cGAS-STING signaling pathway in urogenital oncology: Regulation, resistance, and routes to response.

Exosomes, nanoscale extracellular vesicles (EVs), are secreted by diverse cell types including mesenchymal stem cells, T lymphocytes, neurons, and mast cells.Saliva is among the body fluids that contain exosomes. Saliva is gaining prominence as a diagnostic biofluid due to its non-invasive and cost-effective collection. Salivary exosomes contribute significantly to maintaining oral homeostasis, modulating immune responses, and regulating the oral microbiome. Composed of proteins, lipids, RNA, and DNA, exosomes mediate intercellular communication and transport molecular cargo between cells. Due to their biological content and specific surface markers, salivary exosomes show strong potential in diagnostics, targeted drug delivery, immunomodulation, and tissue regeneration.This manuscript explores the structural components, biological functions, isolation techniques, and analytical methods associated with salivary exosomes, highlighting their role in oral and systemic health and their translational potential in clinical applications.

Following spinal cord injury (SCI), neuroinflammation driven by lipid-laden macrophage foam cells is a key pathology, yet how these cells manage their lipid homeostasis is unclear. We delineate a neuroprotective axis in which macrophages deploy apolipoprotein E (APOE) to transfer intracellular lipids to neighboring cells, especially fibroblasts. Genetic ablation of Apoe disrupts this intercellular lipid transport, culminating in pathological lipid retention that activates the Hippo signalling cascade and transcriptionally induces complement component C1q. This excess C1q aberrantly tags intact synapses for excessive microglial pruning, leading to significant synaptic loss and impaired locomotor function recovery. Direct blockade of C1q using neutralizing antibodies recapitulated these neuroprotective effects, confirming C1q as the critical mediator. Crucially, macrophage-specific APOE re-expression reverses this entire cascade, preserving synapses and restoring locomotor function (BMS score: 4.81 ± 0.21 (Apoe) vs. 1.75 ± 1.28 (NC); Incline plane: 69.24° ± 2.33° (Apoe) vs. 51.66° ± 5.14° (NC) in Apoe−/− mice). These findings identify the APOE-Hippo-C1q pathway in macrophages as a novel therapeutic target for SCI.

The intercellular transportation of molecules is crucial for regulating cell communication and function. However, the existing techniques for molecule transfer across cell barriers often cause cellular damage or have low transfer efficiencies. To address these limitations, this study proposes an innovative nanotube membrane-based injector (nanoinjector) system capable of extracting diverse cytoplasmic molecules from source cells and transferring them to target cells. The developed system demonstrates high efficiency, with over 95% viability and 90% transfer efficiency. Additionally, it enables mitochondrial transfer, which enhances cellular adenosine triphosphate (ATP) production by up to 25% within 24 h. This study explores the impact of intracellular content transport, enabled by this new tool, on cellular activities, with promising implications for cell surgery and therapy.

Sugars are the structural building blocks of carbohydrates, which are transported through a series of transporters in plant. To explore the molecular mechanisms of how transporters, play roles in uptake, transport and mobilization of sugars in maize, a series of bioinformatics analyses were done to identify and characterize the transporters. Following the analyses, 60 sugar transporters were identified in maize, which shared eight (STP, PLT, ERD6, INT, TMT, pGlcT, SUC, and VGT) clades during phylogenetic analysis. Due to having significant differences in molecular weight, multiple beta-strands, transmembrane helices, and 11-12 transmembrane domains, the transports might play significant variations in functional properties. Since most transporters are plasma membrane bound, and have the highest homolog pairs (39) with S. bicolor during synteny analysis, the transporters might be involved in intercellular sugar transport that are conserved and significantly duplicated during the process of evolution. The lowest binding affinity (ΔG: - 7.1 kcal/mol) in ZmVGT1-Suc docked complex, and most commonly found hydrogen bond mediated attachment of valine residue ligand might represent the complex stability and functional integrity of the complex. Indeed, the RMSD deviation of 1 (for ZmST10-Gal) to 3 Å (for ZmST10-Glu) among the docked complexes might guide the subtle conformational differences that could impact the functional roles of the complexes. Next, during co-expression analysis, clustering of 491 genes with 43 maize sugar transporter into four co-expression clusters and five different metabolic pathways might guide their inter regulatory roles in interacting different metabolic pathways. More specifically, co-expression of ZmSTP9 and ZmPLT10 with the MYB8 and A6b stress-responsive transcription factors might guide their stress regulatory mechanisms. The RNA-Seq based observation of differential tissue specific expression and expression under salinity, drought, nitrogen deficiency, and heat stress and qRT-PCR mediated validation of differential tissue specific expression and upregulation of ZmPLT1, ZmPLT8, ZmSTP1, ZmTMT1, and ZmSUC3 under salinity stress might guide their potential roles in abiotic stress tolerance. The plasma membrane localized validation of subcellular localization of ZmPLT1 and ZmPLT8 proteins might guide the consistent results between dry and wet lab experiments. Therefore, the identified and characterized maize sugar transporters through integrated dry and wet lab experiments might guide the future research in developing abiotic stress tolerant maize and exploring the molecular mechanism of stress tolerance trough transporter guided regulation of maize abiotic stress signaling pathways following circuit enabled synthetic biology approaches.

