Surface plasmon resonance (SPR) biosensors are an advanced optical biosensing technology that has been widely used in molecular biology for the investigation of biomolecular interactions and in bioanalytics for the detection of biological species. This work aims to review progress in the development of SPR biosensors for medical diagnostics, focusing mainly on advances in optical platforms and assays enabling analysis of complex biological matrices. Applications of SPR biosensors for the detection of medically relevant analytes, such as nucleic acids, proteins, exosomes, viruses, bacteria, and circulating tumor cells, are also reviewed. The detection performance of current SPR biosensors is discussed, and routes for improving performance and expanding applications of SPR biosensors in medical diagnostics are outlined.
- Klíčová slova
- Biomarker, Medical diagnostics, Optical biosensor, Plasmonic affinity biosensor, Surface plasmon resonance biosensor,
- MeSH
- Bacteria izolace a purifikace MeSH
- biosenzitivní techniky * metody přístrojové vybavení MeSH
- design vybavení MeSH
- exozómy chemie MeSH
- lidé MeSH
- nukleové kyseliny analýza izolace a purifikace MeSH
- povrchová plasmonová rezonance * metody přístrojové vybavení MeSH
- proteiny analýza MeSH
- viry izolace a purifikace MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
- Názvy látek
- nukleové kyseliny MeSH
- proteiny MeSH
The critical role of the immune system in brain function and dysfunction is well recognized, yet development of immune therapies for psychiatric diseases has been slow due to concerns about iatrogenic immune deficiencies. These concerns are emphasized by the lack of objective diagnostic tools in psychiatry. A promise to resolve this conundrum lies in the exploitation of extracellular vesicles (EVs) that are physiologically produced or can be synthetized. EVs regulate recipient cell functions and offer potential for EVs-based therapies. Intranasal EVs administration enables the targeting of specific brain regions and functions, thereby facilitating the design of precise treatments for psychiatric diseases. The development of such therapies requires navigating four dynamically interacting networks: neuronal, glial, immune, and EVs. These networks are profoundly influenced by brain fluid distribution. They are crucial for homeostasis, cellular functions, and intercellular communication. Fluid abnormalities, like edema or altered cerebrospinal fluid (CSF) dynamics, disrupt these networks, thereby negatively impacting brain health. A deeper understanding of the above-mentioned four dynamically interacting networks is vital for creating diagnostic biomarker panels to identify distinct patient subsets with similar neuro-behavioral symptoms. Testing the functional pathways of these biomarkers could lead to new therapeutic tools. Regulatory approval will depend on robust preclinical data reflecting progress in these interdisciplinary areas, which could pave the way for the design of innovative and precise treatments. Highly collaborative interdisciplinary teams will be needed to achieve these ambitious goals.
- Klíčová slova
- extracellular vesicle-based therapies, extracellular vesicles, immune system, neurological and psychiatric disorders, pharmacodynamics, pharmacokinetics, regulatory agencies,
- MeSH
- biologické markery MeSH
- duševní poruchy * terapie imunologie metabolismus MeSH
- extracelulární vezikuly * imunologie metabolismus transplantace MeSH
- individualizovaná medicína * metody MeSH
- lidé MeSH
- mezibuněčná komunikace MeSH
- mozek imunologie metabolismus MeSH
- neuroglie * metabolismus imunologie MeSH
- neurony * metabolismus imunologie MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
- Názvy látek
- biologické markery MeSH
There is growing interest in the role of extracellular vesicles (EVs) in neonatal pathology. This study aimed to characterise circulating EVs following preterm birth. This single-centre prospective observational study included cord and postnatal plasma from preterm (n = 101) and full-term infants (n = 66). EVs were analysed using nanoparticle tracking analysis, flow cytometry, proteomics and procoagulant activity assay. We found changes in the concentration, size, cellular origin and proteomic content of circulating EVs in preterm infants during perinatal adaptation. To understand if these changes were related to prematurity or normal adaptation to extrauterine life, they were also investigated in term infants. There was a dramatic increase in the concentration of small and large EVs on Day 3 in the preterm group; specific subsets of platelet (CD42b+ and CD62P+), endothelial (VEGFR2) and tissue factor EVs were elevated. Differentially expressed proteins relating to haemostasis, pulmonary physiology and immunity were identified between Day 1 and 3 in preterm infants. These changes have never previously been described in a large cohort of preterm infants and differ from healthy term infants. These findings have major implications for future neonatal EV studies, particularly the timing of sample collection. Further work is required to understand the clinical implications of this unique EV profile following preterm birth.
