转录因子HNF1A、HNF4A和FOXA2调节肝细胞蛋白质N-糖基化  

Transcription Factors HNF1A,HNF4A,and FOXA2 Regulate Hepatic Cell Protein N-Glycosylation

作　　者：Vedrana Vicic Bockor Nika Foglar Goran Josipovic Marija Klasic Ana Vujic Branimir Plavsa Toma Keser Samira Smajlovic Aleksandar Vojta Vlatka Zoldos 

机构地区：[1]Laboratory for Epigenetics,Department of Biology,Faculty of Science,University of Zagreb,Zagreb 10000,Croatia [2]Genos Glycoscience Research Laboratory,Zagreb 10000,Croatia [3]Faculty of Pharmacy and Biochemistry,University of Zagreb,Zagreb 10000,Croatia

出　　处：《Engineering》2024年第1期57-68,共12页工程（英文）

基　　金：the European Structural and Investment Funded Grant"Cardio Metabolic"(#KK.01.2.1.02.0321);the Croatian National Centre of Research Excellence in Personalized Healthcare Grant(#KK.01.1.1.01.0010);the European Regional Development Fund Grant,project"CRISPR/Cas9-CasMouse"(#KK.01.1.1.04.0085);the European Structural and Investment Funded Project of Centre of Competence in Molecular Diagnostics(#KK.01.2.2.03.0006);the Croatian National Centre of Research Excellence in Personalized Healthcare Grant(#KK.01.1.1.01.0010).

摘　　要：Hepatocyte nuclear factor 1 alpha(HNF1A),hepatocyte nuclear factor 4 alpha(HNF4A),and forkhead box protein A2(FOXA2)are key transcription factors that regulate a complex gene network in the liver,cre-ating a regulatory transcriptional loop.The Encode and ChIP-Atlas databases identify the recognition sites of these transcription factors in many glycosyltransferase genes.Our in silico analysis of HNF1A,HNF4A.and FOXA2 binding to the ten candidate glyco-genes studied in this work confirms a significant enrich-ment of these transcription factors specifically in the liver.Our previous studies identified HNF1A as a master regulator of fucosylation,glycan branching,and galactosylation of plasma glycoproteins.Here,we aimed to functionally validate the role of the three transcription factors on downstream glyco-gene transcriptional expression and the possible effect on glycan phenotype.We used the state-of-the-art clus-tered regularly interspaced short palindromic repeats/dead Cas9(CRISPR/dCas9)molecular tool for the downregulation of the HNF1A,HNF4A,and FOXA2 genes in HepG2 cells-a human liver cancer cell line.The results show that the downregulation of all three genes individually and in pairs affects the transcrip-tional activity of many glyco-genes,although downregulation of glyco-genes was not always followed by an unambiguous change in the corresponding glycan structures.The effect is better seen as an overall change in the total HepG2 N-glycome,primarily due to the extension of biantennary glycans.We propose an alternative way to evaluate the N-glycome composition via estimating the overall complexity of the glycome by quantifying the number of monomers in each glycan structure.We also propose a model showing feedback loops with the mutual activation of HNF1A-FOXA2 and HNF4A-FOXA2 affecting glyco-genes and protein glycosylation in HepG2 cells.

关 键 词：Clustered regularly interspaced short palindromic repeats/dead Cas9(CRISPR/dCas9) EPIGENETICS Hepatocyte nuclear factor 1 alpha(HNF1A) Hepatocyte nuclear factor 4 alpha(HNF4A) Forkhead box protein A2(FOXA2) N-GLYCOSYLATION HepG2 cells 

分 类 号：R73[医药卫生—肿瘤]