Sepsis is a complex condition marked by significant dysregulation of immune and metabolic processes, leading to multi-organ failure. Macrophages, key mediators of immune activity, demonstrate functional flexibility by switching between pro- and anti-inflammatory phenotypes in response to inflammatory and metabolic signals in their local environment. During sepsis, pathogen-derived signals activate host defense responses that impair intercellular oxygen transport, increase oxygen consumption by immune cells within inflamed tissues, and promote a metabolic transition toward aerobic glycolysis. This metabolic transition supports immune defense mechanisms, and the metabolic by-products further regulate immune activation through feedback in key signaling cascades, promoting a transition toward tolerance during the resolution phase. Since mitochondria are central hubs for cellular energy homeostasis, they play a crucial role in this process. Mitochondrial dysfunction and metabolic changes are now recognized as major contributors to the progression of sepsis. The accumulation of mitochondria-derived metabolites can further modulate immune signaling pathways, actively influencing macrophage function. Therefore, this review emphasizes the crosstalk between macrophage polarization and mitochondrial changes, with a focus on new molecular insights and the potential of mitochondrial pathways as biomarkers or therapeutic targets. These concepts provide a foundation for advancing both experimental research and clinical applications, potentially guiding future interventions to better manage sepsis and its associated mortalities.

Inspired by the non-transmembrane transfer of mitochondria in cell-to-cell communications, herein, we report an original exploration to accelerate mitochondrial intercellular transport, and its application to exogenous cargo delivery. We discover that deliberate PINK1-targeted mitophagy downregulation elevates mitochondrial transit capacity via multifaceted drivers-morphological adaptation, metabolic reprogramming, and respiratory enhancement. Capitalizing on this, we engineer high-speed mitochondrial vehicles for photosensitizer hitchhiking, with spatiotemporal tracking elucidating its dynamic intercellular transit and physiological impacts. Through mitochondria's communication network-tunneling nanotubes (TNTs), the mitochondria-photosensitizer cotransporter achieves reinforced intercellular delivery, thereby inducing deep tumor penetration and enhanced photodynamic killing. Our work establishes a transformative mitochondria-hitchhiking platform for overcoming biological barriers in drug delivery and provides mechanistic insights into manipulating intercellular organelle transport for therapeutic applications.

Cell membrane-derived vesicles play essential roles in intercellular communication, material transport, and waste disposal. Despite their biomedical and industrial potential, isolating extracellular vesicles from natural sources remains technically challenging, limiting purification efficiency and scalability. This study introduces cell membrane extrusion as an alternative approach to optimize the production of cell membrane-derived vesicles (CSMs), from eukaryotic and prokaryotic cells. CSMs, generated from HeLa and SH-SY5Y cells exhibited a distinctive cup-shaped morphology and sizes of 151.36 ± 72.36 nm, and 416.86 ± 108.49 nm at 20 °C by DLS respectively, showing remarkable thermal stability at 4-70 °C range. Furthermore, loaded vesicles interacted with mammalian cells and achieved successful cargo internalization. CSMs were also produced from E. coli membranes, forming unilamellar vesicles of approximately 100 nm, as observed by Cryo-TEM. These vesicles displayed an inverse correlation between vesicle size and thermal stability and efficient cargo incorporation detected in 85% ± 3% of CSMs. However, under tested conditions, no interaction with prokaryotic cells occurred, and consequently, no delivery of the loaded molecule was observed. Overall, thesefindings highlight the potential of generating cell membrane-derived nanovesicles through extrusion, offering a promising strategy to mimic extracellular vesicles for innovative biomedical and industrial applications, including targeted drug delivery system.