- Klíčová slova
- haemorrhage, neonatal, umbilical cord, vascular biology,
- MeSH
- extracelulární vezikuly * metabolismus MeSH
- fyziologická adaptace * MeSH
- lidé MeSH
- novorozenec nedonošený * krev MeSH
- novorozenec MeSH
- prospektivní studie MeSH
- proteomika metody MeSH
- těhotenství MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- novorozenec MeSH
- těhotenství MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- pozorovací studie MeSH
Colorectal cancer (CRC) ranks as the second most prevalent malignancy globally, highlighting the urgent need for more effective diagnostic and therapeutic strategies, as well as a deeper understanding of its molecular basis. Extensive research has demonstrated that cells actively secrete extracellular vesicles (EVs) to mediate intercellular communication at both proximal and distal sites. In this study, we conducted a comprehensive analysis of the RNA content of small extracellular vesicles (sEVs) secreted into the culture media of five frequently utilised CRC cell lines (RKO, HCT116, HCT15, HT29, and DLD1). RNA sequencing data revealed significant insights into the RNA profiles of these sEVs, identifying nine protein-coding genes and fourteen long non-coding RNA (lncRNA) genes that consistently ranked among the top 30 most abundant across all cell lines. Notably, the genes found in sEVs were highly similar among the cell lines, indicating a conserved molecular signature. Several of these genes have been previously documented in the context of cancer biology, while others represent novel discoveries. These findings provide valuable insights into the molecular cargo of sEVs in CRC, potentially unveiling novel biomarkers and therapeutic targets.
- Klíčová slova
- CRC, Cell line, Colorectal cancer, EV, EV cargo, EV content, Exosomes, Extracellular vesicles,
- MeSH
- extracelulární vezikuly * metabolismus genetika MeSH
- HCT116 buňky MeSH
- kolorektální nádory * genetika patologie metabolismus MeSH
- lidé MeSH
- nádorové biomarkery genetika metabolismus MeSH
- nádorové buněčné linie MeSH
- regulace genové exprese u nádorů MeSH
- RNA dlouhá nekódující genetika MeSH
- sekvenční analýza RNA MeSH
- stanovení celkové genové exprese MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- nádorové biomarkery MeSH
- RNA dlouhá nekódující MeSH
Extracellular vesicles (EVs) are emerging as critical mediators of intercellular communication in the tumor microenvironment (TME), profoundly influencing cancer progression. These nano-sized vesicles, released by both tumor and stromal cells, carry a diverse cargo of proteins, nucleic acids, and lipids, reflecting the dynamic cellular landscape and mediating intricate interactions between cells. This review provides a comprehensive overview of the biogenesis, composition, and functional roles of EVs in cancer, highlighting their significance in both basic research and clinical applications. We discuss how cancer cells manipulate EV biogenesis pathways to produce vesicles enriched with pro-tumorigenic molecules, explore the specific contributions of EVs to key hallmarks of cancer, such as angiogenesis, metastasis, and immune evasion, emphasizing their role in shaping TME and driving therapeutic resistance. Concurrently, we submit recent knowledge on how the cargo of EVs can serve as a valuable source of biomarkers for minimally invasive liquid biopsies, and its therapeutic potential, particularly as targeted drug delivery vehicles and immunomodulatory agents, showcasing their promise for enhancing the efficacy and safety of cancer treatments. By deciphering the intricate messages carried by EVs, we can gain a deeper understanding of cancer biology and develop more effective strategies for early detection, targeted therapy, and immunotherapy, paving the way for a new era of personalized and precise cancer medicine with the potential to significantly improve patient outcomes.
- Klíčová slova
- Biomarkers, Cancer, Extracellular vesicles, Immune evasion, Liquid biopsy, Metastasis, Targeted therapy, Tumor microenvironment,
- MeSH
- extracelulární vezikuly * metabolismus MeSH
- lidé MeSH
- mezibuněčná komunikace * MeSH
- nádorové biomarkery metabolismus MeSH
- nádorové mikroprostředí * MeSH
- nádory * metabolismus patologie terapie MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
- Názvy látek
- nádorové biomarkery MeSH
The malaria parasite, Plasmodium falciparum, secretes extracellular vesicles (EVs) to facilitate its growth and to communicate with the external microenvironment, primarily targeting the host's immune cells. How parasitic EVs enter specific immune cell types within the highly heterogeneous pool of immune cells remains largely unknown. Using a combination of imaging flow cytometry and advanced fluorescence analysis, we demonstrated that the route of uptake of parasite-derived EVs differs markedly between host T cells and monocytes. T cells, which are components of the adaptive immune system, internalize parasite-derived EVs mainly through an interaction with the plasma membrane, whereas monocytes, which function in the innate immune system, take up these EVs via endocytosis. The membranal/endocytic balance of EV internalization is driven mostly by the amount of endocytic incorporation. Integrating atomic force microscopy with fluorescence data analysis revealed that internalization depends on the biophysical properties of the cell membrane rather than solely on molecular interactions. In support of this, altering the cholesterol content in the cell membrane tilted the balance in favor of one uptake route over another. Our results provide mechanistic insights into how P. falciparum-derived EVs enter into diverse host cells. This study highlights the sophisticated cell-communication tactics used by the malaria parasite.
