BACKGROUND: Nonalcoholic fatty liver disease (NAFLD) is increasingly prevalent and represents a growing challenge in terms of prevention and treatment. A minority of affected patients develops inflammation, subsequently fibrosis, cirrhosis and hepatocellular carcinoma (HCC). HCC is a leading cause of cancer-related death. An increased number of senescent cells correlate with age-related tissue degeneration during NAFLD-induced HCC. Senolytics are promising agents that target selectively senescent cells. Previous studies showed that whereas a combination of the senolytic drugs dasatinib and quercetin (D + Q) reduced NAFLD in mice, D + Q lacked efficacy in removing doxorubicin-induced β-gal-positive senescent cells in human HCC xenografted mice. Whether D + Q has an effect on the age-associated spectrum of NAFLD-inflammation-HCC remains unknown. METHODS: Here, we utilized an established model of age- and obesity-associated HCC, the low dose diethylnitrosamine (DEN)/high fat diet (HFD), a regimen promoting liver inflammation and tumorigenesis over a long period of 9 months. Four groups of mice each were created: group 1 included control untreated mice; group 2 included mice treated with D + Q; group 3 included mice undergoing the DEN/HFD protocol; group 4 included mice undergoing the DEN/HFD protocol with the administration of D + Q. At the end of the chemical/dietary regimen, we analyzed liver damage and cell senescence by histopathology, qPCR and immunoblotting approaches. RESULTS: Unexpectedly, D + Q worsened liver disease progression in the DEN/HFD mouse model, slightly increasing histological damage and tumorigenesis, while having no effect on senescent cells removal. CONCLUSIONS: In summary, using an animal model that fully recapitulates NAFLD, we demonstrate that these compounds are ineffective against age-associated NAFLD-induced HCC. Video Abstract.
- Klíčová slova
- Cancer, Inflammation, Liver diseases, Obesity, Senolytics,
- MeSH
- dasatinib škodlivé účinky MeSH
- dieta s vysokým obsahem tuků MeSH
- diethylnitrosamin MeSH
- léky proti stárnutí škodlivé účinky MeSH
- modely nemocí na zvířatech MeSH
- myši inbrední C57BL MeSH
- myši MeSH
- nealkoholová steatóza jater krev genetika patologie MeSH
- nemoci jater krev genetika patologie MeSH
- obezita krev genetika patologie MeSH
- progrese nemoci * MeSH
- quercetin škodlivé účinky MeSH
- regulace genové exprese MeSH
- stárnutí genetika patologie MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- dasatinib MeSH
- diethylnitrosamin MeSH
- léky proti stárnutí MeSH
- quercetin MeSH
Lipid catabolism and anabolism changes play a role in stemness acquisition by cancer cells, and cancer stem cells (CSCs) are particularly dependent on the activity of the enzymes involved in these processes. Lipidomic changes could play a role in CSCs' ability to cause disease relapse and chemoresistance. The exploration of lipid composition and metabolism changes in CSCs in the context of hepatocellular cancer (HCC) is still incomplete and their lipidomic scenario continues to be elusive. We aimed to evaluate through high-throughput mass spectrometry (MS)-based lipidomics the levels of the members of the six major classes of sphingolipids and phospholipids in two HCC cell lines (HepG2 and Huh-7) silenced for the expression of histone variant macroH2A1 (favoring stemness acquisition), or silenced for the expression of focal adhesion tyrosine kinase (FAK) (hindering aggressiveness and stemness). Transcriptomic changes were evaluated by RNA sequencing as well. We found definite lipidomic and transcriptomic changes in the HCC lines upon knockdown (KD) of macroH2A1 or FAK, in line with the acquisition or loss of stemness features. In particular, macroH2A1 KD increased total sphingomyelin (SM) levels and decreased total lysophosphatidylcholine (LPC) levels, while FAK KD decreased total phosphatidylcholine (PC) levels. In conclusion, in HCC cell lines knocked down for specific signaling/epigenetic processes driving opposite stemness potential, we defined a lipidomic signature that hallmarks hepatic CSCs to be exploited for therapeutic strategies.