Mitochondrial transfer through tunneling nanotubes inspires an innovative strategy for intercellular drug delivery.

The tomato zonate spot virus (TZSV) poses a significant threat to agriculture. Therefore, the elucidation of the functional roles and interactions of its encoded proteins is crucial for the development of effective control strategies. The aim of this study was to investigate the interaction network between the TZSV nucleocapsid (N), the non-structural M-segment (NSm) and the non-structural S-segment (NSs) proteins, with a focus on the functional characterization of the NSm protein. Yeast two-hybrid (Y2H) analysis indicated that both the N protein (N-N) and the NSm protein (NSm-NSm) exhibit self-interaction in vitro, with successful expression of all fusion proteins confirmed by Western blotting. Subsequently, we used bimolecular fluorescence complementation (BiFC) and luciferase complementation imaging (LCI) assays in epidermal cells of Nicotiana benthamiana to confirm that N and NSm proteins self-interact. In addition, heterologous interactions between NSs-N, N-NSm and NSs-NSm were also detected. BiFC and co-localization experiments with fusion proteins elucidated the interaction place of the cell: N-N and NSm-N interactions occurred in both the cytoplasm and nucleus, with NSm-NSm interaction occurring in the nucleus, whereas NSs-N and NSs-NSm interactions only occurred in the cytoplasm. Subcellular localization studies showed that the N protein is distributed in both the cytoplasm and the nucleus, whereas the NSm and NSs proteins are predominantly localized in the cytoplasm. In particular, NSm was found to specifically target plasmodesmata (PD) and co-localize with the known PD marker protein PDLP8. Interestingly, TZSV NSm was demonstrated to mediate the cell-to-cell movement of a cucumber mosaic virus mutant (ΔCMV-GFP) lacking its native movement protein (3a). This was evidenced by the spread of approximately 50 fluorescent foci to neighboring cells observed at 6 dpi. This study comprehensively describes the intricate interaction network between the N, NSm and NSs proteins of TZSV and clarifies their subcellular localizations within plant cells. Crucially, we provide conclusive evidence that the NSm protein of TZSV is a functional movement protein essential for facilitating viral intercellular transport which promotes viral spread within the host during systemic infection. These findings offer important insights into the infection mechanism of TZSV and provide potential targets for the control of TZSV.

The biological functions of the multiple members of the xyloglucan endotransglycosylase/hydrolase (XTH) protein family are rather diverse: XTHs are cell wall remodeling enzymes that participate in plant growth and development, are involved in responses to various environmental stresses and interactions with pathogenic and symbiotic microorganisms. However, XTHs’ role upon viral infection remains poorly understood. Here we identified and characterized Nicotiana benthamiana XTH (NbXTH) which is involved in responses to viral infection. We demonstrated that NbXTH is a positive regulator of intercellular transport. NbXTH suppression leads to the inhibition of tobacco mosaic virus (TMV) local spread, resulting in the increased tolerance of N. benthamiana plants to TMV. Therefore, NbXTH could be regarded as a susceptibility factor.

