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Human osteoclast derived from SAPHO PBMC

ABSTRACT: Inducing the PBMCs from SAPHO patients differentiation into osteoclasts while stimulating with serum exosomes. Compared with exosomes from normal volunteer, we found that about 2/3 of patients’ osteoclast differentiation were inhibited in response to exosomes from SAPHO patients. We present a TMT-labeled quantitative proteomics analysis of the osteoclasts which respond differently to different exosomes.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Cell Culture

SUBMITTER: Wei Ge

LAB HEAD: Wei Ge

PROVIDER: PXD012218 | Pride | 2021-09-08

REPOSITORIES: Pride

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			Action	DRS
	MS181501SAPHO-PBMC.msf	Msf
	MS181501SAPHO-PBMC.pdResult	Other
	MS181501SAPHO-PBMC.pep.xml	Pepxml
	MS181501SAPHO-PBMC.xlsx	Xlsx
	MS181501_1.raw	Raw

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Publications

Serum-derived extracellular vesicles inhibit osteoclastogenesis in active-phase patients with SAPHO syndrome.

Gao Yanpan Y Chen Yanyu Y Wang Lun L Li Chen C Ge Wei W

Therapeutic advances in musculoskeletal disease 20210416

<h4>Objective</h4>Synovitis, acne, pustulosis, hyperostosis, and osteitis (SAPHO) syndrome is a rare chronic inflammatory disorder and the underlying pathogenesis is unclear. In this study, 88 SAPHO patients and 118 healthy controls were recruited to investigate the role of serum-derived extracellular vesicles (SEVs) in SAPHO syndrome.<h4>Methods</h4>Quantitative proteomics was applied for SEVs proteome identification, and ELISA and Western blotting was performed to verify the results of mass sp ...[more]

PMID: 33948126

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Quantitative proteomics profiling revealed serum exosomes inhibited the differentiation of osteoclast in SAPHO patients during the active period

Project description:The serum samples from patients and normal volunteers were pooled together, respectively, to perform isolation of exosomes by ultracentrifugation.TMT-labeled quantitative proteomics were performed to find out the differently expressed proteins.

2021-09-08 | PXD012125 | Pride

Transcriptome-wide identification of coding and noncoding RNA-binding proteins defines the extended RNA interactome of Leishmania mexicana

Project description:Non-poly(A) RNA molecules including noncoding RNAs (ncRNAs) comprise the major portion of the total transcribed molecules in the cell. In addition to the mRNAs the ncRNAs also function as ribonucleoprotein particles (RNPs) and carry out biological functions including synthesis of new proteins, RNA processing, genome remodelling and regulation of transcription. We therefore envisaged a comprehensive transcriptome-wide identification of coding and non-coding RNA-binding proteins (RBPs) in the Leishmania spp. Towards this we applied the recently reported orthogonal organic phase separation (OOPS) method in combination with tandem mass tag (TMT) labelling-based quantitative proteomic mass spectrometry (MS) and report herein the most comprehensive identification of RBPs in Leishmania mexicana (L. mexicana) parasites. This study identified novel RNA binding property of thousands of L. mexicana proteins, significantly expanding the RBP landscape of the parasite. Furthermore, we showed that the classical Hsp90 inhibitor tanespimycin differentially regulates the RNA-binding property of hundreds of L. mexicana RBPs, shedding light into hitherto unknown large-scale downstream molecular effects of the small molecule inhibitor in the parasite.

2022-01-14 | PXD028092 | Pride

Involvement of serum-derived exosomes of patients with low bone density in progressive failure of bone remodeling via alteration of intercellular signal transduction and integrin-mediated mechanosensation

Project description:Exosomes are secreted into the blood by various types of cells. These small vesicles are involved in bone remodeling by mediation of intercellular communication in osteoblasts, osteoclasts or their precursors. Alterations in exosomal proteins are related to the failure of bone remodeling, which results in progressive loss of bone mass and poor quality of bone structure. However, the molecular changes in serum-derived exosomes (SDEs) from patients with low bone density and their functions in bone remodeling remain to be fully elucidated. We present a quantitative proteomics analysis of exosomes purified from the serum of patients with osteoporosis/osteopenia and normal volunteers; these data are available via Proteome Xchange with the identifier PXDXXXXX. Overall, 1,371 proteins were identified with an overlap of 1,160 Gene IDs among the ExoCarta proteins. In vitro studies and bioinformatics analysis revealed that the main changes in the SDEs of osteoporosis patients were proteins playing critical roles in integrins-mediated mechanosensation and signaling cascades, which are implicated in aggravating the failure of bone remodeling, including the enhancement of osteoclast differentiation and inhibition of bone mineralization. In contrast, the main changes in SDEs of osteopenia patients were proteins known to facilitate both osteoclast differentiation and osteoblastic bone formation, which resulted in a compensatory elevation of bone remodeling. In addition, bioinformatics analysis indicated that SDEs from elderly volunteers mediate negative regulation of bone remodeling via selenium deficiency-associated oxidative stress. This information will be helpful in elucidating the pathophysiological functions of SDEs and aid in the development of osteoporosis diagnostics and therapeutics.

