Effect of culture conditions on microRNA expression in primary adult control and COPD lung fibroblasts in vitro

Ikari, Jun; Smith, Lynette M.; Nelson, Amy; Iwasawa, Shunichiro; Gunji, Yoko; Farid, Maha; Wang, Xingqi; Basma, Hesham; Feghali‐Bostwick, Carol; Liu, Xiangde; DeMeo, Dawn L.; Rennard, Stephen I.

doi:10.1007/s11626-014-9820-8

Cited by 16 publications

(8 citation statements)

References 48 publications

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“…In a previous study, we explored the effect of culture conditions on miRNA expression in primary adult lung fibroblasts from patients with and without COPD that included five subjects in each group [ 16 ]. In order to identify potentially differentially expressed miRNAs, we performed a volcano plot analysis of those data and identified miR-503 as an miRNA that consistently differed between COPD and control lung fibroblasts either in the absence or presence of IL-1ß and TNF-α ( Fig 1A and 1B ).…”

Section: Resultsmentioning

confidence: 99%

“…However, understanding of the precise role of specific miRNAs in defined cell types, including lung fibroblasts, is limited [ 10 ]. Formerly, we identified differentially expressed miRNAs in COPD lung fibroblasts in vitro [ 16 ]. Among the differentially expressed miRNA species, miR-503 expression was significantly reduced in cultured COPD lung fibroblasts under both basal and IL-1ß and TNF-α as assessed by microarray analysis.…”

Section: Discussionmentioning

confidence: 99%

“…Previously, by using miRNA-microarray analysis, we identified that expression of several miRNAs was consistently altered in COPD lung fibroblasts cultured in vitro and that this was independent of culture density or cell passage (between passages three and eight). Of these, miR-503 expression was decreased in COPD lung fibroblasts with or without IL-1β and TNF-α stimulation [ 16 ]. Although, miR-503 is abundantly expressed in the lung [ 17 ], the role of miR-503 and its target genes in lung fibroblasts has not been examined.…”

Section: Introductionmentioning

confidence: 99%

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Reduced microRNA-503 expression augments lung fibroblast VEGF production in chronic obstructive pulmonary disease

et al. 2017

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Alterations in microRNA (miRNA) expression may contribute to COPD pathogenesis. In COPD, lung fibroblast repair functions are altered in multiple ways, including extracellular mediator release. Our prior study revealed miR-503 expression is decreased in COPD lung fibroblasts, although the exact role played by miR-503 is undetermined. The current study examined a role of miR-503 in cytokine, growth factor and fibronectin production by lung fibroblasts from patients with and without COPD. Primary adult lung fibroblasts were isolated from patients with or without COPD. MiR-503 expression and interleukin (IL)-6, -8, PGE2, HGF, KGF, VEGF and fibronectin release were examined with or without inflammatory cytokines, IL-1β and tumor necrosis factor (TNF)-α. MiR-503 expression was decreased in COPD lung fibroblasts. The expression of miR-503 was positively correlated with %FVC, %FEV1, and %DLco as well as IL-6, -8, PGE2, HGF, KGF, and VEGF in the absence or presence of IL-1ß/TNF-α. In addition, IL-8 and VEGF release from COPD lung fibroblasts were increased compared to those from control. Exogenous miR-503 inhibited VEGF release from primary adult and fetal lung fibroblasts but not IL-8 release. As expected, COPD fibroblasts proliferated more slowly than control fibroblasts. MiR-503 did not affect proliferation of either control or COPD lung fibroblasts. MiR-503 inhibition of VEGF protein production and mRNA was mediated by direct binding to the 3’ untranslated region of VEGF mRNA. Endogenous miR-503 was differently regulated by exogenous stimulants associated with COPD pathogenesis, including IL-1ß/TNF-α, TGF-ß1 and PGE2. Endogenous miR-503 inhibition augmented VEGF release by IL-1ß/TNF-α and TGF-ß1 but not by PGE2, demonstrating selectivity of miR-503 regulation of VEGF. In conclusions, reduced miR-503 augments VEGF release from lung fibroblasts from patients with COPD. Since VEGF contributes to disturbed vasculature in COPD, altered miR-503 production might play a role in modulating fibroblast-mediated vascular homeostasis in COPD.

