Project description:Mutations in the E3 ubiquitin ligase Mkrn3 are associated with precocious puberty in humans. In order to determine the targets of Mkrn3, we performed a TMT-based proteomic analysis of Mkrn3 WT vs KO mouse brains.
Project description:MKRN3, whose deletion or loss-of-function mutations were genetically associated with human centeral precocious puberty (CPP). To identify the potential substrates for MKRN3, MKRN3 was transfected into HEK293T cells as bait, and its interacting proteins were identified by mass spectrum and functions were extensively studied.
Project description:Hypothalamic hamartomas (HHs) are congenital lesions of the neuroendocrine brain composed of neurons and astroglia. Frequently, HHs are associated with central precocious puberty (CPP) and/or gelastic seizures. Because HHs might express genes similar to those required for the initiation of normal puberty we used cDNA arrays to compare the gene expression profile of a HH associated with CPP with three HHs not accompanied by sexual precocity. Our aim was to identify genes whose expression may be selectively altered in the HH with CPP and hence, involved in the onset of puberty. Experiment Overall Design: Affymetrix arrays were used to detect global changes in gene expression. The results of this analysis were confirmed by semi-quantitative PCR.
Project description:Hypothalamic hamartomas (HHs) are congenital lesions of the neuroendocrine brain composed of neurons and astroglia. Frequently, HHs are associated with central precocious puberty (CPP) and/or gelastic seizures. Because HHs might express genes similar to those required for the initiation of normal puberty we used cDNA arrays to compare the gene expression profile of a HH associated with CPP with three HHs not accompanied by sexual precocity. Our aim was to identify genes whose expression may be selectively altered in the HH with CPP and hence, involved in the onset of puberty. Keywords: comparison between HH with and without CPP
Project description:Obesity-associated asthma is recognized as a distinct entity with non-atopic T-helper 1 polarized systemic inflammation. DNA methylation is linked with T helper cell maturation and is associated with inflammatory patterns in asthma and obesity. However, it is unknown whether pathologic dysregulation of DNA methylation patterns occurs in obesity-associated asthma. Using HELP-tagging, we studied epigenome wide DNA methylation in peripheral blood mononuclear cells in 8 urban minority obese asthmatic pre-adolescent children and compared it to methylation in groups of 8 children with asthma alone, obesity alone and healthy controls. Ingenuity Pathway Analysis was used to identify biological pathways that were differentially targeted by methylation dysregulation. We found that obese asthmatics had distinct epigenome wide methylation patterns associated with decreased promoter methylation of a subset of genes, including RANTES, IL-12R and TBX21 and increased promoter methylation of CD23, a low affinity receptor for IgE and of TGFM-NM-2, inhibitor of Th cell activation. T cell signaling and macrophage activation were the two primary pathways that were selectively hypomethylated in obese asthmatics. These methylation patterns suggest that methylation is associated with non-atopic inflammation observed in obese asthmatic children compared to children with asthma alone and obesity alone. Our findings suggest a role of DNA methylation in the observed inflammatory patterns in pediatric obesity-associated asthma in minorities. 32 HpaII test
Project description:Makorin ring finger protein 3 (MKRN3) was identified as an inhibitor of puberty initiation with the report of loss-of-function mutations in association with central precocious puberty. To investigate the roles and mechanisms of action of MKRN3 within the human hypothalamus, we used hypothalamic neurons derived from human induced pluripotent stem cells (hiPSCs). MKRN3 deletion was introduced into hiPSCs using CRISPR interference (CRISPRi) technology. Three separated hypothalamic differentiation of MKRN3-wildtype hiPSCs (MKRN3-WT I, II and III) and MKRN3-deficient hiPSCs (MKRN3-KO I, II and III) were performed using an established hypothalamic neuron differentiation protocol. To identify hypothalamic targets of MKRN3, we performed comparative transcriptome analysis by RNA sequencing (RNA-seq) of MKRN3-WT and MKRN3-KO hypothalamic neurons after 30 days of differentiation.
2023-06-14 | GSE208722 | GEO
Project description:Gut microbiota and hormone interaction mediates precocious puberty in HFD mice