- Klíčová slova
- FAK, HCC, cancer stem cells, macroH2A1, stemness,
- MeSH
- buňky Hep G2 MeSH
- fokální adhezní kinasa 1 antagonisté a inhibitory nedostatek genetika MeSH
- fosfatidylcholiny metabolismus MeSH
- genový knockdown MeSH
- hepatocelulární karcinom genetika metabolismus patologie MeSH
- histony antagonisté a inhibitory nedostatek genetika MeSH
- lidé MeSH
- lipidomika MeSH
- lysofosfatidylcholiny metabolismus MeSH
- metabolismus lipidů * genetika MeSH
- nádorové biomarkery genetika metabolismus MeSH
- nádorové buněčné linie MeSH
- nádorové kmenové buňky metabolismus patologie MeSH
- nádory jater genetika metabolismus patologie MeSH
- regulace genové exprese u nádorů MeSH
- sekvenování transkriptomu MeSH
- sfingomyeliny metabolismus MeSH
- stanovení celkové genové exprese MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- fokální adhezní kinasa 1 MeSH
- fosfatidylcholiny MeSH
- histony MeSH
- lysofosfatidylcholiny MeSH
- MACROH2A1 protein, human MeSH Prohlížeč
- nádorové biomarkery MeSH
- PTK2 protein, human MeSH Prohlížeč
- sfingomyeliny MeSH
BACKGROUND & AIMS: Growth Differentiation Factor 11 (GDF11) is an anti-aging factor, yet its role in liver diseases is not established. We evaluated the role of GDF11 in healthy conditions and in the transition from non-alcoholic fatty liver disease (NAFLD) to non-alcoholic steatohepatitis (NASH). RESULTS: GDF11 mRNA levels positively correlated with NAFLD activity score and with CPT1, SREBP, PPARγ and Col1A1 mRNA levels, and associated to portal fibrosis, in morbidly obese patients with NAFLD/NASH. GDF11-treated mice showed mildly exacerbated hepatic collagen deposition, accompanied by weight loss and without changes in liver steatosis or inflammation. GDF11 triggered ALK5-dependent SMAD2/3 nuclear translocation and the pro-fibrogenic activation of HSC. CONCLUSIONS: GDF11 supplementation promotes mild liver fibrosis. Even considering its beneficial metabolic effects, caution should be taken when considering therapeutics that regulate GDF11. METHODS: We analyzed liver biopsies from a cohort of 33 morbidly obese adults with NAFLD/NASH. We determined the correlations in mRNA expression levels between GDF11 and genes involved in NAFLD-to-NASH progression and with pathological features. We also exposed wild type or obese mice with NAFLD to recombinant GDF11 by daily intra-peritoneal injection and monitor the hepatic pathological changes. Finally, we analyzed GDF11-activated signaling pathways in hepatic stellate cells (HSC).
- Klíčová slova
- NAFLD, NASH, fibrosis, growth differentiation factor 11, liver,
- MeSH
- buněčné linie MeSH
- dospělí MeSH
- experimentální cirhóza jater chemicky indukované metabolismus patologie MeSH
- jaterní cirhóza diagnóza etiologie genetika metabolismus MeSH
- jaterní hvězdicovité buňky metabolismus patologie MeSH
- játra metabolismus patologie MeSH
- kostní morfogenetické proteiny genetika metabolismus toxicita MeSH
- lidé středního věku MeSH
- lidé MeSH
- morbidní obezita komplikace diagnóza MeSH
- myši inbrední C57BL MeSH
- nealkoholová steatóza jater diagnóza etiologie genetika metabolismus MeSH
- progrese nemoci MeSH
- růstové diferenciační faktory genetika metabolismus toxicita MeSH
- signální transdukce MeSH
- studie případů a kontrol MeSH
- zvířata MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- GDF11 protein, human MeSH Prohlížeč
- kostní morfogenetické proteiny MeSH
- růstové diferenciační faktory MeSH
OBJECTIVE: While circulating nucleosome levels are high in obese mouse models, it is unknown where these nucleosomes originate from and whether they are a marker of cardio-metabolic health in humans. Here, we aimed to determine whether an association exists between circulating nucleosomes and the risk of developing obesity, metabolic syndrome (MetS) and/or a dysfunctional cardiovascular performance. METHODS: We randomly selected 120 participants of the Kardiovize Brno 2030 study across three BMI strata: BMI 18-25, 25-30, and > 30. We assessed the association between circulating nucleosome levels and the risk of obesity, MetS, and poor cardiovascular health. We then cultured human neutrophils, adipocytes, and hepatoma cells to study nucleosome origins in a fat-rich environment. RESULTS: Circulating nucleosome levels positively correlated with BMI (R = 0.602, p < 0.05), fatty liver index (R = 0.622, p < 0.05), left ventricular mass (R = 0.457, p < 0.05), and associated with MetS (p < 0.001) and poor cardiovascular health (p < 0.001). Incubating neutrophils with 1-10 μM free fatty acids triggered nucleosome production without concomitant cell death. Nucleosomes were not produced during pre-adipocyte differentiation or upon incubation of hepatic cells with palmitic acid. CONCLUSIONS: Neutrophils are a bona fide source of circulating nucleosomes in an obesogenic environment and in overweight/obese patients. High nucleosome levels are associated with MetS and cardiovascular performance, and might represent novel candidate biomarkers for cardio-metabolic health.
