The pro-fibrotic environment in muscle wasting [13], CBA increased the expression ofThe pro-fibrotic environment in

The pro-fibrotic environment in muscle wasting [13], CBA increased the expression of
The pro-fibrotic environment in muscle wasting [13], CBA increased the expression of the gene encoding the transmembrane protein 119 (TMEM119), which drives the differentiation of myoblasts into osteoclasts, a cell type that promotes the deposition of collagen. CBA altered expression of genes important for neuronal survival and regeneration, CNS development, acetylcholine receptor expression, and neuronal organization and orientation, consistent with a CBA-mediated alteration of NMJ and normal muscle function.Muscle satellite cell growth and survival categoryConsistent with our previous findings that CBA impairs skeletal muscle regenerative capacity, the PD98059 price results from this study showed that CBA altered the expression of 38 genes that contribute to the growth, survival, and cell cycle regulation of PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/28388412 cells. These include genes important for cell cycle progression, growth factor related signaling pathways, cellular survival and apoptosis, and additional genesWe identified 35 miRNAs whose expression changed 1.5-fold in a CBA-dependent manner. Of these, 14 were downregulated and 21 were upregulated (Fig. 2a). Functional enrichment of predicted target genes of differentially expressed miRNAs was analyzed using miRsystem as described in the methods section. Pathways associated with MAPK signaling, insulin signaling, neuronal signaling, and focal adhesions were enriched (Fig. 2b). The 100 most enriched pathways that included 20 differentially expressed miRNAs, 20 target genes, and had a score 1 are shown in Table 1. Target genes from additional pathways showed a higher representation of proteins important for essential enzymes or cofactors in ubiquitin-mediated degradation of proteins, PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/29072704 genes involved in maintenance of the integrity or in the breakdown of the extracellular matrix and muscle contraction (Additional file 3: Table S3). Expression values for 3 upregulated candidate miRNAs (miR-34a, miR-10b and miR-20) in the microarray were confirmed by qPCR (Fig. 2c). We observed significant CBA/SIV-dependent changes in miRNA gene expression consistent with our microarray results. We also determined expression levels of 3 mRNAs: (1) estrogen receptor-alpha (ESR1), which demonstrated CBA/SIVdependent changes in the microarray results and serves as a validated target of miR-34a and -20, (2) B-cell lymphoma-2 (BCl-2), a validated target of miR-34a, and (3) Kirsten Rat Sarcoma (KRAS), a validated target of miR-18. We found significant changes in ESR1 that were consistent with our microarray, and all mRNA showed significant changes with the expected inverse correlation to miRNA changes (Fig. 2c). As described in the methods, we also analyzed biological functions of target genes of differentially regulated miRNAs. Genes targeted by downregulated miRNAs are involved in inflammatory/immune response, general cellular functions, neuronal function, etc. Frequently, a single gene was targeted by a miRNA within each individual biological function. Nearly 50 of the genes targeted by upregulated miRNAs are primarily associated with five biological functional groups including (1) regulation of cell cycle progression, (2) activation of the innate immune response upon viral infection, (3) control of neuronal survival and plasticity, CNS development, and neuronal differentiation, (4) multifunctional receptors that mediate several cellular functions (e.g., proliferation, differentiation, glucoseSimon et al. BMC Genomics (2015) 16:Page 5 ofFig. 2 Functional enrichment.