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Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Reduced microRNA-503 expression augments lung fibroblast VEGF production in chronic obstructive pulmonary disease

et al. 2017

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“…The overall similarity of genome-wide copy number and gene mutation patterns of paired CRCs and PBTs, was also observed in our cases at the miRNA expression level. Experimental studies have shown that both the biogenesis and expression levels of miRNAs are “susceptible” to effects of cell culture conditions, including the ones that affect cellular density and contact, replicated passages and senescence [ 55 – 59 ]. A recent study utilized miRNA expression to compare the effects of different cell reprogramming methods in cultured cells and medium [ 60 ].…”

Section: Discussionmentioning

confidence: 99%

Genomic comparison of early-passage conditionally reprogrammed breast cancer cells to their corresponding primary tumors

et al. 2017

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Conditionally reprogrammed cells (CRCs) are epithelial cells that are directly isolated from patients’ specimens and propagated in vitro with feeder cells and a Rho kinase inhibitor. A number of these cells have been generated from biopsies of breast cancer patients, including ductal carcinoma in situ and invasive carcinomas. The characterization of their genomic signatures is essential to determine their ability to reflect the natural biology of their tumors of origin. In this study, we performed the genomic characterization of six newly established invasive breast cancer CRC cultures in comparison to the original patients’ primary breast tumors (PBT) from which they derived. The CRCs and corresponding PBTs were simultaneously profiled by genome-wide array-CGH, targeted next generation sequencing and global miRNA expression to determine their molecular similarities in the patterns of copy number alterations (CNAs), gene mutations and miRNA expression levels, respectively. The CRCs’ epithelial cells content and ploidy levels were also evaluated by flow cytometry. A similar level of CNAs was observed in the pairs of CRCs/PBTs analyzed by array-CGH, with >95% of overlap for the most frequently affected cytobands. Consistently, targeted next generation sequencing analysis showed the retention of specific somatic variants in the CRCs as present in their original PBTs. Global miRNA profiling closely clustered the CRCs with their PBTs (Pearson Correlation, ANOVA paired test, P<0.05), indicating also similarity at the miRNA expression level; the retention of tumor-specific alterations in a subset of miRNAs in the CRCs was further confirmed by qRT-PCR. These data demonstrated that the human breast cancer CRCs of this study maintained at early passages the overall copy number, gene mutations and miRNA expression patterns of their original tumors. The further characterization of these cells by other molecular and cellular phenotypes at late cell passages, are required to further expand their use as a unique and representative ex-vivo tumor model for basic science and translational breast cancer studies.

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“…Similarly to asthma, a pathophysiological role for miRNAs in COPD has been suggested in several studies, in which differential expression of miRNAs has been identified in lung cell ( Figure 2 ) [ 41 , 42 ].…”

Section: Copdmentioning

confidence: 91%

Molecular Background of miRNA Role in Asthma and COPD: An Updated Insight

Szymczak

Wieczfińska

Pawliczak

2016

BioMed Research International

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Inflammatory airway diseases are a significant health problems requiring new approaches to the existing therapies and addressing fundamental issues. Difficulties in developing effective therapeutic strategies might be caused by lack of understanding of their exact molecular mechanism. MicroRNAs (miRNAs) are a class of regulators that already revolutionized the view of gene expression regulation. A cumulating number of investigations show a pivotal role of miRNAs in the pathogenesis of asthma, chronic obstructive pulmonary disease (COPD), or airway remodeling through the regulation of many pathways involved in their pathogenesis. Expression changes of several miRNAs have also been found to play a role in the development and/or improvement in asthma or COPD. Still, relatively little is known about the role of miRNAs in inflammatory disorders. The microRNA profiles may differ depending on the cell type or antigen-presenting cell. Based on the newest literature, this review discusses the current knowledge concerning miRNA contribution and influence on lung inflammation and chosen inflammatory airway diseases: asthma and COPD.

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Effect of culture conditions on microRNA expression in primary adult control and COPD lung fibroblasts in vitro

Cited by 16 publications

References 48 publications

Reduced microRNA-503 expression augments lung fibroblast VEGF production in chronic obstructive pulmonary disease

Reduced microRNA-503 expression augments lung fibroblast VEGF production in chronic obstructive pulmonary disease

Genomic comparison of early-passage conditionally reprogrammed breast cancer cells to their corresponding primary tumors

Molecular Background of miRNA Role in Asthma and COPD: An Updated Insight

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