- Klíčová slova
- Cardiovascular disease, Epigenetics, Liquid biopsy, Metabolic health, Nucleosome,
- MeSH
- buňky Hep G2 MeSH
- dospělí MeSH
- index tělesné hmotnosti MeSH
- kardiovaskulární nemoci etiologie MeSH
- kultivované buňky MeSH
- lidé středního věku MeSH
- lidé MeSH
- metabolický syndrom krev komplikace MeSH
- nadváha krev komplikace metabolismus MeSH
- neutrofily cytologie metabolismus MeSH
- nukleozomy metabolismus MeSH
- obezita krev komplikace metabolismus MeSH
- senioři MeSH
- tukové buňky cytologie metabolismus MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- nukleozomy MeSH
Maternal obesity predisposes offspring to metabolic dysfunction and Non-Alcoholic Fatty Liver Disease (NAFLD). Melanocortin-4 receptor (Mc4r)-deficient mouse models exhibit obesity during adulthood. Here, we aim to determine the influence of the Mc4r gene on the liver of mice subjected to perinatal diet-induced obesity. Female mice heterozygous for Mc4r fed an obesogenic or a control diet for 5 weeks were mated with heterozygous males, with the same diet continued throughout pregnancy and lactation, generating four offspring groups: control wild type (C_wt), control knockout (C_KO), obese wild type (Ob_wt), and obese knockout (Ob_KO). At 21 days, offspring were genotyped, weaned onto a control diet, and sacrificed at 6 months old. Offspring phenotypic characteristics, plasma biochemical profile, liver histology, and hepatic gene expression were analyzed. Mc4r_ko offspring showed higher body, liver and adipose tissue weights respect to the wild type animals. Histological examination showed mild hepatic steatosis in offspring group C_KO. The expression of hepatic genes involved in regulating inflammation, fibrosis, and immune cell infiltration were upregulated by the absence of the Mc4r gene. These results demonstrate that maternal obesogenic feeding during the perinatal period programs offspring obesity development with involvement of the Mc4r system.
- Klíčová slova
- Non-Alcoholic Fatty Liver Disease, developmental programming, intra-abdominal fat, maternal nutrition, obesity,
- MeSH
- alanintransaminasa krev MeSH
- aspartátaminotransferasy krev MeSH
- fyziologie výživy v mateřství * MeSH
- játra metabolismus MeSH
- krevní glukóza metabolismus MeSH
- modely nemocí na zvířatech MeSH
- myši knockoutované MeSH
- myši obézní MeSH
- myši MeSH
- obezita genetika MeSH
- perinatální péče MeSH
- receptor melanokortinový typ 4 nedostatek genetika MeSH
- regulace genové exprese MeSH
- těhotenství MeSH
- triglyceridy krev MeSH
- zpožděný efekt prenatální expozice genetika MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- těhotenství MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- alanintransaminasa MeSH
- aspartátaminotransferasy MeSH
- krevní glukóza MeSH
- MC4R protein, mouse MeSH Prohlížeč
- receptor melanokortinový typ 4 MeSH
- triglyceridy MeSH
Hepatic stellate cells (HSCs), also known as perisinusoidal cells, are pericytes found in the perisinusoidal space of the liver. HSCs are the major cell type involved in liver fibrosis, which is the formation of scar tissue in response to liver damage. When the liver is damaged, stellate cells can shift into an activated state, characterized by proliferation, contractility and chemotaxis. The activated HSCs secrete collagen scar tissue, which can lead to cirrhosis. Recent studies have shown that in vivo activation of HSCs by fibrogenic agents can eventually lead to senescence of these cells, which would contribute to reversal of fibrosis although it may also favor the insurgence of liver cancer. HSCs in their non-active form store huge amounts of retinoic acid derivatives in lipid droplets, which are progressively depleted upon cell activation in injured liver. Retinoic acid is a metabolite of vitamin A (retinol) that mediates the functions of vitamin A, generally required for growth and development. The precise function of retinoic acid and its alterations in HSCs has yet to be elucidated, and nonetheless in various cell types retinoic acid and its receptors (RAR and RXR) are known to act synergistically with peroxisome proliferator-activated receptor gamma (PPAR-gamma) signaling through the activity of transcriptional heterodimers. Here, we review the recent advancements in the understanding of how retinoic acid signaling modulates the fibrogenic potential of HSCs and proposes a synergistic combined action with PPAR-gamma in the reversal of liver fibrosis.