Glycerophospholipids (GPs) are integral constituents of cellular membranes, and play a crucial role in the regulation of lipid metabolism homeostasis and physiological conditions. However, pathological alterations associated with aging, such as variations in plasma GP concentrations, disruptions in intercellular GP transport, and local accumulation of excessive GPs, have been observed. These changes induce irreversible cellular degeneration, ultimately leading to tissue damage in organs such as the brain and retina. A growing body of evidences has demonstrated that GPs play significant roles in neurodegenerative diseases, including Alzheimer's disease (AD) and Parkinson's disease (PD). Similarly, GPs have been implicated in the pathogenesis and progression of age-related macular degeneration (AMD), a degenerative condition affecting the choroid and retinal layers of the eye. Understanding the homeostasis of GP metabolism in the aging retina and in AMD is essential for elucidating the pathogenic processes involved in AMD. In this review, we present a comprehensive overview of the mechanisms of GPs in the aging retina and their correlation with degenerative processes associated with AMD. KEY MESSAGES: What is known Metabolic dysregulation of glycerophospholipids (GPs) plays vital roles in age-related macular degeneration (AMD) and neurodegenerative diseases, such as Alzheimer's disease (AD) and Parkinson's disease (PD). Patients with age-related neurological disorders exhibit a significantly higher risk of developing AMD compared to healthy individuals, potentially due to shared pathological mechanisms, including mitochondrial metabolic disturbance, chronic inflammation and autophagy dysfunction. What is new The interconnection between multiple GP species and their metabolites has been established to delineate complex pathogenic mechanisms underlying the aging retina and AMD, including cell senescence, autophagy and apoptosis, oxidative stress, inflammation, vascular abnormalities. GPs may serve as potential therapeutic targets to prevent or delay the progression of AMD.

Extracellular vesicles are membrane-enclosed structures released by cells into the extracellular space, containing various biomolecules such as proteins, nucleic acids, and lipids. Extracellular vesicles exhibit broad cellular origins, diverse types, and high heterogeneity. They are involved in intercellular material transport, mediate intercellular communication, and play important roles in various cellular biological processes, including cell proliferation, apoptosis, and migration. This review summarizes recent advances in research on the isolation and identification, biogenesis mechanisms, and fate of extracellular vesicles, aiming to provide a reference for advancing research in this field.

Small secreted extracellular vesicles (EVs) mediate intercellular transport of bioactive macromolecules. How the membrane lipid phosphatidylinositol 4,5-bisphosphate [PI(4,5)P2], which plays a crucial role in many cellular processes, impacts EV biogenesis is unclear. The primary cilium, a sensory organelle protruding from most non-dividing cells, transmits signals by shedding EVs called ectosomes. Here, we altered ciliary PI(4,5)P2 in C. elegans by manipulating the expression of the type I phosphatidylinositol 4-phosphate 5-kinase (PIP5K1) PPK-1 and deletion of the phosphoinositide 5-phosphatase (INPP5E) inpp-1, then determined the impact on release of EVs that carried cargoes tagged with fluorescent proteins. We discovered that increasing PI(4,5)P2 differentially affected ectosome shedding from distinct compartments, decreasing biogenesis of an EV subpopulation from the ciliary base, but enhancing budding from the cilium distal tip. Altering PI(4,5)P2 levels also impacted the abundance and distribution of EV cargoes in the cilium, but not the sorting of the protein cargoes into distinct subsets of ectosomes. Finally, manipulating PI(4,5)P2 did not affect cilium length, suggesting that changing PI(4,5)P2 levels can serve as a mechanism to regulate ectosome biogenesis in response to physiological stimuli without impacting cilium morphology.

A coordinated and generalized plant response to adverse environmental factors largely depends on the proper and finely-tuned regulation of intercellular transport via plasmodesmata (PD). However, the knowledge of the whole network of PD-controlling mechanisms is far from complete. Earlier, a cellular factor, Kunitz peptidase inhibitor-like protein (KPILP), that affects PD gating and plays a proviral role, was identified in Nicotiana benthamiana plants. Here we characterized its homolog from N. tabacum, NtKPILP, which is hardly detectable in leaves of intact plants, in contrast to roots, flowers and seeds where NtKPILP is highly expressed. However, its mRNA accumulation in leaves increases in response to various stresses, including viral infection. NtKPILP was demonstrated to affect chloroplast functioning. Using the virus-induced gene silencing approach, we have shown that NtKPILP downregulation negatively affects intercellular transport of macromolecules, inducing callose deposition at PD and suppressing beta-1,3-glucanase mRNA accumulation. Together, the obtained results indicate that NtKPILP is a viral infection-responsive cellular factor that is involved in PD permeability regulation, sharing thus the features of KPILPs from other Nicotiana species.

