creation of GalNAc-conjugates, which selectively target ASGPR and deliver oligonucleotides to hepatocytes. Although these conjugates have shown promising results, their widespread use is limited by the lack of effective synthesis methods. Thus, the development of new methods for the synthesis of ligand-oligonucleotide conjugates is an important task to which this study is devoted. In this study, we created a library of siRNA conjugates with the GalNAc L-96 ligand to suppress the expression of the PCSK9 gene associated with elevated LDL and an increased risk of developing cardiovascular diseases. The selection of the most effective siRNA molecules was carried out using an algorithm previously developed by our research group, which considers thermodynamic stability, predicted specificity and effectiveness. To experimentally confirm the effectiveness of conjugates, an in vitro model based on the cultivation of hepatocyte cells was developed. Optimization of the conjugate synthesis process has significantly reduced the cost of manufacturing technology, which creates the potential for efficient scaling of synthesis for transfer and application in the pharmaceutical industry. The results of the study showed that the development of the siRNA sequence optimized in silico resulted in a significant increase in the inhibitory effect of the GalNAc-siRNA conjugate compared to a compound similar to a commercial drug. Full article

A 19 nt fragment that spans the SARS-CoV-2 furin cleavage site (FCS) is identical to the reverse complement of a proprietary human DNA repair gene sequence. Rather than interpreting this overlap as evidence of a laboratory event, this article uses it as a theoretical springboard to explore underappreciated biorisk concerns, specifically in the context of cancer research. Although they are RNA viruses, coronaviruses are capable of hijacking host DNA damage response (DDR) pathways, exploiting nuclear functions to enhance replication and evade innate immunity. Under selective pressures (antivirals, DDR antagonists, or large-scale siRNA libraries designed to silence critical host genes), escape mutants may arise with fitness advantages. Parallel observations involving in vivo RNA interference via chimeric viruses lend plausibility to some of the key aspects underlying unappreciated biorisks. The mechanistic insights that incorporate DNA repair mechanisms, CoVs in the nucleus, specifics of viruses in cancer research, anticancer drugs, automated gene silencing experiments, and gene sequence overlaps identify gaps in biorisk policies, even those unaccounted for by the potent “Sequences of Concern” paradigm. Key concerning attributes, including genome multifunctionality, such as NLS/FCS in SARS-CoV-2, antisense sequences, and their combination, are further described in more general terms. The article concludes with recommendations pairing modern technical safeguards with enduring ethical principles. Full article

As a natural flavonoid, the flavonoid luteolin is characterized by its powerful antioxidant and anti-inflammatory effects. While its precise mechanisms require further elucidation, existing evidence confirms its efficacy in ameliorating myocardial ischemia–reperfusion injury (MIRI). This research was designed to investigate the mechanism through which luteolin protects against MIRI. We established MIRI rat models through the ligation of left anterior descending coronary artery (LAD). To evaluate the cardioprotective effects of luteolin, echocardiographic analysis was performed, Hematoxylin and Eosin (HE) staining, and serum cardiac injury markers creatine kinase-MB (CK-MB) and lactate dehydrogenase (LDH). Cardiac vascular permeability was determined using Evans blue staining. To mimic ischemia–reperfusion injury, endothelial cells (ECs) were subjected to oxygen-glucose deprivation/reoxygenation (OGD/R) in vitro. Endothelial cell barrier function was evaluated through F-actin phalloidin staining and FITC-Dextran fluorescence leakage experiments. To elucidate the molecular mechanism, FOXP1 small interfering RNA (siRNA) and NLRP3 inhibitor MCC950 were administered. In MIRI rats, luteolin significantly improved cardiac function and preserved endothelial barrier integrity. These effects were associated with upregulation of FOXP1 and suppression of NOD-like receptor family pyrin domain containing 3 (NLRP3) inflammasome. In OGD/R-treated endothelial cells, luteolin restored barrier function and cell viability. The protective effects of luteolin were abolished after FOXP1 silencing. Pharmacological NLRP3 inhibition (MCC950) mirrored luteolin’s protection. Our study indicates that luteolin enhances endothelial barrier function and attenuates MIRI via the FOXP1-NLRP3 pathway. The current study provides a potential drug for MIRI treatment. Full article

Cancer immunotherapy increasingly relies on nucleic acid-based vaccines, yet achieving efficient and safe delivery remains a critical limitation. Polyplexes—electrostatic complexes of cationic polymers and nucleic acids—have emerged as versatile carriers offering greater chemical tunability and multivalent targeting capacity compared to lipid nanoparticles, with lower immunogenicity than viral vectors. This review summarizes key design principles governing polyplex performance, including polymer chemistry, architecture, and assembly route—emphasizing microfluidic fabrication for improved size control and reproducibility. Mechanistically, effective systems support stepwise delivery: tumor targeting, cellular uptake, endosomal escape (via proton-sponge, membrane fusion, or photochemical disruption), and compartment-specific cargo release. We discuss therapeutic applications spanning plasmid DNA, siRNA, miRNA, mRNA, and CRISPR-based editing, highlighting preclinical data across multiple tumor types and early clinical evidence of on-target knockdown in human cancers. Particular attention is given to physiological barriers and engineering strategies—including size-switching systems, charge-reversal polymers, and tumor-penetrating peptides—that improve intratumoral distribution. However, significant challenges persist, including cationic toxicity, protein corona formation, manufacturing variability, and limited clinical responses to date. Current evidence supports polyplexes as a modular platform complementary to lipid nanoparticles in selected oncology indications, though realizing this potential requires continued optimization alongside rigorous translational development. Full article