Nanostring analysis of exosomal miRNA from plasma of head and neck cancer (HNSCC) patients
Ontology highlight
ABSTRACT: Exosomes were isolated from plasma of n = 12 healthy donors (HD) and n = 16 head and neck cancer (HNSCC) patients. miRNA profiling of exosomes was performed using nCounter SPRINT system. miRNAs being predicted to target EMT-related genes (CDH1, VIM, TWIST1 and SNAI1/2) were selected and compared between HD and HNSCC patients.
Project description:Exosomes, extracellular vesicles of 30-150 nm, are released by normal and tumor cells. These nanovesicles play a major role in cell communication and can be found in biological fluids as carriers of biomarkers. Tumor exosomes can inhibit immune responses, mediate drug resistance and transform mesenchymal stem cells. In contrast to healthy donors, cancer patients’ plasma contain higher levels of exosomes. Cutaneous melanoma is a very aggressive cancer whose incidence has rapidly increased worldwide and the prognosis is generally poor, given the propensity of melanoma cells to spread to distant sites while evading immune system control.We investigated potential differences between plasma exosomes derived from healthy donors (HD) and melanoma patients at 0-I, II, and III-IV stages of disease.
Project description:Despite a significant progress in the treatment of Acute Respiratory Distress Syndrome (ARDS), our ability to identify early patients and predict outcome remains limited. In this study, we aimed to characterize small RNA content of plasma exosomes from ARDS patients in order to identify potential diagnostic biomarkers of the disease. For the first time, we profiled miRNA expression levels in plasma-derived exosomes from ARDS patients (n=8) compared to healthy subjects (n=10) by small RNA-seq. It allowed us to identify 12 exosomal miRNAs differentially expressed in ARDS context (padj<0.05).
Project description:The experiment is for demonstrating the miRNA profiles in plasma exosomes derived from mild cognitive impairment and Alzheimer's disease patients and healthy donors.
Project description:The goal of this study is to report that breast cancer associated exosomes contain microRNAs (miRNAs) associated with the RISC Loading Complex (RLC) and display cell-independent capacity to process precursor microRNAs (pre-miRNAs) into mature miRNAs. Pre-miRNAs, along with Dicer, AGO2, and TRBP, are present in exosomes of cancer cells. CD43 mediates the accumulation of Dicer specifically in cancer exosomes. Cancer exosomes mediate an efficient and rapid silencing of mRNAs to reprogram the target cell transcriptome. Exosomes derived from cells and sera of patients with breast cancer instigate non-tumorigenic epithelial cells to form tumors in a Dicer-dependent manner. These findings offer opportunities for the development of exosomes based biomarkers and therapies. Exosomes from cancer cells and non-tumorigenic cells were isolated using established ultracentrifugation methods. The global miRNA content of cancer exosomes and normosomes was investigated. Profiling of cells themselves was also used as a control. Exosomes with Dicer down regulation (MCF10AshDicer and MDA-MB-231shDicer exosomes), as well as MDA-MB-231 exosomes that contain a Dicer antibody inside were used to study the function Dicer protein in the microRNA biogenesis in exosomes.
Project description:The goal of this study is to report that breast cancer associated exosomes contain microRNAs (miRNAs) associated with the RISC Loading Complex (RLC) and display cell-independent capacity to process precursor microRNAs (pre-miRNAs) into mature miRNAs. Pre-miRNAs, along with Dicer, AGO2, and TRBP, are present in exosomes of cancer cells. CD43 mediates the accumulation of Dicer specifically in cancer exosomes. Cancer exosomes mediate an efficient and rapid silencing of mRNAs to reprogram the target cell transcriptome. Exosomes derived from cells and sera of patients with breast cancer instigate non-tumorigenic epithelial cells to form tumors in a Dicer-dependent manner. These findings offer opportunities for the development of exosomes based biomarkers and therapies. Exosomes from cancer cells were isolated using established ultracentrifugation methods. The global miRNA content of non-tumorigenic cells was investigated before and after exosomes treatment to study the role of microRNA biogenesis in exosomes for cancer progression and the transformation process of normal cells.
