Transcriptome analyses reveal metabolic syndrome of adulthood with intrauterine growth restriction in a pig model
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ABSTRACT: Epidemiological data indicated intrauterine growth retardation (IUGR) is a risk factors for adult metabolic syndrome. However, the genetic mechanism underlying the phenotype in adulthood is not well characterized. Here, the present study employed adult normal and IUGR pigs as models to survey the difference of global gene expression in the liver using transcriptome sequence. The trancriptome libraries generated 104.54 gigabases data, 16,948 and 17,078 genes were expressed in normal and IUGR pigs, repectively. A total of 1,322 differentially expreesion genes (DGEs) were identified. An enrichment analysis of DGEs presented the top overrepresented GO terms and pathway were related to oxidoreductase activity, ATPase activity, amino catabolic process, glucose metabolism and Insulin signaling pathway. Finally, combined their phenotypes and gene expression patterns, we proved the adult IUGR pigs would obstruct the process of mitochondrial biogenesis and oxidative phosphorylation, which induced high oxidation press in adult IUGR. The increased gluconeogenic and decreased glycogen synthesis in liver resulted in reducing the capacity of glucose intolerance in IUGR. The decreased expression of insulin signaling genes (such as PI3K and AKT) implied a risk of diabetes in adult IUGR. Moreover, the capacity of fatty acid oxidation decreased in IUGR liver leaded to high content of triglyceride and free fatty acid in IUGR. Together, these findings provide a comprehensive understanding of the molecular mechanisms of adult IUGR pigs and valuable information for future studies of therapeutic intervention in IUGR metabolic syndrome.
Project description:Physiologically the liver is exposed to higher insulin concentrations than other organs. To evaluate functional consequences of insulin-deficient diabetes mellitus for the liver, we used a genetically engineered pig model of mutant INS gene induced diabetes of youth (MIDY). Liver samples of MIDY pigs and wild-type (WT) littermate controls were analyzed by RNA sequencing, label-free proteomics and targeted metabolomics/lipidomics to reveal pathways and key drivers significantly affected by chronic insulin deficiency and hyperglycemia. Gene set enrichment analysis of the ~500 transcripts that were differently abundant between MIDY and WT samples revealed pathways related to amino acid metabolism, beta-oxidation of fatty acids, gluconeogenesis and ketogenesis to be enriched in MIDY samples, whereas pathways related to extra cellular matrix and inflammation/pathogen defense response were enriched in the WT samples. Reduced insulin receptor activation and phosphorylation of protein kinase B (PKB, AKT) was associated with markedly increased levels of retinol dehydrogenase 16 (RDH16) and 3-hydroxy-3-methylglutaryl-CoA synthase (HMGCS2), the apparent key drivers of stimulated gluconeogenesis and ketogenesis in MIDY pigs. Profiling of acylcarnitines provided evidence for increased activity of carnitine palmitoyltransferase 1 (CPT1) shuttling fatty acids into the mitochondrial matrix for beta-oxidation and for increased omega-oxidation. In addition, several enzymes involved in amino acid degradation and enzymes of the urea cycle were increased in abundance, consistent with an increased use of amino acids for gluconeogenesis. Transcripts and proteins related to extracellular matrix, such as collagens, and inflammatory/immune mechanisms, such as C-reactive protein, proteins involved in or regulated by Toll-like receptor signaling and components of major histocompatibility complexes, were less abundant in MIDY vs. WT liver samples. Our study provides the first multi-omics analysis of liver in a clinically relevant large animal model for insulin-deficient diabetes mellitus.
Project description:Intrauterine growth restriction (IUGR) is associated with increased relative liver weight at birth, hepatic function decline, and a higher risk for chronic liver and cardiovascular diseases in adults. Precise mechanisms of early developmental plasticity to intervene in poor fetal programming and adult disease remain largely elusive and warrant extensive research. Selecting natural piglets’ model of IUGR, using the liver as a readout and combining previous transcriptome findings, a map of cellular landscape was created to reveal a sex-dependent manner in IUGR-induced hepatic injury and its long-term functional repercussions.Here, we show data on the transcriptional profiles of 41,969 high-quality cells from normal birthweight (NBWs) and IUGR piglets (IUGRs) from hepatic tissue and demonstrated strong homology with human using human-derived liver single-cell dataset. We discovered that male liver was much more severely damaged and inflammation by IUGR than female liver at the one-week postnatal node.
