Similar Datasets
Project description:Growth hormone (GH) signaling through STAT5B is a central regulator of hepatic metabolism, yet the functional consequences of disease-associated STAT5B variants remain poorly understood. Here, we generated mice carrying STAT5BY665F (gain-of-function) and STAT5BY665H (loss-of-function) alleles using CRISPR/Cas9-mediated base editing to dissect their impact on metabolic regulation. STAT5BY665F mutants developed hepatic lipid accumulation, hypercholesterolemia, and enhanced insulin sensitivity, whereas STAT5BY665H mice displayed reduced body weight and impaired insulin responsiveness. Transcriptomic analyses revealed that STAT5BY665F strongly activated lipid, cholesterol, and immune transcriptional programs, while STAT5BY665H failed to induce these pathways. ChIP-seq demonstrated extensive STAT5BY665F enhancer occupancy at metabolic and immune loci, contrasting with the minimal chromatin engagement of STAT5BY665H. Beyond the liver, STAT5BY665F broadly reprogrammed adipose tissue gene expression, activating lipid metabolism and immune regulatory networks, whereas STAT5BY665H exerted more restricted effects. Notably, transcriptional and chromatin remodeling by STAT5B variants displayed sex-dependent differences, with male livers showing more pronounced reprogramming than females. Together, these findings illustrate how alterations in STAT5B activity that alter enhancer activation led to altered metabolic function in liver and adipose tissue.
2026-02-12 | GSE310017 | GEO
Project description:Growth hormone (GH) signaling through STAT5B is a central regulator of hepatic metabolism, yet the functional consequences of disease-associated STAT5B variants remain poorly understood. Here, we generated mice carrying STAT5BY665F (gain-of-function) and STAT5BY665H (loss-of-function) alleles using CRISPR/Cas9-mediated base editing to dissect their impact on metabolic regulation. STAT5BY665F mutants developed hepatic lipid accumulation, hypercholesterolemia, and enhanced insulin sensitivity, whereas STAT5BY665H mice displayed reduced body weight and impaired insulin responsiveness. Transcriptomic analyses revealed that STAT5BY665F strongly activated lipid, cholesterol, and immune transcriptional programs, while STAT5BY665H failed to induce these pathways. ChIP-seq demonstrated extensive STAT5BY665F enhancer occupancy at metabolic and immune loci, contrasting with the minimal chromatin engagement of STAT5BY665H. Beyond the liver, STAT5BY665F broadly reprogrammed adipose tissue gene expression, activating lipid metabolism and immune regulatory networks, whereas STAT5BY665H exerted more restricted effects. Notably, transcriptional and chromatin remodeling by STAT5B variants displayed sex-dependent differences, with male livers showing more pronounced reprogramming than females. Together, these findings illustrate how alterations in STAT5B activity that alter enhancer activation led to altered metabolic function in liver and adipose tissue.
2026-02-12 | GSE309955 | GEO
Project description:Growth hormone (GH) signaling through STAT5B is a central regulator of hepatic metabolism, yet the functional consequences of disease-associated STAT5B variants remain poorly understood. Here, we generated mice carrying STAT5BY665F (gain-of-function) and STAT5BY665H (loss-of-function) alleles using CRISPR/Cas9-mediated base editing to dissect their impact on metabolic regulation. STAT5BY665F mutants developed hepatic lipid accumulation, hypercholesterolemia, and enhanced insulin sensitivity, whereas STAT5BY665H mice displayed reduced body weight and impaired insulin responsiveness. Transcriptomic analyses revealed that STAT5BY665F strongly activated lipid, cholesterol, and immune transcriptional programs, while STAT5BY665H failed to induce these pathways. ChIP-seq demonstrated extensive STAT5BY665F enhancer occupancy at metabolic and immune loci, contrasting with the minimal chromatin engagement of STAT5BY665H. Beyond the liver, STAT5BY665F broadly reprogrammed adipose tissue gene expression, activating lipid metabolism and immune regulatory networks, whereas STAT5BY665H exerted more restricted effects. Notably, transcriptional and chromatin remodeling by STAT5B variants displayed sex-dependent differences, with male livers showing more pronounced reprogramming than females. Together, these findings illustrate how alterations in STAT5B activity that alter enhancer activation led to altered metabolic function in liver and adipose tissue.
