Background: Sepsis confers persistently high mortality, with risk escalating markedly upon progression to disseminated intravascular coagulation (DIC). Although circulating extracellular vesicles (EVs) have emerged as potential biomarker reservoirs and pathogenic mediators in sepsis-associated DIC (SA-DIC), the molecular mechanisms governing the transition from sepsis through sepsis-induced coagulopathy (SIC) to SA-DIC remain elusive. Methods: Plasma EVs from healthy controls (n=3) and patients with sepsis, SIC, and SA-DIC (n=6) were collected longitudinally and profiled by multi-omics analysis (miRNome, proteome, and metabolome) using RNA sequencing and LC-MS/MS. Key findings were validated by qRT-PCR and ELISA in an independent longitudinal cohort (n=30). The role of miR-34a-5p in lipopolysaccharide (LPS)induced inflammatory and procoagulant responses was investigated in human umbilical vein endothelial cells (HUVECs) using miRNA agomirs and antagomirs. In a cecal ligation and puncture (CLP)-induced SA-DIC mouse model, miR-34a-5p agomirs/ antagomirs were administered, and coagulation and inflammatory parameters, together with the histopathology of the lung, liver, and intestine, were assessed. Results: Integrated multi-omics analysis identified 112 miRNAs, 525 proteins, and 60 metabolites that were differentially expressed in plasma EVs during sepsis progression, which were enriched in blood coagulation, platelet activation, inflammatory response, energy metabolism and vitamin metabolism. We identified miR-34a-5p as a biomarker for discriminating sepsis, SIC, and SA-DIC. In vitro, miR-34a-5p downregulation attenuated inflammation, reduced thrombomodulin secretion, and increased CD138 and heparan sulfate expression. Consistently, in vivo modulation of miR-34a-5p ameliorated coagulation dysfunction and lung, liver, and intestinal histopathological injury. Conclusion: This study provides the first integrative map of plasma EVs molecular reprogramming during sepsis progression, identifying potential diagnostic biomarkers and a candidate therapeutic target.
Skin aging is an inevitable physiological process co-driven by endogenous genetic programs and exogenous environmental factors, characterized by the progressive deterioration of skin structure and function, which manifests as wrinkles, sagging, pigmentation, and impaired barrier function. Extracellular vesicles, particularly exosomes, are nanoscale membranous vesicles actively secreted by cells, functioning as pivotal mediators of intercellular communication through the transport of diverse bioactive molecules. Exosomes derived from diverse sources, including mesenchymal stem cells, somatic cells, plants, milk, and plasma, can effectively reverse skin aging through multiple mechanisms, such as alleviating oxidative damage, promoting collagen synthesis, inhibiting excessive expression of matrix metalloproteinases (MMPs), clearing senescent cells, and improving mitochondrial function. Additionally, engineering modifications and optimized delivery systems can further enhance the targeting ability and synergistic therapeutic efficacy of exosomes. However, the clinical application of exosomes remains constrained by exosome heterogeneity, the lack of standardized isolation and production methods, limited delivery efficiency, and insufficient long-term clinical evidence regarding safety and efficacy. This review systematically summarizes the fundamental research, mechanisms, and clinical evidence of exosomes in combating skin aging. We comprehensively explore the application prospects and challenges, emphasizing the significant values of different exosomes in combating skin aging.
