Developmental Programming: Prenatal Testosterone Excess on Liver and Muscle Coding and Noncoding RNA in Female Sheep

Abstract

Prenatal testosterone (T)-treated female sheep manifest peripheral insulin resistance, ectopic lipid accumulation and insulin signaling disruption in liver and muscle. This study investigated transcriptional changes and transcriptome signature of prenatal T excess-induced hepatic and muscle-specific metabolic disruptions. Genome-wide coding and non-coding (nc) RNA expression in liver and muscle from 21-month-old prenatal T-treated (T propionate 100mg intramuscular twice weekly from days 30 to 90 of gestation; Term: 147 days) and control females were compared. Prenatal T (1) induced differential expression of mRNAs in liver (15 down, 17 up) and muscle (66 down, 176 up) (FDR<0.05, absolute log2 fold change>0.5); (2) downregulated mitochondrial pathway genes in liver and muscle; (3) downregulated hepatic lipid catabolism and PPAR signaling gene pathways; (4) modulated ncRNA metabolic processes gene pathway in muscle and (5) downregulated 5 uncharacterized long ncRNA (lncRNA) in the muscle but no ncRNA changes in the liver. Correlation analysis showed downregulation of lncRNAs LOC114112974 and LOC105607806 was associated with decreased TPK1, and LOC114113790 with increased ZNF470 expression. Orthogonal Projections to Latent Structures Discriminant Analysis identified mRNAs HADHA and SLC25A45, and miRNAs MIR154A, MIR25 and MIR487B in liver and ARIH1 and ITCH and miRNAs MIR369, MIR10A and MIR10B in muscle as potential biomarkers of prenatal T-excess. These findings suggest downregulation of mitochondria, lipid catabolism, and PPAR signaling genes in liver and dysregulation of mitochondrial and ncRNA gene pathways in muscle are contributors of lipotoxic and insulin resistant hepatic and muscle phenotype. Gestational T excess programming of metabolic dysfunctions involve tissue-specific ncRNA modulated transcriptional changes.

Genes in loci genetically associated with polycystic ovary syndrome are dynamically expressed in human fetal gonadal, metabolic and brain tissues

“…Notably, brothers/sons of women with PCOS have elevated androgen levels ( 53 ), increased total cholesterol and low density lipoproteins levels at puberty ( 16 ), decreased insulin sensitivity (independent of obesity) and glucose tolerance ( 12 ), among other symptoms. Also, hepatic dysfunctions and risk of liver diseases have been observed in PCOS models in male sheep ( 11 ), female sheep ( 54 , 55 ), and rats ( 56 , 57 ), together affirming the roles of these organs in the pathophysiology of the syndrome. Although transcriptional and post-transcriptional factors were not evaluated, INSR and FDFT1 were dynamically expressed in all tissues examined in this study.…”

Developmental programming: adverse sexually dimorphic transcriptional programming of gestational testosterone excess in cardiac left ventricle of fetal sheep

“…In our current study we utilized complementary analytical approaches namely differential expression analysis, traditionally used and dimension reduction multivariate modeling (PCA and PLS-DA) analysis. Such complementary approaches have been recently used in our prenatal T excess model assessing coding and non-coding transcriptome alteration in the liver and muscle 60 . Multivariate dimensionality reduction modeling condense related information into fewer weighted variables reducing model complexity and retaining prediction power compared to univariate analysis.…”

“…These techniques are useful in analyzing coding and non-coding RNA data simultaneously as many of the ncRNA are expressed in very low levels and with high number of zero values in the data which may result in reduced power to detect significance in the traditional methods using negative binomial distribution (e.g. differential expression analysis) 60 . Higher Variable Importance in Projection (VIP) value as listed in the tables indicate higher importance to the model and variables responsible for the separation of the two groups in the PLS DA model, some of these potential signatures also met the FDR adjusted significance criteria as indicated in the “ Results ” section.…”

Nutritional programming of white adipose tissue and mitochondrial function : Fat quality matters

“…Mitochondrial functionality is essential for the regulation of the hepatic lipid metabolism, but excessive lipid accumulation can impair hepatic mitochondrial function, as shown in a mice model for obesity (83). Indeed, hepatic mitochondrial function was altered in T2D and NAFLD, suggested to be affected by excessive lipid storage in the liver (84,85). In particular, hepatic accumulation of intermediates of triglyceride synthesis, like diacylglycerols, and changes in specific phospholipid species, like ceramides, are associated with adverse metabolic consequences (86,87).…”

“…Our research contributes to the previous findings of others describing that mitochondrial function can be programmed by (dietary) factors in prenatal, perinatal, and postnatal life (Figure 2). Currently, most evidence is from studies evaluating the impact of Additionally, evidence exists for the programming of offspring mitochondrial function 7 by maternal health status, like obesity ( 83) and (gestational) diabetes (16,24,83) and maternal hormones levels, like testosterone (84,85), glucocorticoids (86) and the stress hormone cortisol (87) as well as maternal stress (88) (Figure 2 and Supplementary Table 1 for an overview). The number of studies investigating the sole effect of the early postnatal diet on mitochondrial function are, however, limited.…”