Small secreted extracellular vesicles (EVs) mediate intercellular transport of bioactive macromolecules. How the membrane lipid phosphatidylinositol 4,5-bisphosphate (PI(4,5)P2), which plays a critical role in many cellular processes, impacts EV biogenesis is unclear. The primary cilium, a sensory organelle protruding from most non-dividing cells, transmits signals by shedding EVs called ectosomes. Here, we altered ciliary PI(4,5)P2 by manipulating expression of the type I phosphatidylinositol 4-phosphate 5-kinase (PIP5K1) PPK-1 and deletion of the phosphoinositide 5-phosphatase (INPP5E) inpp-1, then determined the impact on release of EVs that carried cargos tagged with fluorescent proteins. We discovered that increasing PI(4,5)P2 differentially affected ectosome shedding from distinct compartments, decreasing biogenesis of an EV subpopulation from the ciliary base, but enhancing budding from the cilium distal tip. Altering PI(4,5)P2 levels also impacted the abundance and distribution of EV cargos in the cilium, but not the sorting of the protein cargos into distinct subsets of ectosomes. Finally, manipulating PI(4,5)P2 did not affect cilium length, suggesting that changing PI(4,5)P2 levels can serve as a mechanism to regulate ectosome biogenesis in response to physiological stimuli without impacting cilium morphology.

Closed capsules, such as lipid vesicles, soap bubbles, and emulsion droplets, are ubiquitous throughout biology, engineered matter, and everyday life. Their creation and disintegration are defined by a singularity that separates a topologically distinct extended liquid film from a boundary-free closed shell. Such topology-changing processes are of fundamental interest. They are also essential for intercellular transport, transcellular communication, and drug delivery. However, studies of vesicle formation are challenging because of the rapid dynamics and small length scale involved. We develop fluid colloidosomes, micrometer-sized analogues of lipid vesicles. The mechanics of colloidosomes and lipid vesicles are described by the same theoretical model. We study colloidosomes close to their disk-to-sphere topological transition. Intrinsic colloidal length and time scales slow down the dynamics to reveal colloidosome conformations in real time during their assembly and disassembly. Remarkably, the lowest-energy pathway by which a closed vesicle transforms into a flat disk involves a topologically distinct cylinder-like intermediate. These results reveal aspects of topological changes that are relevant to all liquid capsules. They also provide a robust platform for the encapsulation, transport, and delivery of nanosized cargoes.

Mechano-activated connexin hemichannels mediate intercellular glutathione transport and support lens redox homeostasis.

Extracellular vesicles (EVs) are membrane-bound vesicles secreted by cells, including exosomes, microvesicles, and apoptotic bodies, which play critical roles in intercellular communication, material transport, and signal transduction. In recent years, increasing evidence has highlighted the essential function of EVs in early embryo development. By carrying bioactive molecules such as proteins, nucleic acids (e.g., mRNA and miRNA), and lipids, EVs regulate embryonic gene expression, cell proliferation, differ-entiation, and the microenvironment. Studies have shown that EVs derived from various segments of the female reproductive tract can enhance embryonic developmental potential, improve embryo quality, and facilitate implantation. Additionally, EVs secreted by embryos themselves participate in intercellular communication and play pivotal roles during embryogenesis. This review summarizes recent advances in understanding the functions of EVs in early embryo development, discusses their roles in mediating cell-to-cell com-munication and regulating gene expression, and explores the potential applications in reproductive medicine and clinical practice, offering new perspectives for optimizing assisted reproductive technologies.

Intercellular pathways of viral infection in host cells offer advantages, such as efficiency of viral spread and immune surveillance evasion, compared to cell-free viral infection. Therefore, some enveloped viruses present both cell-to-cell and cell-free forms of infection in the host organisms. In this study, we investigated the occurrence of Ebola virus (EBOV) and Marburg virus (MARV) nucleocapsid exchange in vitro between interconnected Huh7 cells using live-cell imaging methods. Moreover, through plasmid transfection methods, we demonstrated that nucleocapsid-like structures (NCLSs) formed with EBOV NP, VP35, VP24, and VP30 proteins can also be transported intercellularly to non-transfected cells through cell-to-cell contact regions in a process involving interaction with the host cell actin cytoskeleton. Our results provide further evidence of cell-to-cell transport as a mechanism of filovirus spread and support the need for further research in this field to develop new intervention methods targeting this transmission pathway.