Introduction: Lipid nanoparticles (LNPs) have enabled the clinical translation of nucleic acid therapeutics, and their composition plays a critical role in organ targeting and route-specific delivery. Intranasal (IN) administration offers direct access to the lungs, while reducing systemic exposure. However, designing LNPs capable of overcoming respiratory tract barriers remains a challenge. Methods: We used DNA barcoding (b-DNA) for high-throughput in vivo screening of an LNP library following IN administration to investigate the impact of helper lipid identity and changes in the relative helper lipid/cholesterol composition on lung delivery. Following physicochemical characterization, pooled LNPs were administered intranasally to C57BL/6 mice, and their biodistribution was quantified by next-generation sequencing. Lead formulations were further evaluated for lung transfection in vivo and pulmonary cell transfection in vitro, cell viability, concentration-response relationships, and membrane-disruptive properties were also assessed. To better understand the discrepancies between the in vivo and in vitro performance, LNP stability was investigated in simulated nasal fluid and cell culture media. Results: DOPE-based LNPs exhibited uniform diameters of approximately 90-110 nm, whereas DSPC-and DOTAP-based formulations showed greater variability in size. While DOPE-and DSPC-LNPs displayed slightly negative zeta potential values, DOTAPformulations exhibited positive values. DSPC-LNP6 has emerged as the lead formulation for lung delivery and transfection in vivo, despite exhibiting limited transfection efficiency in pulmonary cells in vitro. Notably, the impact of simulated nasal fluid on the physicochemical properties of LNPs was comparable across formulations, suggesting that intrinsic membrane properties contribute to enhanced pulmonary transfection after IN administration. Finally, DSPC-LNP6 successfully mediated TRAIL mRNA delivery and induced tumor cell death in an experimental lung metastasis model. Conclusion: Collectively, our findings provide critical insights into the development of RNA therapeutics that target the respiratory tract.
ABSTRACT Arthrogryposis multiplex congenita (AMC) is a heterogeneous group of conditions characterized by congenital contractures in at least two different body regions. Despite increasingly widespread prenatal ultrasound screening, prenatal detection rates for AMC remain disappointingly low, with fewer than half of affected fetuses diagnosed before birth. Missed or late diagnosis limits opportunities for informed counseling, optimal delivery planning, and early postnatal intervention. In this overview, we summarize current knowledge on prenatal detection of AMC, drawing on international cohort studies, registry data, literature on fetal motor assessment, and patient‐reported experiences. We highlight the added value of systematic evaluation of fetal movements in addition to repeated structural anomaly scans, the role of multidisciplinary care pathways, and the importance of phenotypic classification (AMC Groups 1–3) for prognosis and counseling. We also review current and emerging genetic testing strategies, including exome‐ and genome‐based approaches, and their diagnostic yield in fetuses with contractures. Finally, we outline future perspectives for international collaboration, standardized data collection from fetus to adulthood, and research priorities with patient support organizations. Together, these efforts aim to improve awareness, earlier recognition, and more tailored counseling for parents confronted with contractures and the possibility of diagnosing AMC during their pregnancies.
Purpose: Although inhibition of the JAK-STAT signaling pathway has shown promise in cancer therapy, the therapeutic potential of PROTAC-mediated JAK1/JAK2 degradation in bladder cancer remains to be fully explored. To improve the delivery and therapeutic efficacy of PROTAC-mediated JAK1/JAK2 degradation in bladder cancer, this study developed PPcJ, a cRGD-functionalized PLGA- PEG nanoparticle encapsulating the PROTAC-based JAK1/JAK2 degrader JAPT8. Methods: PPcJ nanoparticles were characterized using transmission electron microscopy, dynamic light scattering, and highperformance liquid chromatography. Flow cytometry and confocal microscopy were used to evaluate cytotoxicity, anti-tumor effect, and intracellular localization, while Western blotting assessed the expression of JAK/STAT pathway and apoptosis-related proteins. In vivo tumor-associated fluorescence, antitumor efficacy, survival, histopathology, and serum biochemical parameters were assessed in tumor-bearing mice. Results: PPcJ exhibited a spherical nanoscale morphology. PPcJ achieved near-plateau cellular uptake at low concentrations and partially evaded lysosomal degradation, which significantly induced apoptosis in bladder cancer cells and suppressed key components of the JAK/STAT signaling pathway. In vivo, cRGD modification enabled higher tumor-specific accumulation and prolonged retention. PPcJ inhibited tumor growth by 86.2% and prolonged mean survival from 22.4 +/- 4.2 to 41.0 +/- 2.2 days, without significant changes in body weight, major-organ histology, or serum AST, ALT, urea, and creatinine levels under the tested conditions. Conclusion: These findings provide a preclinical proof-of-concept that the PPcJ nano-delivery system improves the delivery and antitumor performance of JAPT8. This strategy warrants further evaluation in systemic therapeutic approaches for bladder cancer.