Project description:Signalling between endothelial cells, endothelial progenitor cells and stromal cells is crucial for the establishment and maintenance of vascular integrity and involves exosomes, among other signalling pathways. Exosomes are important mediators of intercellular communication in immune signalling, tumour survival, stress responses and angiogenesis. The ability of exosomes to incorporate and transfer mRNAs encoding for ‘acquired’ proteins or miRNAs repressing ‘resident’ mRNA translation suggests that they can influence the physiological behaviour of recipient cells. We here demonstrate that miR-214, a miRNA that controls endothelial cell function and angiogenesis, plays a dominant role in exosome-mediated signalling between endothelial cells. Endothelial cell-derived exosomes stimulated migration and angiogenesis in recipient cells, whereas exosomes from miR-214 depleted endothelial cells failed to stimulate these processes. Exosomes containing miR-214 repressed the expression of Ataxia Telangiectasia Mutated in recipient cells, thereby preventing senescence and allowing blood vessel formation. Concordantly, specific reduction of miR-214 content in exosome-producing endothelial cells abolishes the angiogenesis the angiogenesis stimulatory function of the resulting exosomes. Collectively our data indicate that endothelial cells release miR-214 containing exosomes to stimulate angiogenesis through silencing of Ataxia Telangiectasia Mutated in neighbouring target cells. Gene expression analysis of HMEC endothelial cells exposed to supernatant containing either HMEC derived exosomes (miR-214 high), HMEC derived exosomes depleted of miR-214 (miR-214 low) or containing no exosomes (no exosomes). Each sample was analysed in duplo.
Project description:Comparing the miRNA profile of exosomes from ASML vs ASML CD44v kd pancreatic adenocarcinoma line revealed that the expression level of 33 from the 40 most abundant miRNA differed significantly between ASMLwt and ASML-CD44vkd exosomes In this study tumor cell exosomes, deemed important for intercellular communication were analysed for their miRNA content. Exosomes from a highly metastatic pancreatic adenocarcinoma cell line, ASML and from a CD44v kd of ASML were used for RNA preparation using Trizol reagent, and the total RNA was sent to Genomics core facility,EMBL, Heidelberg for miRCURY LNA microRNA Array, v.11.0 - hsa, mmu & rno
Project description:We found that cardiac fibroblasts produce and secrete exosomes. miRNA profiling and TaqMan qRT-PCR experiments identified miR-21 expression to be higher in cardiac fibroblasts compared to those of miR-21*, whereas in exosomes miR-21* expression was higher compared to miR-21. The purpose of the study was to validate these findings by miRNA sequencing in cardiac fibroblasts and fibroblasts-derived exosomes. Neonatal rat cardiac fibroblasts were cultured in DMEM + 1% exosome-depleted FBS for 48h. Conditioned medium was collected and exosomes were purified by several centrifugation and filtration steps, following ultracentrifugation. Afterwards total RNA from cardiac fibroblasts and exosomes was isolated for miRNA sequencing.
Project description:Daily transcriptomic profiling was conducted on whole blood collected from COVID-19 cases. Whole blood was collected in Tempus Blood RNA tubes, and RNA was extracted from whole blood using the Tempus Spin RNA Isolation Kit. Healthy controls consisted of participants of a measles, mumps and rubella re-vaccination study. Pre-vaccination whole blood was collected and processed and analyzed as above.
Project description:Daily transcriptomic profiling was conducted on whole blood collected from COVID-19 cases. Whole blood was collected in Tempus Blood RNA tubes, and RNA was extracted from whole blood using the Tempus Spin RNA Isolation Kit. Healthy controls consisted of participants of a measles, mumps and rubella re-vaccination study. Pre-vaccination whole blood was collected and processed and analyzed as above.