Project description:We performed a comprehensive miRNA profiling analysis of exosomes by Treponema pallidum-stimulated microarrays. A total of 2×106 macrophages were obtained by THP-1 differentiation and grown in RPMI-1640 containing 10% exosome-free FBS. Exosomes were acquired from macrophage culture supernatants with (n = 7) or without (n = 3) T. pallidum. Briefly, macrophages were washed in PBS twice and further grown in fresh medium for 12 h (n = 2), 24 h (n = 2) and 48 h (n = 3) to collect exosomes. Exosomal miRNA microarray assays were carried out with Agilent Human miRNA (8*60K) array.
Project description:Mycobacterial transcripts were identified in exosomes released from M.tb infected RAW264.7 macrophages that were not present in uninfected exosomes suggesting export of mycobacterial RNA via exosomes Mycobacterial RNA was used as positive control and RNA from exosomes released from uninfected macrophages was used as negative control
Project description:Investigation to study mRNA transcripts present in exosomes from M.tb infected cells and how they compare to those derived from uninfected cells. Transcripts were also studied in donor macrophages as controls The gene expression study identified unique transcripts as well as differentially expressed transcripts present in exosomes released from infected macrophages
Project description:Macrophages are recognized as the vital players in renal fibrosis, with a high degree of heterogeneity and plasticity, and triggering receptor expressed on myeloid cell-2(TREM-2) was highly expressed on macrophages and participated in the progression of tissue fibrosis. However, the mechanism by which TREM-2 mediate the progress of renal fibrosis is still unclear. Our study have found the exosomes derived from TREM-2 deficient (TREM-2-/-) macrophages could suppress the progression of fibrosis, showing a higher matrix metalloproteinase-9 (MMP-9) / tissue inhibitor of matrix metalloproteinase-1 (TIMP-1) ration at protein level in secreted exosomes than exosomes from WT(wild type)macrophages in the fibrotic microenvironment. Besides, renal tubular epithelial cells (TECs) engulfed theses nanoscale vesicles, the expression of collagen I and α-smooth muscle actin (α-SMA) (fibrosis related marker) was obviously decreased. Through the RNA-seq, we found that TREM-2-/- macrophages upregulate the MMP-9/TIMP-1 ratio in its exosomes via HSPa1b/AKT pathway. Furthermore, it’s noteworthy that the renal fibrosis was effectively alleviated in the obstructed kidney from mice received a renal pelvis injection of adeno associated virus (AAV-shTREM-2) containing the sequence of silencing TREM-2. However, VER-155008 (inhibitor of HSPa1b) and Ly294002 (inhibitor of AKT) reversed this effect. Moreover, polyclonal antibodies against TREM-2 also effectively relieved the UUO-induced renal fibrosis. Above all, we have validated that the knocking down TREM-2 expression can inhibit the progression of renal fibrosis through a macrophage exosome-dependent pathway in vitro and vivo. Hence, our finding suggest TREM-2 is potential therapeutic target for CKD.
Project description:Small RNA deep sequencing analysis on the microRNA components within exosomes secreted from adipose tissue macrophages of lean and obese mice
Project description:Unprogrammed macrophage polarization, is associated with diabetic wound ulcers. Nevertheless, development of corresponding drugs is still a challenge. Here, exosomes are isolated from naive bone marrow-derived macrophages (BMDMs) (M0-Exos), inflammatory BMDMs (M1-Exos), and anti-inflammatory BMDMs (M2-Exos), with the aim of pinpointing the exosomes functionality and identify global miRNAs expression profiles.
Project description:Nasopharyngeal carcinoma is characterized by Epstein-Barr virus (EBV) infection and severe immune cell infiltration. Here, we explored the effect of tumor-derived exosomes on NPC microenvironment using single-cell RNA sequencing (scRNA-seq). We investigated the phenotypic and functional alterations in macrophages, as well as exosome-mediated cellular interactions. Human NPC biopsy samples were dissociated into single-cell suspension and stimuated with exosomes derived from an EBV positive cell line (HK1EBV) for 2 hours. A total of 14,446 qualified cells were subjected to analysis. Collectively, our findings revealed that NPC-derived exosomes facilitate immunosuppressive microenvironment and contribute to NPC progression.
Project description:To determine the different gene signatures between primary tumor and tumor-derived exosomes, we have employed RNA-sequencing as a discovery platform to identify gene signatures of tumor-derived exosomes, taking the original tumors as a control. We subcutaneously inoculated C57BL/6 mice with Lewis lung carcinoma (LLC). Three weeks later, tumor tissues were cut and tumor-derived exosomes were isolated as described in the "treatment protocol". Then, both exosomal RNA and tumor RNA were extracted and sequenced. From sequencing, we found that exosomal RNAs showed quite different transcript profiles from tumor RNAs. Examination of different gene signatures between primary tumor and tumor-derived exosomes. 2 replicates each.
Project description:Macrophages are abundant in uterine mucosa during the peri-implantation phase and early pregnancy. Decidual macrophages display dynamic changes alone with pregnancy progress: During the peri-implantation phase, macrophages displayed a pro-inflammatory phenotype which facilitates embryo implantation. While, In the late firster trimester and second trimester, decidual macrophages are anti-proinflammatory which are helpful to pregnancy maintenance. Alterations in the ratio of pro-inflammatory/anti-inflammatory decidual macrophages lead to abortion, preeclampsia, and preterm birth. Placenta-derived exosomes (pEXO) are critical in the immune cell modulation such as T cell apoptosis, NK activities, and T regulatory (Treg) differentiation. However, it is unknown whether placenta-derived exosomes contribute to decidual macrophage polarization during early pregnancy. Here we report that exosomes from the placenta explant stimulate M2 macrophage polarization via exosomal miRNA-30d-5p. Mechanistically, miRNA-30d-5p polarized macrophages to M2 phenotype by inhibiting HDAC9 expression. Furthermore, the conditioned medium of pEXO-treated macrophages promoted trophoblast migration and invasion. By contrast, conditioned medium impaired the ability of endothelial cell tube formation. However, pEXO-treated macrophages have no impact on T cell proliferation. Together, we demonstrated that pEXO promoted trophoblast migration and invasion, endothelial cell migration, and attenuation of endothelial cell tube formation by polarizing macrophage to decidua-like macrophage.