- Klíčová slova
- EVs, cellular uptake, extracellular vesicles, imaging flow cytometry, malaria, membrane deformability,
- MeSH
- buněčná membrána * metabolismus MeSH
- endocytóza MeSH
- extracelulární vezikuly * metabolismus MeSH
- lidé MeSH
- monocyty metabolismus imunologie MeSH
- Plasmodium falciparum * metabolismus MeSH
- T-lymfocyty metabolismus imunologie MeSH
- tropická malárie parazitologie imunologie metabolismus MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Myocardial dysfunction, characterized by impaired cardiac muscle function, arises from diverse etiologies, including coronary artery disease, myocardial infarction, cardiomyopathies, hypertension, and valvular heart disease. Recent advancements have highlighted the roles of exosomes and non-coding RNAs in the pathophysiology of myocardial dysfunction. Exosomes are small extracellular vesicles released by cardiac and other cells that facilitate intercellular communication through their molecular cargo, including ncRNAs. ncRNAs are known to play critical roles in gene regulation through diverse mechanisms, impacting oxidative stress, fibrosis, and other factors associated with myocardial dysfunction. Dysregulation of these molecules correlates with disease progression, presenting opportunities for therapeutic interventions. This review explores the mechanistic interplay between exosomes and ncRNAs, underscoring their potential as biomarkers and therapeutic agents in myocardial dysfunction. Emerging evidence supports the use of engineered exosomes and modified ncRNAs to enhance cardiac repair by targeting signaling pathways associated with fibrosis, apoptosis, and angiogenesis. Despite promising preclinical results, delivery, stability, and immunogenicity challenges remain. Further research is needed to optimize clinical translation. Understanding these intricate mechanisms may drive the development of innovative strategies for diagnosing and treating myocardial dysfunction, ultimately improving patient outcomes.
- Klíčová slova
- Bioengineering, Cardiomyopathies, Exosomes, Myocardial dysfunction, Non-coding RNAs,
- MeSH
- biologické markery metabolismus MeSH
- exozómy * metabolismus genetika MeSH
- fibróza MeSH
- kardiomyopatie genetika patofyziologie metabolismus MeSH
- lidé MeSH
- myokard metabolismus patologie MeSH
- nekódující RNA * genetika MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
- Názvy látek
- biologické markery MeSH
- nekódující RNA * MeSH
Extracellular vesicles (EVs) from parasites have been identified as potent modulators of host-parasite interactions. However, their biogenesis and secretory activity are still poorly understood. Here we present a comprehensive examination of the secretory dynamics of two distinct EV fractions isolated from the adult tapeworm Hymenolepis diminuta. Additionally, we perform a detailed analysis of changes in proteomic content and morphology during EV secretion, utilising electron tomography to shed light on a previously described novel mechanism of EV biogenesis via bead-like protrusion. Our findings reveal a significant decrease in EV secretion between 24 and 48 h of in vitro cultivation when external host stimuli are no longer present. Finally, this study addresses, for the first known time, the potential bias in EV analysis resulting from extended in vitro cultivation of model parasites.
