Project description:Myeloid-derived suppressor cells (MDSCs) are immature myeloid cells that accumulate in the tumor microenvironment of most cancer patients. There MDSCs suppress both adaptive and innate immune responses, hindering immunotherapies. Moreover, many cancers are accompanied by inflammation, a processes that further intensifies MDSC suppressive activity, causing aggressive tumor progression and metastasis. MDSCs collected from tumor-bearing mice profusely release nano-scale membrane-bound extracellular vesicles, called exosomes, which carry biologically active proteins between cells and contribute directly to the immune suppressive functions of MDSC. Many studies on other cell types have shown that exosomes may also carry microRNAs (miRNAs) and messenger RNAs (mRNAs) which can also be transferred to surrounding and distant cells. However, to the best of our knowledge, the miRNA and mRNA cargo of MDSC-derived exosomes has not yet been interrogated. This study aims to identify and quantify the cargo of MDSC and their immunosuppressive exosomes to gather knowledge that can offer insights on the mechanisms by which MDSCs contribute to immune suppression, focusing on the role of exosomes as intercellular communication mediators in the tumor microenvironment. In order to achieve our objective a well-established mouse model based on a conventional mammary carcinoma (4T1 cells) and heightened inflammation (4T1 transduced to express the cytokine interleukin-1b) was used. We provide evidence that MDSC-derived exosomes carry proteins, mRNAs and miRNAs. Relative quantitation demonstrated quantitative differences between the exosome cargo and the cargo of their parental cells, supporting the hypothesis that selective loading into the exosomes is possible. Additionally, quantitative and functional analyses of the exosome cargo generated under conventional and heightened inflammation conditions are consistent with clinical observations that inflammation is linked to cancer development.
Project description:827 human miRNA standard human panel Plasma derived extracellular vesicles (EV)/exosomes can serve as markers of cell damage/disease but can also have therapeutic utility depending on the nature of their cargo, such as miRNA. Currently there are challenges and lack of innovations regarding early diagnosis and therapeutic options within different aspects of management of trauma patients. We characterized the miRNA cargo of exosomes purified from trauma patients, and compared it to those from healthy participants. Methods Plasma was collected from a prospective, cohort study of trauma patients (n = 15; 7 males, 8 females) with hip and/or femur fractures and an injury severity score (ISS) ≥ 15; elective total hip arthroplasty (THA) patients (n = 8; 4 males, 4 females) served as operative controls. Exosomes were isolated from plasma with the Invitrogen Total Exosome Isolation Kit and RNA was isolated using a miRNeasy Mini Kit. Direct quantification of miRNA copies was performed by NanoString Technologies on a human miRNA gene panel and analyzed with nSolver Results Trauma plasma-derived exosomes had 60 miRNA that were significantly downregulated and 3 miR upregulated when compared to THA (p<0.05). Thirteen miR have a direct role in hematopoiesis regulation, including miR-223 and miR-451a. Further, male trauma plasma-derived exosomes demonstrated downregulation of 150 miR compared to male THA (p<0.05). While female trauma plasma-derived exosomes demonstrated downregulation of only four miR and upregulation of 2 miR compared to female THA (p<0.05). Conclusions Sex-specific differences in the miR from plasma-derived exosomes occurred following severe trauma. These miR changes in plasma-derived exosomes likely contribute to postinjury systemic inflammation, immune system dysregulation, and bone marrow dysfunction and may merit consideration as future potential therapeutic targets.
Project description:827 human miRNA standard human panel Plasma derived extracellular vesicles (EV)/exosomes can serve as markers of cell damage/disease but can also have therapeutic utility depending on the nature of their cargo, such as miRNA. Currently there are challenges and lack of innovations regarding early diagnosis and therapeutic options within different aspects of management of patients suffering from chronic pancreatitis (CP). Use of exosomes as biomarkers for pancreatic health, and/or or as adjuvant therapy would make a difference in management of these patients. To explore the feasibility of this approach, we characterized the miRNA cargo of exosomes purified from CP patients, and compared it to those from healthy participants. Methods: EVs were isolated from plasma of 15 CP patients and 10 healthy controls. Nanoparticle Tracking Analysis was used to determine frequency and size while NanoString technology was used to characterize the miRNA cargo. Relevant clinical parameters were correlated with these EV/exosome characteristics. Results: ~30 miRNA species were identified to have significantly (p<0.05) different expression in exosomes from individuals with CP compared to healthy individuals; ~40 miRNA were differentially expressed in exosomes from pre-diabetic versus non-diabetic CP patients. miR-579-3p, while exhibiting significantly lower (~16-fold) expression in exosomes from CP compared to healthy individuals and lower (~24-fold) in CP narcotic users compared to the less severe CP in non-users, is actually enriched (~32-fold) within exosomes in pre-diabetic CP patients compared to non-diabetic CP patients. A unique pattern was identified in female CP patients. Conclusions: These first of a kind data support the prospect of using a bioinformatics approach to assess pancreatic health, and their therapeutic potential in CP patients.
Project description:The aim of the study was to investigate whether the trefoil peptide genes, in concerted action with a miRNA regulatory network, were contributing to nutritional maintrenance. Using a Tff2 knock-out mouse model, 48 specific miRNAs were noted to be significantly deregulated when compared to the wild type strain.
Project description:The aim of the study was to investigate whether the trefoil peptide genes, in concerted action with a miRNA regulatory network, were contributing to nutritional maintrenance. Using a Tff3 knock-out mouse model, 21 specific miRNAs were noted to be significantly deregulated when compared to the wild type strain.
Project description:We investigated NDEs from depressed and psychiatrically healthy controls, before and after, eight weeks of antidepressant treatment (ADT). Neuronal-derived exosomes contained miRNA that showed enrichment for brain functions. Changes in NDE cargo, specifically miR-21-5p, miR-30d-5p, and miR-486-5p expression, were significantly associated with ADT response. Furthermore, we found these targets to be altered in brain tissue from depressed individuals. Together, our study indicates that changes in peripherally isolated NDE can act as a biomarker of ADT response specifically through size and cargo.
Project description:SILAC based protein correlation profiling using size exclusion of protein complexes derived from Mus musculus tissues (Heart, Liver, Lung, Kidney, Skeletal Muscle, Thymus)
Project description:Cancer-associated fibroblast (CAF) promote immune suppression in the tumor microenvironment by polarizing monocytes to myeloid-derived suppressor cells (MDSC) that potently suppress T-cell function. Using a patient-derived coculture system, we found that extracellular vesicles (EV) released by CAF-induced MDSC directly suppress T-cell proliferation. In this study, we profiled the protein cargo of the EVs isolated from CAF-MDSC and compared to related cell types.
Project description:SILAC based protein correlation profiling using size exclusion of protein complexes derived from seven Mus musculus tissues (Heart, Brain, Liver, Lung, Kidney, Skeletal Muscle, Thymus)