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Transcriptome sequencing in serum exosomes from proliferative diabetic retinopathy (PDR)

ABSTRACT: Purpose: To detect serum exosomal ncRNA profiles of proliferative diabetic retinopathy (PDR) by High-throughput sequencing Methods: serum exosomal non-coding RNA (ncRNA) profiles profiles of PDR and MH were generated by deep sequencing, only in once, using IlluminaHiSeq 3000. After analyzing the base composition and quality of the data, according to the analysis results of the original data, the data were filtered to remove the joint sequence and the contaminated part, and to remove low-quality base sequences.If it is paired-ended sequencing data, the filtered data should be further screened to retain the paired sequences and obtain clean data. Conclusions: Our study represents the first detailed analysis of serum exosomal transcriptomes from PDR patients by RNA-seq technology. The optimized data analysis workflows reported here should provide a framework for comparative investigations of expression profiles.

ORGANISM(S): Homo sapiens

PROVIDER: GSE178721 | GEO | 2021/06/24

REPOSITORIES: GEO

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Project description:Clinical management of metastatic gastric cancer (mGC) remains a major challenge due to a lack of specific biomarkers and effective therapeutic targets. Recently, accumulating evidence has suggested that exosomes play an essential role in cancer metastasis and can be an excellent reservoir of novel biomarkers and candidate therapeutic targets for cancer. Therefore, in this study, we aimed to reveal the proteomic profile of mGC-derived exosomes.Exosomes were isolated from pooled serum samples of 20 mGC patients and 40 healthy controls (HCs) by ultracentrifugation. Next, quantitative proteomic analyses were applied to analyze the protein profiles of the exosomes, and bioinformatic analyses were conducted on the proteomic data. Finally, the expression of exosomal protein candidates was selectively validated in individual subjects by western blot analysis. We isolated exosomes from serum samples. The size of the serum derived exosomes ranged from 30 to 150 nm in diameter. The exosomal markers CD9 and CD81 were observed in the serum exosomes. However, the exosomal negative marker calnexin, an endoplasmic reticulum protein, was not detected in exosomes. Overall, 443 exosomal proteins, including 110 differentially expressed proteins (DEPs) were identified by quantitative proteomics analyses. The bioinformatics analyses indicated that the upregulated proteins were enriched in the process of protein metabolic, whereas the downregulated proteins were largely involved in cell-cell adhesion organization. Surprisingly, ten highly vital proteins (UBA52, PSMA1, PSMA5, PSMB6, PSMA7, PSMA4, PSMA3, PSMB1, PSMA6, and FGA) were filtered from DEPs, most of which are proteasome subunits. Moreover, the validation data confirmed that PSMA3 and PSMA6 was explicitly enriched in the serum derived exosomes from patients with mGC. The present study provided a comprehensive description of the serum exosome proteome of mGC patients, which could be an excellent resource for further studies of mGC.

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Transcriptome sequencing in serum exosomes from proliferative diabetic retinopathy (PDR)

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