2026-02-12 | GSE309954 | GEO
Project description:Disease-associated mutations on the STAT5B SH2 domain regulate cholesterol and lipid metabolism in liver
| PRJNA1364991 | ENA
Project description:Disease-associated mutations on the STAT5B SH2 domain regulate cholesterol and lipid metabolism in liver [RNA-Seq liver]
| PRJNA1365013 | ENA
Project description:Disease-associated mutations on the STAT5B SH2 domain regulate cholesterol and lipid metabolism in liver [ChIP-seq]
| PRJNA1364038 | ENA
Project description:Disease-associated mutations on the STAT5B SH2 domain regulate cholesterol and lipid metabolism in liver [RNA-seq]
| PRJNA1364040 | ENA
Project description:OBJECTIVE:Increases in hepatic and plasma cholesterol occur in patients with nonalcoholic fatty liver disease (NAFLD), although the reason for this is not well understood. We investigated whether Protease-Activated Receptor 2 (PAR2) plays a role in cholesterol and lipid homeostasis in NAFLD. METHODS:Human liver biopsies (n = 108) were quantified for PAR2 expression from NAFLD cases randomly selected and stratified by liver fibrosis stage, the primary predictor for clinical outcomes, while controlling for age, gender, and BMI between fibrosis groups. Demographic data and laboratory studies on plasma samples were obtained within 6 months of liver biopsy. Wild-type and PAR2-KO (C57BL/6 F2rl1-/-) mice were fed either normal or high fat diet for 16 weeks and plasma and liver assayed for lipids and soluble metabolites. RESULTS:Severity of NAFLD and plasma cholesterol levels significantly correlated with hepatocyte PAR2 expression in NAFLD patients. Conversely, PAR2 deficiency in mice resulted in reduced expression of key hepatic genes involved in cholesterol synthesis, a 50% drop in plasma and total liver cholesterol, and induced a reverse cholesterol transport system that culminated in 25% higher fecal bile acid output. PAR2-deficient mice exhibited enhanced fatty acid β-oxidation with a ketogenic shift and an unexpected increase in liver glycogenesis. Mechanistic studies identified Gi-Jnk1/2 as key downstream effectors of protease-activated PAR2 in the regulation of lipid and cholesterol homeostasis in liver. CONCLUSIONS:These data indicate that PAR2 may be a new target for the suppression of plasma cholesterol and hepatic fat accumulation in NAFLD and related metabolic conditions.
| S-EPMC6742970 | biostudies-literature
Project description:The point mutation N642H of the signal transducer and activator of transcription 5B (STAT5B) protein is associated with aggressive and drug-resistant forms of leukemia. This mutation is thought to promote cancer due to hyperactivation of STAT5B caused by increased stability of the active, parallel dimer state. However, the molecular mechanism leading to this stabilization is not well understood as there is currently no structure of the parallel dimer. To investigate the mutation's mechanism of action, we conducted extensive all-atom molecular dynamics simulations of multiple oligomeric forms of both STAT5B and STAT5BN642H, including a model for the parallel dimer. The N642H mutation directly affects the hydrogen bonding network within the phosphotyrosine (pY)-binding pocket of the parallel dimer, enhancing the pY-binding interaction. The simulations indicate that apo STAT5B is highly flexible, exploring a diverse conformational space. In contrast, apo STAT5BN642H accesses two distinct conformational states, one of which resembles the conformation of the parallel dimer. The simulation predictions of the effects of the mutation on structure and dynamics are supported by the results of hydrogen-deuterium exchange (HDX) mass spectrometry measurements carried out on STAT5B and STAT5BN642H in which a phosphopeptide was used to mimic the effects of parallel dimerization on the SH2 domain. The molecular-level information uncovered in this work contributes to our understanding of STAT5B hyperactivation by the N642H mutation and could help pave the way for novel therapeutic strategies targeting this mutation.
| S-EPMC11670306 | biostudies-literature
Project description:STAT5B is a vital transcription factor for lymphocytes. Here, function of two STAT5B mutations from human T cell leukemias: one substituting tyrosine 665 with phenylalanine (STAT5BY665F), the other with histidine (STAT5BY665H) was interrogated. In silico modeling predicted divergent energetic effects on homodimerization with a range of pathogenicity. In primary T cells in vitro STAT5BY665F showed gain-of-function while STAT5BY665H demonstrated loss-of-function. Introducing the mutation into the mouse genome illustrated that the gain-of-function Stat5b Y665F mutation resulted in accumulation of CD8+ effector and memory and CD4+ regulatory T-cells, altering CD8+/CD4+ ratios. In contrast, STAT5BY665H 'knock-in' mice showed diminished CD8+ effector and memory and CD4+ regulatory T cells. In contrast to wild-type STAT5, the STAT5BY665F variant displayed greater STAT5 phosphorylation, DNA binding and transcriptional activity following cytokine activation while the STAT5BY665H variant resembled a null. The work exemplifies how joining in silico and in vivo studies of single nucleotides deepens our understanding of disease-associated variants, revealing structural determinants of altered function, defining mechanistic roles, and, specifically here, identifying a gain-of function variant that does not directly induce hematopoietic malignancy.
| S-EPMC11722272 | biostudies-literature