Project description:Proteomic analysis of EVs produced by DCs differentiated in vitro from blood monocytes isolated from control, initial PD and advanced PD
Project description:<p>Blood plays a vital role in diagnostic medicine by providing essential information for assessing health and diagnosing various conditions. Routine blood tests can reveal important biomarkers that support early diagnosis and timely intervention. Recently, extracellular vesicles (EVs) have emerged as promising biomarkers due to their role in intercellular communication and involvement in disease processes such as cancer and neurodegeneration. However, isolating EVs from plasma remains challenging, as residual blood cells, especially platelets, can interfere with vesicle release. In this study, we explore the use of the PRotein Organic Solvent PRecipitation (PROSPR) method, a straightforward and effective technique previously applied to plasma and brain samples, to isolate EVs directly from whole blood. We performed detailed analyses, including ultrastructural imaging, cytometry, and multi-omics profiling, to characterize the EVs obtained. Our findings demonstrate that PROSPR can successfully capture even the smallest exosomes (30–120 nm), which are often missed in plasma-based isolation. This approach represents a significant advancement in EV research and may enhance the identification of blood-based biomarkers for a range of diseases.</p>
Project description:Venous thromboembolism (VTE) is a major contributor to morbidity and mortality in patients with advanced lung adenocarcinoma. Extracellular vesicles (EVs) are established mediators of the associated hypercoagulable state. We conducted a transcriptomic analysis of plasma EVs to identify molecular signatures specific to cancer-associated thrombosis (CAT) for refined risk stratification and therapeutic targeting. Methods: Plasma EVs from lung adenocarcinoma patients with (n=10) and without (n=11) VTE were analyzed by RNA-Sequencing. Differentially Expressed Genes (DEGs) were analyzed for pathway enrichment. Results: RNA-Seq identified 483 DEGs in VTE patients, with significant enrichment in pathways related to inflammation, neutrophil degranulation (NETs), and blood coagulation. Conclusion: Plasma EVs carry a potent, multi-cellular derived immuno-thrombotic mRNA signature that distinctly underlies CAT in lung adenocarcinoma.
Project description:Human pancreatic islets were isolated from pancreas of deceased donors by Ricordi's procedure and cultured in CMRL 1066 medium additioned with human albumin. EVs were isolated from conditioned medium derived from islet culture after isolation. Once isolated, RNA of islets and islet-derived EVs was extracted and analyzed for microRNA expression within 48 hours after isolation.
Project description:Blood plays a vital role in diagnostic medicine by providing essential information for assessing health and diagnosing various conditions. Routine blood tests can reveal important biomarkers that support early diagnosis and timely intervention. Recently, extracellular vesicles (EVs) have emerged as promising biomarkers due to their role in intercellular communication and involvement in disease processes such as cancer and neurodegeneration. However, isolating EVs from plasma remains challenging, as residual blood cells, especially platelets, can interfere with vesicle release. In this study, we explore the use of the PRotein Organic Solvent PRecipitation (PROSPR) method, a straightforward and effective technique previously applied to plasma and brain samples, to isolate EVs directly from whole blood. We performed detailed analyses, including ultrastructural imaging, cytometry, and multi-omics profiling, to characterize the EVs obtained. Our findings demonstrate that PROSPR can successfully capture even the smallest exosomes (30–120 nm), which are often missed in plasma-based isolation. This approach represents a significant advancement in EV research and may enhance the identification of blood-based biomarkers for a range of diseases.
Project description:Identification of transcriptional profile of several genes involved in diabetes in islet-derived extracellular vesicles (Evs). Recently, EVs are identified as a new mechanism in cell-to-cell communication by transfer of protein and genic information (mRNA, microRNA). Their role is under investigation in immunology, stem cell and cancer, but not in islets and diabetes. The aim of this experiment is to identify mRNA transcripts (in particular, mRNA transcripts involved in diabetes pathophysiology) present in islet Evs.
Project description:To identify specific miRNAs carried by hUCMSC-EVs, and explore their crucial roles in hUCMSC-EV-based improvement of inflammatory diseases.
Project description:As a part of study to characterize the effects of fluid flow shear stress to mouse muscle cells, small RNA sequencing was performed with muscle cell-derived extracellular vesicles (Myo-EVs).
