miRNA expression in the failing human heart: functional correlates - PubMed (original) (raw)

miRNA expression in the failing human heart: functional correlates

Carmen Sucharov et al. J Mol Cell Cardiol. 2008 Aug.

Abstract

MicroRNAs (miRNAs) are small, noncoding ~22-nucleotide regulatory RNAs that are key regulators of gene expression programs. Their role in the context of the cardiovascular system has only recently begun to be explored; however, changes in the expression of miRNAs have been associated with cardiac development and with several pathophysiological states including myocardial hypertrophy and heart failure. We demonstrate that miRNA expression patterns are distinct in two types of heart failure: idiopathic dilated cardiomyopathy and ischemic cardiomyopathy. To pursue the observation that changes in expression levels of individual miRNAs are functionally relevant, microRNA mimics and inhibitors to miR-92, miR-100 and miR-133b were expressed in primary cultures of neonatal rat cardiac myocytes. These studies demonstrated that over-expression of miR-100 is involved in the beta-adrenergic receptor-mediated repression of "adult" cardiac genes (i.e., alpha-myosin heavy chain, SERCA2a), and that over-expression of miR-133b prevents changes in gene expression patterns mediated by beta-adrenergic receptor stimulation. In conclusion, some miRNA expression patterns appear to be unique to the etiology of cardiomyopathy and changes in the expression level of miRs 100 and 133b contribute to regulation of the fetal gene program. It is likely that this miR-directed reprogramming of key remodeling genes is involved in the establishment and progression of common human cardiomyopathies.

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Figures

Figure 1

Figure 1

miRNA expression profiles in samples obtained from non-failing (NF), idiopathic cardiomyopathy (IDC) and ischemic (ISC) patients. (A) Relative expression of miRNAs is expressed at the log base 2 ratio of failing (F)/nonfailing (NF). NF vs. IDC (red), NF vs. ISC (blue). Only miRNAs with a p-value < 0.10 as determined by t-Test are shown. (B) Using miRNA specific ABI primers, the relative expression of a subset of miRNAs was confirmed by RT-PCR, as described in Methods.

Figure 1

Figure 1

miRNA expression profiles in samples obtained from non-failing (NF), idiopathic cardiomyopathy (IDC) and ischemic (ISC) patients. (A) Relative expression of miRNAs is expressed at the log base 2 ratio of failing (F)/nonfailing (NF). NF vs. IDC (red), NF vs. ISC (blue). Only miRNAs with a p-value < 0.10 as determined by t-Test are shown. (B) Using miRNA specific ABI primers, the relative expression of a subset of miRNAs was confirmed by RT-PCR, as described in Methods.

Figure 2

Figure 2

Over-expression or down-regulation of a subset of miRNAs in NRVMs regulates β-AR changes in gene expression. (A) Changes in miR-92, miR-133b and miR-100 abundance upon transfection with microRNA mimics or microRNA inhibitors. miRNA abundance was determined by RT-PCR. (B-D) Changes in fetal gene expression in NRVMs transfected with microRNA mimic or microRNA inhibitors. Cells were treated with 10-7M ISO (black bars) 24 hours after transfection and harvested 48 hours after treatment. Gene expression was measured by RT-PCR. Results were normalized to 18S rRNA expression levels and were compared to cells transfected with a control mimic or inhibitor, defined as 1 (line at 1). The graphs represent an average of 3-4 individual experiments.

Figure 2

Figure 2

Over-expression or down-regulation of a subset of miRNAs in NRVMs regulates β-AR changes in gene expression. (A) Changes in miR-92, miR-133b and miR-100 abundance upon transfection with microRNA mimics or microRNA inhibitors. miRNA abundance was determined by RT-PCR. (B-D) Changes in fetal gene expression in NRVMs transfected with microRNA mimic or microRNA inhibitors. Cells were treated with 10-7M ISO (black bars) 24 hours after transfection and harvested 48 hours after treatment. Gene expression was measured by RT-PCR. Results were normalized to 18S rRNA expression levels and were compared to cells transfected with a control mimic or inhibitor, defined as 1 (line at 1). The graphs represent an average of 3-4 individual experiments.

Figure 2

Figure 2

Over-expression or down-regulation of a subset of miRNAs in NRVMs regulates β-AR changes in gene expression. (A) Changes in miR-92, miR-133b and miR-100 abundance upon transfection with microRNA mimics or microRNA inhibitors. miRNA abundance was determined by RT-PCR. (B-D) Changes in fetal gene expression in NRVMs transfected with microRNA mimic or microRNA inhibitors. Cells were treated with 10-7M ISO (black bars) 24 hours after transfection and harvested 48 hours after treatment. Gene expression was measured by RT-PCR. Results were normalized to 18S rRNA expression levels and were compared to cells transfected with a control mimic or inhibitor, defined as 1 (line at 1). The graphs represent an average of 3-4 individual experiments.

Figure 2

Figure 2

Over-expression or down-regulation of a subset of miRNAs in NRVMs regulates β-AR changes in gene expression. (A) Changes in miR-92, miR-133b and miR-100 abundance upon transfection with microRNA mimics or microRNA inhibitors. miRNA abundance was determined by RT-PCR. (B-D) Changes in fetal gene expression in NRVMs transfected with microRNA mimic or microRNA inhibitors. Cells were treated with 10-7M ISO (black bars) 24 hours after transfection and harvested 48 hours after treatment. Gene expression was measured by RT-PCR. Results were normalized to 18S rRNA expression levels and were compared to cells transfected with a control mimic or inhibitor, defined as 1 (line at 1). The graphs represent an average of 3-4 individual experiments.

Figure 3

Figure 3

Over-expression or down-regulation of miRNA-133b in NRVMs regulates cellular hypertrophy. Cells were transfected with miR-133b mimic or inhibitors and cell surface area was quantified. (A) Immunofluorescence with anti-actinin antibody. (B) Cell size was measured using the Image J software. A total of 30 cells from 3 different fields were measured.

Figure 3

Figure 3

Over-expression or down-regulation of miRNA-133b in NRVMs regulates cellular hypertrophy. Cells were transfected with miR-133b mimic or inhibitors and cell surface area was quantified. (A) Immunofluorescence with anti-actinin antibody. (B) Cell size was measured using the Image J software. A total of 30 cells from 3 different fields were measured.

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