Project description:Intrauterine growth restriction (IUGR) is associated with increased relative liver weight at birth, hepatic function decline, and a higher risk for chronic liver and cardiovascular diseases in adults. Precise mechanisms of early developmental plasticity to intervene in poor fetal programming and adult disease remain largely elusive and warrant extensive research. Selecting natural piglets’ model of IUGR, using the liver as a readout and combining previous transcriptome findings, a map of cellular landscape was created to reveal a sex-dependent manner in IUGR-induced hepatic injury and its long-term functional repercussions.Here, we show data on the transcriptional profiles of 41,969 high-quality cells from normal birthweight (NBWs) and IUGR piglets (IUGRs) from hepatic tissue and demonstrated strong homology with human using human-derived liver single-cell dataset. We discovered that male liver was much more severely damaged and inflammation by IUGR than female liver at the one-week postnatal node.
Project description:Perinatal nutritional imbalances may have long-lasting consequences on health and disease, increasing risk of obesity, insulin resistance, type 2 diabetes or cardiovascular disease. This idea has been conceptualized in the Developmental Origins of Health and Disease Hypothesis (DOHaD). In addition, there is evidence that such early-programmed phenotypes can be transmitted to the following generation(s). It is proposed that, environmentally induced, transmission of disease risk is mediated by epigenetic mechanisms. The aim of this study was to determine whether patterns of gene expression in the first generation offspring are also present in the following generation offspring, via the paternal lineage. Paternal transmission of patterns of gene expression strongly suggest epigenetic inheritance of disease risk. Liver tissue was obtained from the follwing experimental groups: a) control male mice, b) adult male mice previously exposed to 50% caloric restriction in utero (IUGR), c) adult male mice overfed during lactation (ON), d) adult male offspring from control mice, e) adult male offspring from IUGR mice and f) adult male offspring from ON mice.RNA was extracted and processed for further hybridization on Affymetrix microarrays (GeneChip Mouse Genome 430 2.0 (Affymetrix, Santa Clara, CA)).
Project description:The aim of this research is to reveal the cellular crosstalk in fibrosis liver using transgenic pigs (TG) expressing humanized risk genes (PNPLA3I148M-GIPRdn-hIAPP) as a MAFLD model. The study uses single-nucleus sequencing to reveal the differentiation and interaction characteristics of various cell populations in the liver during the development of MAFLD. Compared to wild-type pigs (WT), the model pigs showed significantly increased plasma levels of neurotensin and phosphatidylserine, along with insulin resistance and significantly decreased HDL-c levels. After 6 months of high-fat high-sucrose diet induction, the model pigs exhibited obvious liver pathological features, including fat deposition, inflammatory cell aggregation, fibrosis, and blocked insulin signaling pathways. Single-nucleusl transcriptome sequencing identified six main cell types in the liver, and correlation analysis showed similar liver cell clusters, zones, and functions between pigs and humans. Liver cells near the central vein and liver sinusoidal endothelial cells (LSECs) were sensitive to metabolic changes and exhibited impaired function and reduced numbers first. Fibrosis-related pathways and the Rap1 pathway were activated in hepatic stellate cells of the model pigs, while retinol metabolism decreased, and the number of activated hepatic stellate cells increased. The differentiation direction of macrophages in the model pig's liver was markedly different from that in the WT pigs; the former differentiated toward the M1 phenotype, showing high expression of MHC-II antigen presentation molecules, various cytokines, phagosomes, and lysosomal-related genes. In contrast, the macrophages in the WT pig's liver primarily differentiated toward the M2 phenotype and expressed genes related to immune regulation and injury repair. There was active cell interaction between hepatic stellate cells, endothelial cells, and M1-type macrophages, promoting the development of chronic inflammation and fibrosis through interactions of receptors such as FGFs-FGFRs, PDGFs-PDGFRs, EFNA1-EPHRs, and CXCL12-CXCR4/CXCR7.In conclusion, the transgenic pig model of MAFLD exhibits liver pathological features consistent with MAFLD patients. The receptor interactions between hepatic stellate cells, endothelial cells, and macrophages play a key role in regulating metabolic inflammation and fibrosis development. Remarkable changes in receptor interactions hold promise for application in MAFLD drug development.
