Bonner, Caroline Farrelly, Angela M. Concannon, Caoimhín G. Düssmann, Heiko Baquié, Mathurin Virard, Isabelle Wobser, Hella Kögel, Donat Wollheim, Claes B. Rupnik, Marjan Byrne, Maria M. König, Hans-Georg Prehn, Jochen HM Bone morphogenetic protein 3 controls insulin gene expression and is down-regulated in INS-1 cells inducibly expressing a hepatocyte nuclear factor 1A-maturity-onset diabetes of the young mutation. <p>Inactivating mutations in the transcription factor hepatocyte nuclear factor (HNF) 1A cause HNF1A-maturity-onset diabetes of the young (HNF1A-MODY), the most common monogenic form of diabetes. To examine HNF1A-MODY-induced defects in gene expression, we performed a microarray analysis of the transcriptome of rat INS-1 cells inducibly expressing the common hot spot HNF1A frameshift mutation, Pro291fsinsC-HNF1A. Real-time quantitative PCR (qPCR), Western blotting, immunohistochemistry, reporter assays, and chromatin immunoprecipitation (ChIP) were used to validate alterations in gene expression and to explore biological activities of target genes. Twenty-four hours after induction of the mutant HNF1A protein, we identified a prominent down-regulation of the bone morphogenetic protein 3 gene (Bmp-3) mRNA expression. Reporter assays, qPCR, and Western blot analysis validated these results. In contrast, inducible expression of wild-type HNF1A led to a time-dependent increase in Bmp-3 mRNA and protein levels. Moreover, reduced protein levels of BMP-3 and insulin were detected in islets of transgenic HNF1A-MODY mice. Interestingly, treatment of naïve INS-1 cells or murine organotypic islet cultures with recombinant human BMP-3 potently increased their insulin levels and restored the decrease in SMAD2 phosphorylation and insulin gene expression induced by the HNF1A frameshift mutation. Our study suggests a critical link between HNF1A-MODY-induced alterations in Bmp-3 expression and insulin gene levels in INS-1 cells and indicates that the reduced expression of growth factors involved in tissue differentiation may play an important role in the pathophysiology of HNF1A-MODY.</p> Animals;Bone Morphogenetic Protein 3;Cell Line;Tumor;Down-Regulation;Frameshift Mutation;Gene Expression Profiling;Hepatocyte Nuclear Factor 1-alpha;Humans;Insulin;Insulin-Secreting Cells;Mice;Oligonucleotide Array Sequence Analysis;Promoter Regions;Genetic;Rats;Physiology;Medical Physics 2019-11-22
    https://repository.rcsi.com/articles/journal_contribution/Bone_morphogenetic_protein_3_controls_insulin_gene_expression_and_is_down-regulated_in_INS-1_cells_inducibly_expressing_a_hepatocyte_nuclear_factor_1A-maturity-onset_diabetes_of_the_young_mutation_/10792919