2018-04-09 | PXD006463 | Pride

Inhibition of DOT1L increases osteoclast fusion and resorption and aggravates bone loss

Project description:Osteoclasts are absorptive cells and play a critical role in homeostatic bone remodeling and pathological bone resorption. Emerging evidence suggests an important role for epigenetic regulation of osteoclastogenesis. In this study, we investigated the role of DOT1L, which regulates gene expression epigenetically by histone H3K79 methylation during osteoclast formation. DOT1L and H3K79me2 levels were upregulated during osteoclast differentiation. Small molecule inhibitor- (EPZ5676 or EPZ004777) or short hairpin RNA-mediated reduction in DOT1L expression promoted osteoclast differentiation and resorption. DOT1L inhibition also increased osteoclast area and accelerated bone mass reduction in a mouse ovariectomy (OVX) model of osteoporosis. DOT1L inhibitors did not alter osteoblast differentiation in vitro and in vivo. Proteomics data, together with bioinformatics analysis, revealed that DOT1L inhibition altered reactive oxygen species (ROS) generation, autophagy activation, and cell fusion-related protein expression. ROS generation increased, and autophagy activation and cell migration ability enhancement were verified subsequently by flow cytometry and transwell migration assays. DOT1L inhibition increased NFATc1 nuclear translocation and NF-κB activation and strengthend osteoclast fusion and expression of resorption-related protein CD9, and MMP9 in osteoclasts derived from RAW264.7. Our findings support a new mechanism of DOT1L-mediated H3K79me2 epigenetic regulation of osteoclast differentiation, implicating DOT1L as a new therapeutic target for osteoclast dysregulation-induced disease.

2018-01-24 | PXD006448 | Pride

Quantitative proteomics analysis of exosomes purified from serum of patients with colorectal cancer

Project description:Tumor cells of colorectal cancer (CRC), release exosomes into peripheral blood. These exosomes can mediate communication between cells and affect various tumor-related processes in their target cells. We present a quantitative proteomics analysis on the exosomes purified from serum of patients with CRC and normal volunteers. 918 proteins were identified with an overlap of 725 Gene IDs in Exocarta proteins list. Comparing with the serum-purified exosomes of normal volunteers, we found 36 proteins upregulated and 22 proteins downregulated in the serum-purified exosomes of CRC patients, of which over 50% were involved in protein metabolism and immune response. Pathway and interaction network analysis results revealed that most of the exosome-enriched differentially expressed proteins (DEPs) were associated with complement and coagulation cascades and cancer pathways, suggesting their potential roles in tumor carcinogenesis. Our study demonstrates that serum-purified exosomes of CRC patients can play a pivotal role in invasiveness and pre-metastatic niche establishment of CRC, but not sustain the tumor survival and proliferation. The content selection of exosomes might be associated with the functions of proteins in the development of CRC. This information will be helpful in elucidating the pathophysiological functions of tumor-derived exosomes, and aid in the development of colorectal cancer diagnostics and therapeutics.

2016-12-12 | PXD003875 | Pride

serum exosomes contain SPARC and LRG1 as biomarkers for colon cancer and differ by tumor primary location

Project description:Purpose: Over the past few years, the distinction between left- and right-sided colon cancer has been brought into focus. Right-sided tumor location was associated with an inferior overall survival and progression-free survival. We aimed to perform a detailed analysis of the diversity in exosomes between left- and right-sided colon cancer using quantitative proteomics. Experimental Design: We isolated exosomes from left- and right-sided colon cancer patients and healthy volunteers and treated colorectal cancer cell line with serum-derived exosomes. Then we performed quantitative proteomics analysis of the serum-derived exosomes and cell line treated with exosomes, respectively. Results: The expression profile of the serum exosome proteome in patients with right-sided colon cancer is different from patients with left-sided colon cancer. Serum-derived exosomes of right-sided colon cancer promote metastasis via up-regulation of extracellular matrix-related proteins, especially proteoglycans like SPARC and glycoprotein like LRG1. Exosomal SPARC and LRG1 were closely correlated with progression-free survival. Conclusions: Proteomic analysis identified different exosomal protein profiling between left- and right-sided colon cancer. Serum-derived exosomes of right-sided colon cancer promote metastasis via overexpression of SPARC and LRG1.

2019-11-25 | PXD012283 | Pride

serum exosomes of right-sided colon cancer promote metastasis via overexpression of SPARC and LRG1

2019-11-25 | PXD012304 | Pride

Osteoclast-derived exosomal miR-5134-5p interferes with alveolar bone homeostasis by targeting the JAK2/STAT3 axis

Project description:The specific mirnas in the exosomes of mature osteoclasts were found by exosome sequencing, and their functions were identified by acting on osteoblasts to observe the differentiation and osteogenesis of osteoblasts

2023-01-04 | GSE222004 | GEO

Plasma exosomal proteomic pattern in Hemophagocyticlymphohistiocytosis of a twin boy

Project description:Background: Secondary HLH is closely related to EBV infection, protein changes in plasma exosomes may be related to the pathogenesis of HLH patient. In recent years quantitative proteomic has been used to clarify the relationship between basic medicine and clinical medicine. Methods: Exosomes were isolated from plasma samples of a HLH patient and his twin brother. Plasma samples were classified into three groups according to the clinical presentation, namely T1 (the acute HLH), T2 (during pharmacological treatments), Ctr (the healthy control). Herein, we used TMT-based LC-MS/MS platform to identified and characterized HLH signatures before and during pharmacological treatments. Results: Exosomes’ marker proteins- CD63, TSG101, HSP70 were identified by western blotting. Through GO analysis and KEGG analysis, we found that the signaling pathways and functions enriched by differentially expressed genes are very different. We focus on describing? the significantly up-regulated proteins-MSN, HSPA8, CD163, and the significantly down-regulated proteins- PLG, FGG, F2. The significant changes of these factors are related to the deterioration of the disease Conclusion: Our exploratory study shows that the clinical symptoms and pathophysiological changes of HLH driven by EBV might be reflected in patients’ plasma exosomes. Plasma exosome proteomics provides new highlightinsights for into exploring the diagnostic and therapeutic biomarkers of HLH.

2022-05-19 | PXD019088 | Pride

Global phosphoproteome of Leishmania mexicana developmental stages

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