title: Alternative signaling network activation through different insulin receptor family members caused by pro-mitogenic antidiabetic insulin analogues in human mammary epithelial cells.
abstract: BACKGROUND
  Insulin analogues are designed to have improved pharmacokinetic parameters compared to regular human insulin. This provides a sustained control of blood glucose levels in diabetic patients. All novel insulin analogues are tested for their mitogenic side effects, however these assays do not take into account the molecular mode of action of different insulin analogues. Insulin analogues can bind the insulin receptor and the insulin-like growth factor 1 receptor with different affinities and consequently will activate different downstream signaling pathways.
  
  
  METHODS
  Here we used a panel of MCF7 human breast cancer cell lines that selectively express either one of the isoforms of the INSR or the IGF1R. We applied a transcriptomics approach to assess the differential transcriptional programs activated in these cells by either insulin, IGF1 or X10 treatment.
  
  
  RESULTS
  Based on the differentially expressed genes between insulin versus IGF1 and X10 treatment, we retrieved a mitogenic classifier gene set. Validation by RT-qPCR confirmed the robustness of this gene set. The translational potential of these mitogenic classifier genes was examined in primary human mammary cells and in mammary gland tissue of mice in an in vivo model. The predictive power of the classifier genes was evaluated by testing all commercial insulin analogues in the in vitro model and defined X10 and glargine as the most potent mitogenic insulin analogues.
  
  
  CONCLUSIONS
  We propose that these mitogenic classifier genes can be used to test the mitogenic potential of novel insulin analogues as well as other alternative molecules with an anticipated affinity for the IGF1R.
authors.affiliation: Division of Toxicology, Leiden Academic Centre for Drug Research, Leiden University, Einsteinweg 55, Leiden, 2333 CC, The Netherlands. basterbraak@gmail.com.
authors.forename: Bas
authors.initials: B
authors.lastname: ter Braak
authors.affiliation: Division of Toxicology, Leiden Academic Centre for Drug Research, Leiden University, Einsteinweg 55, Leiden, 2333 CC, The Netherlands. s.wink.3@lacdr.leidenuniv.nl.
authors.forename: Steven
authors.initials: S
authors.lastname: Wink
authors.affiliation: Division of Toxicology, Leiden Academic Centre for Drug Research, Leiden University, Einsteinweg 55, Leiden, 2333 CC, The Netherlands. e.koedoot@lacdr.leidenuniv.nl.
authors.forename: Esmee
authors.initials: E
authors.lastname: Koedoot
authors.affiliation: Division of Toxicology, Leiden Academic Centre for Drug Research, Leiden University, Einsteinweg 55, Leiden, 2333 CC, The Netherlands. c.pont@lacdr.leidenuniv.nl.
authors.forename: Chantal
authors.initials: C
authors.lastname: Pont
authors.affiliation: Medicines Evaluation Board (MEB), Graadt van Roggenweg 500, Utrecht, 3531 AH, The Netherlands. cl.siezen@cbg-meb.nl.
authors.forename: Christine
authors.initials: C
authors.lastname: Siezen
authors.affiliation: Division of Toxicology, Leiden Academic Centre for Drug Research, Leiden University, Einsteinweg 55, Leiden, 2333 CC, The Netherlands. jw.vd.laan@cbg-meb.nl.|Medicines Evaluation Board (MEB), Graadt van Roggenweg 500, Utrecht, 3531 AH, The Netherlands. jw.vd.laan@cbg-meb.nl.|Centre for Health Protection, National Institute for Public Health and the Environment (RIVM), Antonie van Leeuwenhoeklaan 9, Bilthoven, 3721 MA, The Netherlands. jw.vd.laan@cbg-meb.nl.
authors.forename: Jan Willem
authors.initials: JW
authors.lastname: van der Laan
authors.affiliation: Division of Toxicology, Leiden Academic Centre for Drug Research, Leiden University, Einsteinweg 55, Leiden, 2333 CC, The Netherlands. b.water@lacdr.leidenuniv.nl.
authors.forename: Bob
authors.initials: B
authors.lastname: van de Water
mesh_terms: D000818:Animals; D015703:Antigens, CD / Q000378:metabolism; D001943:Breast Neoplasms / Q000378:metabolism; D045744:Cell Line, Tumor; D003920:Diabetes Mellitus / Q000188:drug therapy / Q000378:metabolism; D004847:Epithelial Cells / Q000378:metabolism*; D005260:Female; D006801:Humans; D007004:Hypoglycemic Agents / Q000494:pharmacology*; D007328:Insulin / Q000378:metabolism*; D007334:Insulin-Like Growth Factor I / Q000378:metabolism; D061986:MCF-7 Cells; D042361:Mammary Glands, Human / Q000187:drug effects / Q000378:metabolism*; D051379:Mice; D017526:Receptor, IGF Type 1 / Q000378:metabolism*; D011972:Receptor, Insulin / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology*
chemical_list: D015703:Antigens, CD; D007004:Hypoglycemic Agents; D007328:Insulin; D007334:Insulin-Like Growth Factor I; C435941:INSR protein, human; D017526:Receptor, IGF Type 1; D011972:Receptor, Insulin