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The microRNA profiles of extracellular vesicles (EVs) from osteoclasts (OCs) co-cultured with prostate cancer (PCa) cell lines [OCEVmiRNA-seq]

ABSTRACT: Bone metastatic lesions of PCa contain areas of bone destruction and formation that are directed by OCs and osteoblasts (OBs), respectively. OC-derived EVs are reported to maintain normal bone homeostasis; however, there is no study investigating the role of EVs from OCs in the presence of PCa.

ORGANISM(S): Mus musculus

PROVIDER: GSE268548 | GEO | 2024/06/06

REPOSITORIES: GEO

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The target gene investigation in osteoclats (OCs) treated with mmu-miR-5112 enriched in EVs from OCs with prostate cancer (PCa) cells [target_EV-miR-5112]

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Project description:Bone is a connective tissue which undergoes continuous remodeling, including bone resorption and formation by osteoclasts (OCs) and osteoblasts (OBs), respectively. Recently, extracellular vesicles (EVs) are increasingly appreciated as a regulator of intercellular communication between OCs and OBs. The zebrafish scale is a thin membranous bone embedded in the skin and consists of OBs, OCs, and bone matrix, providing an elegant model to visualize OC-OB communication. Here, we developed a double-transgenic zebrafish line, trap:GFP; osterix:mCherry, which expresses GFP and mCherry under the control of the OC-specific trap (tartrate-resistant acid phosphatase) and OB-specific osterix enhancer, respectively. Utilizing this double-transgenic line, in combination with Hoechst 33342 (Hoe) staining, we isolated four distinct fractions from the scale at 1 day post-fracture by flow cytometry, GFP(–) mCh(+) Hoe(high) (OB fraction), GFP(low) mCh(+) Hoe(high) (OC precursor (pOC) fraction), GFP(high) Hoe(high) (mature OC (mOC) fraction), and mCh(+) Hoe(–) (OB-derived extracellular vesicle (EV) fraction). In this study, we have performed RNA-seq analysis using these four fractions to examine the gene expression profile of each fraction.

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Project description:Young and Aged osteocytes in bone matrix secreted plenty of extracellular vesicles (EVs) with different functions. We found that EVs released from aged bone matrix (AB-EVs) during bone resorption favor adipogenesis rather than osteogenesis of BMSCs and augment calcification of vascular smooth muscle cells (VSMCs). In this work, we aim to detect the differential regulation microRNAs. Vascular calcification often occurs with osteoporosis, a contradictory association called “calcification paradox”. We find that extracellular vesicles (EVs) released from aged bone matrix (AB-EVs) during bone resorption favor adipogenesis rather than osteogenesis of BMSCs and augment calcification of vascular smooth muscle cells (VSMCs). Intravenous or intramedullary injection of AB-EVs promotes bone-fat imbalance and exacerbates Vitamin D3 (VD3)-induced vascular calcification in young or old mice. To explore the involvement of miRNAs in the AB-EVs-induced promotion of adipocyte formation and vascular calcification, the Agilent miRNA array was conducted to compare the miRNA expression profiles in AB-EVs and YB-EVs from mouse bone specimens. Our study uncovers the role of AB-EVs as a messenger for calcification paradox by transferring functional miRNAs.

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Bone density loci identified by genome-wide association studies segregate a lineage-specific PU.1-dependent gene regulatory network in osteoclasts [ChIP-seq]

Project description:Genome-wide association studies (GWASs) have been instrumental in understanding complex phenotypic traits. However, they have rarely been used to understand lineage-specific pathways and functions that contribute to the trait. In this study, by integrating publicly available lineage-specific enhancers from mesenchymal and myeloid compartments with bone mineral density loci, we were able to segregate osteoblast- and osteoclast (OC)-specific functions. Specifically, in OCs, a PU.1-dependent transcription factor (TF) network was revealed. Further we conducted functional genomic analysis PU.1 and MITF genome wide binding with corresponding gene expression analysis which revealed a TF network essential for OC differentiation. Several of these TFs were regulated by cooperative binding of PU.1 with BRD4 to form superenhancers. Further, PU.1 is essential for conformational changes in the superenhancer region of Nfatc1. In summary, our study demonstrates that utilizing GWASs with genome-wide binding studies as a filter helps to decipher lineage-specific pathways contributing to complex disease states.

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