Project description:To evaluate functional consequences of insulin-deficient diabetes mellitus for the liver, we used a genetically engineered pig model of mutant INS gene induced diabetes of youth (MIDY). Liver samples of MIDY pigs and wild-type (WT) littermate controls were analyzed by label-free proteomics to reveal pathways and key drivers significantly affected by chronic insulin deficiency and hyperglycemia.
Project description:<p>Intrauterine growth restriction (IUGR) is associated with increased relative liver weight at birth, hepatic function decline and a higher risk for chronic liver and cardiovascular diseases in adults. Precise mechanisms of early developmental plasticity to intervene in poor fetal programming and adult disease remain largely elusive and warrant extensive research. Selecting natural piglets’ model of IUGR, using the liver as a readout and combining previous transcriptome findings, a map of cellular landscape was created to reveal a sex-dependent manner in IUGR-induced hepatic injury and its long-term functional repercussions. At the one-week postnatal node, we reveal that in contrast to a converting trend about an immune-adapted phenotype in female IUGRs, the more severe hepatic injury in male IUGRs is attributed to persistent inflammation caused by abnormal lipid metabolism, which disrupts the interactions of non-immune cells, resulting in impaired liver regeneration lasting into adulthood. In addition, we consider APOA4 as a new biomarker for the detection of IUGRs and discover its underlying role in hypoxic conditions. Our study opens the possibility to early therapy and screening of gender-specific hepatic injury brought on by IUGR.</p>
Project description:During the perinatal period, unique metabolic adaptations support energetic requirements for rapid growth. To gain insight into perinatal adaptations, quantitative proteomics were performed comparing the livers of yorkshire pigs at postnatal day seven and adult. These data revealed differences in the metabolic control of liver function including significant changes in lipid and carbohydrate metabolic pathways. Newborn livers showed an enrichment of proteins in lipid catabolism and gluconeogenesis concomitant with elevated liver carnitine and acylcarnitines levels. Sugar kinases were some of the most dramatically differentially enriched proteins comparing neonatal and adult pigs including galactokinase 1 (Galk1), ketohexokinase (KHK), hexokinase 1 (HK1) and hexokinase 4 (GCK). Interestingly, hexokinase domain containing 1 (HKDC1), an enigmatic fifth hexokinase associated with glucose disturbances in pregnant women was highly enriched in the liver during the prenatal and perinatal periods and continuously declined throughout postnatal development in pigs and mice. These changes were confirmed via Western blot and mRNA expression. These data provide new insights into the developmental and metabolic adaptations in the liver during the transition from the perinatal period to adulthood in multiple mammalian species.
Project description:The present study was designed to test the hypothesis that limited growth of the fetal liver in the model of maternal fasting is independent of well-characterized signaling mechanisms that are known to regulate somatic growth in adult animals. We profiled the fetal hepatic transcriptome and translatome in control (n=3) and IUGR (n=3) fetuses. To induce IUGR, pregnant dams were fasted for 48hr starting on E17, term being 21 days. Fetal rats were delivered by C-section and livers were removed. All livers were flash frozen in liquid nitrogen. RNA was extracted from polysomes that were generated using sucrose density fraction, as well as total liver. RNA was hybridized to Affymetrix GeneChip Rat Gene 1.0 ST Arrays.
Project description:Intrauterine growth restriction (IUGR) impairs fetal growth and development, perturbs nutrient metabolism, and increases the risk of developing diseases in the postnatal life. However, the underlying mechanisms by which IUGR affects fetuses remain incompletely understood. Here, we applied high-throughput proteomics approach and biochemical analysis to investigate the impact of IUGR on fetal liver.