MiRNA expression profile of human subcutaneous adipose and during adipocyte differentiation

Francisco J Ortega, José M Moreno-Navarrete, Gerard Pardo, Monica Sabater, Manuela Hummel, Anna Ferrer, Jose I Rodriguez-Hermosa, Bartomeu Ruiz, Wifredo Ricart, Belen Peral, José M Fernández-Real, Francisco J Ortega, José M Moreno-Navarrete, Gerard Pardo, Monica Sabater, Manuela Hummel, Anna Ferrer, Jose I Rodriguez-Hermosa, Bartomeu Ruiz, Wifredo Ricart, Belen Peral, José M Fernández-Real

Abstract

Background: Potential regulators of adipogenesis include microRNAs (miRNAs), small non-coding RNAs that have been recently shown related to adiposity and differentially expressed in fat depots. However, to date no study is available, to our knowledge, regarding miRNAs expression profile during human adipogenesis. Thereby, the aim of this study was to investigate whether miRNA pattern in human fat cells and subcutaneous adipose tissue is associated to obesity and co-morbidities and whether miRNA expression profile in adipocytes is linked to adipogenesis.

Methodology/principal findings: We performed a global miRNA expression microarray of 723 human and 76 viral mature miRNAs in human adipocytes during differentiation and in subcutaneous fat samples from non-obese (n = 6) and obese with (n = 9) and without (n = 13) Type-2 Diabetes Mellitus (DM-2) women. Changes in adipogenesis-related miRNAs were then validated by RT-PCR. Fifty of 799 miRNAs (6.2%) significantly differed between fat cells from lean and obese subjects. Seventy miRNAs (8.8%) were highly and significantly up or down-regulated in mature adipocytes as compared to pre-adipocytes. Otherwise, 17 of these 799 miRNAs (2.1%) were correlated with anthropometrical (BMI) and/or metabolic (fasting glucose and/or triglycerides) parameters. We identified 11 miRNAs (1.4%) significantly deregulated in subcutaneous fat from obese subjects with and without DM-2. Interestingly, most of these changes were associated with miRNAs also significantly deregulated during adipocyte differentiation.

Conclusions/significance: The remarkable inverse miRNA profile revealed for human pre-adipocytes and mature adipocytes hints at a closely crosstalk between miRNAs and adipogenesis. Such candidates may represent biomarkers and therapeutic targets for obesity and obesity-related complications.

Conflict of interest statement

Competing Interests: The authors have declared that no competing interests exist.

Figures

Figure 1. miRNA expression profiling during adipogenesis.
Figure 1. miRNA expression profiling during adipogenesis.
Scatter plot showing comparison of miRNA expression profiles between undifferentiated human pre-adipocytes (Day 0) and adipocytes during differentiation at 7th day (grey diamonds) or mature adipocytes at 14th day (black circles) after inducing differentiation of stem-cells from subcutaneous fat depots of both lean (Fig. 1A) or obese (Fig. 1B) subjects.
Figure 2. Specific miRNAs expression regulation during…
Figure 2. Specific miRNAs expression regulation during adipogenesis.
Expressions of miRNAs significantly deregulated during differentiation. MiRNAs expression levels are normalized to an internal control and plotted as fold changes in Log2-ratio scale for day 7 vs. day 0 (grey bars) and day 14 vs. day 0 (black bars). Log2-ratios of 2-class comparisons are symmetric around 0 (while a Log2-ratio of 1 means 2-fold up-regulation, a Log2-ratio of −1 means 2-fold down-regulation as compared to day 0 miRNAs levels). Data are expressed as mean ± SEM (n = 6). All the represented miRNAs (70 of the 799 analyzed miRNAs) showed ANOVA P-values <0.0001 for comparisons between groups and fold-changes over 1.2-fold or under −1.2-fold day 0 miRNA expression levels.
Figure 3. miRNAs expression profiling in human…
Figure 3. miRNAs expression profiling in human subcutaneous fat.
Scatter plot showing comparison of miRNA expression profiles between subcutaneous fat from non-obese (n = 6) and obese non-DM-2 (n = 13, grey diamonds) or obese and DM-2 (n = 9, black circles) women.
Figure 4. Human subcutaneous miRNAs expressions.
Figure 4. Human subcutaneous miRNAs expressions.
Expressions of miRNAs are normalized to an internal control and plotted as fold-changes in a Log2-scale for obese non-DM-2 vs. non-obese (Fig. A) or obese and DM-2 vs. non-obese (Fig. B). Data are expressed as mean ± SEM. The represented miRNAs showed in each case ANOVA P-values <0.05 for comparisons between groups and fold-changes over 1.2-fold or under −1.2-fold subcutaneous fat from non-obese miRNA level.

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