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MicroRNA global profiling in cystic fibrosis cell lines reveals dysregulated pathways related with oncogenesis, inflammation, growth and glucose metabolism

ABSTRACT: Global profiles of 754 human miRNAs from CFBE41o- (F508del/F508del), and IB3 (F508del/W1282X) cell lines, carrying the most common mutations for Cystic Fibrosis, were analysed. 16HBE14o- cells, derived from normal bronchus, were used as calibrator. The relative abundance of each miRNA was calculated and fold-changes≥+2 or ≤-2 (p-value≤0.05) were considered significant. MiRNA target genes and KEGG pathways were identified using miRWalk v.3 and DIANA-mirPath v3.0, respectively and a protein-protein interaction network was performed by STRING 11.0. In CF cell lines 41 miRNAs showed significant changes. Eight miRNAs were regulated in CFBE41o- only and 23 in IB3 only, suggesting genotype-specific effects. Five miRNAs were up-regulated and 1 down-regulated in both CFBE41o- and IB3. The 41 miRNAs regulated genes within pathways involved with cancer, inflammation, body growth, glucose metabolism and lipid metabolism. Interestingly, key genes involved with Ras, TGF-beta, Jak-Stat and insulin signaling were highlighted.

ORGANISM(S): Homo sapiens

PROVIDER: GSE167100 | GEO | 2022/02/28

REPOSITORIES: GEO

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Project description:Purpose: The aim of present research was to identify miRNAs potentially related with fattness traits in pigs Methods: miRNA-seq analysis was performed on subcutaneous fat samples collected from 22 pigs representing two sire-line breeds – Pietrain and Hampshire and one dam-line – Large White. Based on dissection data, pigs were selected from a larger population in terms of fatness traits to obtain the most extreme groups in each breed. The cDNA libraries were constructed form 300 ng of total RNA with the use NEBNext Multiplex Small RNA Library Prep Set for Illumina (New England Biolabs, Ipswich, MA, United States) according to the protocol. The quantification of obtained libraries was performed on Qubit 2.0 (Invitrogen, Life Technologies) and TapeStation 2200 (D1000 ScreenTape; Agilent). The RNA-seq was performed in 36 single-end cycles on HiScanSQ platform (Illumina) with the use of ruSeq SR Cluster Kit v3- CBOT-HS and TruSeq SBS Kit v 3 - HS (Illumina). Each library was sequences in 4 technical replications. The DEGs were detected using DESEq2 software and validation was performed by qPCR. Results: RNA-seq approach allowed to identify miRNAs differentially expressed between pigs with various fatness traits in each analyzed breeds: 64 for Pietrain pigs; 41 for Hampshire and 46 for Large White pigs (pvalue <0.05; fold change≤1.5). The comparison of obtained miRNAs sets showed 13 miRNAs identified in all three breeds. The Gene Ontology analysis (mirPath v.3.0 DIANA Tools web with DIANA—TarBase v7.0 as a reference) confirmed that detected miRNAs were involved in fatty acid biosynthesis (pvalue <1e-325); fatty acid metabolism (pvalue 6.661338e-16); ECM-receptor interaction (pvalue <1e-325). The most important targeted genes regulated by identified miRNAs were: fatty acid synthase – FASN; Acetyl-CoA carboxylase 1 – ACACA; Malonyl CoA-acyl carrier protein transacylase – MCAT and acetyl-CoA acyltransferase 1 and 2 (ACAA1 and ACAA2 genes). Conclusions: Obtained results allow to propose the panel of miRNAs, which can be related with fatness in pig. Such analysis can be the basis of future research in terms of identification of molecular mechanisms related with adipogenesis and fatness traits in pigs.

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MicroRNA global profiling in cystic fibrosis cell lines reveals dysregulated pathways related with oncogenesis, inflammation, growth and glucose metabolism

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