- Klíčová slova
- Fibrogenesis, Hepatic stellate cells, Retinoic acid,
- MeSH
- jaterní cirhóza patofyziologie MeSH
- jaterní hvězdicovité buňky fyziologie MeSH
- keratolytika MeSH
- lidé MeSH
- PPAR gama metabolismus MeSH
- protinádorové látky MeSH
- signální transdukce * MeSH
- tretinoin metabolismus MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
- Názvy látek
- keratolytika MeSH
- PPAR gama MeSH
- protinádorové látky MeSH
- tretinoin MeSH
We investigated the regulation of hepatic ER stress in healthy liver and adult or perinatally programmed diet-induced non-alcoholic fatty liver disease (NAFLD). Female mice were fed either obesogenic or control diet before mating, during pregnancy and lactation. Post-weaning, offspring from each maternal group were divided into either obesogenic or control diet. At six months, offspring were sacrificed at 4-h intervals over 24 h. Offspring fed obesogenic diets developed NAFLD phenotype, and the combination of maternal and offspring obesogenic diets exacerbated this phenotype. UPR signalling pathways (IREα, PERK, ATF6) and their downstream regulators showed different basal rhythmicity, which was modified in offspring exposed to obesogenic diet and maternal programming. The double obesogenic hit increased liver apoptosis measured by TUNEL staining, active caspase-3 and phospho-JNK and GRP78 promoter methylation levels. This study demonstrates that hepatic UPR is rhythmically activated. The combination of maternal obesity (MO) and obesogenic diets in offspring triggered altered UPR rhythmicity, DNA methylation and cellular apoptosis.
- Klíčová slova
- ER stress, NAFLD, Obesity, circadian rhythm, developmental programming,
- MeSH
- chaperon endoplazmatického retikula BiP MeSH
- dieta s vysokým obsahem tuků škodlivé účinky MeSH
- endoplazmatické retikulum fyziologie MeSH
- fyziologický stres účinky léků MeSH
- homeostáza MeSH
- játra účinky léků MeSH
- krmivo pro zvířata analýza MeSH
- myši inbrední C57BL MeSH
- myši MeSH
- obezita chemicky indukované metabolismus MeSH
- těhotenství MeSH
- zpožděný efekt prenatální expozice MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- těhotenství MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
The prevalence of non-alcoholic fatty pancreas disease (NAFPD) is increasing in parallel with obesity rates. Stress-related alterations in endoplasmic reticulum (ER), such as the unfolded protein response (UPR), are associated with obesity. The aim of this study was to investigate ER imbalance in the pancreas of a mice model of adult and perinatal diet-induced obesity. Twenty female C57BL/6J mice were assigned to control (Con) or obesogenic (Ob) diets prior to and during pregnancy and lactation. Their offspring were weaned onto Con or Ob diets up to 6 months post-partum. Then, after sacrifice, plasma biochemical analyses, gene expression, and protein concentrations were measured in pancreata. Offspring of Ob-fed mice had significantly increased body weight (p < 0.001) and plasma leptin (p < 0.001) and decreased insulin (p < 0.01) levels. Maternal obesogenic diet decreased the total and phosphorylated Eif2α and increased spliced X-box binding protein 1 (XBP1). Pancreatic gene expression of downstream regulators of UPR (EDEM, homocysteine-responsive endoplasmic reticulum-resident (HERP), activating transcription factor 4 (ATF4), and C/EBP homologous protein (CHOP)) and autophagy-related proteins (LC3BI/LC3BII) were differently disrupted by obesogenic feeding in both mothers and offspring (from p < 0.1 to p < 0.001). Maternal obesity and Ob feeding in their offspring alter UPR in NAFPD, with involvement of proapoptotic and autophagy-related markers. Upstream and downstream regulators of PERK, IRE1α, and ATF6 pathways were affected differently following the obesogenic insults.
- Klíčová slova
- Autophagy, ER stress, Fatty pancreas, Obesity, Perinatal programming, UPR,
- MeSH
- autofagie MeSH
- biologické markery krev metabolismus MeSH
- dieta s vysokým obsahem tuků škodlivé účinky MeSH
- fyziologie výživy v mateřství * MeSH
- inzulin krev MeSH
- komplikace těhotenství etiologie patofyziologie MeSH
- konzumní sacharóza škodlivé účinky MeSH
- laktace MeSH
- leptin krev MeSH
- myši inbrední C57BL MeSH
- obezita etiologie patofyziologie MeSH
- odstavení MeSH
- pankreas imunologie metabolismus patofyziologie MeSH
- pankreatitida etiologie imunologie metabolismus patofyziologie MeSH
- signální dráha UPR * MeSH
- stres endoplazmatického retikula * MeSH
- těhotenství MeSH
- vývojová regulace genové exprese MeSH
- zvířata MeSH
- Check Tag
- těhotenství MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- srovnávací studie MeSH
- Názvy látek
- biologické markery MeSH
- inzulin MeSH
- konzumní sacharóza MeSH
- leptin MeSH