- Klíčová slova
- ESP, Electron microscopy, Exosomes, LC–MS/MS, Proteins, Secretion, Size exclusion chromatography,
- MeSH
- extracelulární vezikuly * metabolismus ultrastruktura MeSH
- Hymenolepis diminuta * fyziologie růst a vývoj metabolismus ultrastruktura MeSH
- interakce hostitele a parazita * MeSH
- proteomika MeSH
- tomografie elektronová MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
BACKGROUND: Despite secondary prevention with aspirin, patients with stable cardiovascular disease (CVD) remain at elevated long-term risk of major adverse cardiovascular events. The Cardiovascular Outcomes in People Using Anticoagulant Strategies (COMPASS) double-blind, randomized clinical trial demonstrated that aspirin plus low-dose rivaroxaban (COMPASS regime) significantly decreased the incidence of major adverse cardiovascular events by 24% compared with aspirin alone. However, the mechanisms underlying these potential synergistic/nonantithrombotic effects remain elusive. Extracellular vesicles (EVs) are crucial messengers regulating a myriad of biological/pathological processes and are highly implicated in CVD. OBJECTIVES: We hypothesized that circulating EV profiles reflect the cardioprotective properties of the COMPASS regime. METHODS: A cohort of stable CVD patients (N = 40) who participated in the COMPASS trial and were previously randomized to receive aspirin were prospectively recruited and assigned a revised regimen of open-label aspirin plus rivaroxaban. Blood samples were obtained at baseline (aspirin only) and 6-month follow-up. Plasma EV concentration, size, and origin were analyzed by nanoparticle tracking analysis and flow cytometry. EVs were enriched by ultracentrifugation for proteomic analysis. RESULTS: The COMPASS regime fundamentally altered small (<200 nm) and large (200-1000 nm) EV concentration and size compared with aspirin alone. Crucially, levels of platelet-derived and myeloperoxidase-positive EVs became significantly decreased at follow-up. Comparative proteomic characterization further revealed a significant decrease in highly proinflammatory protein expression at follow-up. CONCLUSION: The observed changes in EV subpopulations, together with the differential protein expression profiles, suggest amelioration of an underlying proinflammatory and prothrombotic state upon dual therapy, which may be of clinical relevance toward understanding the fundamental mechanism underlying the reported superior cardiovascular outcomes associated with this antithrombotic regimen.
- Klíčová slova
- aspirin, cardiovascular disease, extracellular vesicles, proteomics, rivaroxaban,
- MeSH
- Aspirin * aplikace a dávkování škodlivé účinky MeSH
- biologické markery krev MeSH
- časové faktory MeSH
- dvojitá slepá metoda MeSH
- extracelulární vezikuly * metabolismus účinky léků MeSH
- fibrinolytika * aplikace a dávkování škodlivé účinky MeSH
- inhibitory agregace trombocytů * aplikace a dávkování škodlivé účinky MeSH
- inhibitory faktoru Xa * aplikace a dávkování škodlivé účinky MeSH
- kardiovaskulární nemoci * krev farmakoterapie diagnóza MeSH
- kombinovaná farmakoterapie MeSH
- lidé středního věku MeSH
- lidé MeSH
- mediátory zánětu * krev MeSH
- prospektivní studie MeSH
- proteomika MeSH
- rivaroxaban * aplikace a dávkování škodlivé účinky terapeutické užití MeSH
- senioři MeSH
- trombóza * krev prevence a kontrola MeSH
- výsledek terapie MeSH
- Check Tag
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- randomizované kontrolované studie MeSH
- Názvy látek
- Aspirin * MeSH
- biologické markery MeSH
- fibrinolytika * MeSH
- inhibitory agregace trombocytů * MeSH
- inhibitory faktoru Xa * MeSH
- mediátory zánětu * MeSH
- rivaroxaban * MeSH
In recent years, there has been a growing interest in plant extracellular vesicles (pEVs) due to their immense potential for medical applications, particularly as carriers for drug delivery. To use the benefits of pEVs in the future, it is necessary to identify methods that facilitate their production in sufficient quantities while maintaining high quality. In this study, a comparative analysis of yields of tobacco pEV derived from apoplastic fluid, sterile calli, and suspension cultures, was performed to identify the most suitable plant material for vesicle isolation. Subsequent experiments focused on assessing the efficiency of small interfering RNA (siRNA) loading into callus-derived vesicles, employing various methods such as sonication, incubation, incubation supplemented with saponin, lipofection, and electroporation. Differences in loading efficiency among vesicles derived from apoplastic fluid, calli, and suspension cultures were observed. Moreover, our investigation extended to the presence of tobacco secondary metabolites, specifically anabasine and nicotine, within vesicles originating from three distinct tobacco sources. The outcomes of our study highlight variations not only in vesicle yields based on their source but also in their loadability and the presence of nicotine and anabasine. These findings contribute valuable insights into optimizing the production and application of pEVs for future medicinal purposes.
- MeSH
- buněčné kultury metody MeSH
- extracelulární vezikuly * metabolismus MeSH
- malá interferující RNA metabolismus genetika MeSH
- nikotin MeSH
- tabák * metabolismus MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- malá interferující RNA MeSH
- nikotin MeSH
