title: Rapamycin reduces fibroblast proliferation without causing quiescence and induces STAT5A/B-mediated cytokine production.
abstract: Rapamycin is a well-known inhibitor of the Target of Rapamycin (TOR) signaling cascade; however, the impact of this drug on global genome function and organization in normal primary cells is poorly understood. To explore this impact, we treated primary human foreskin fibroblasts with rapamycin and observed a decrease in cell proliferation without causing cell death. Upon rapamycin treatment chromosomes 18 and 10 were repositioned to a location similar to that of fibroblasts induced into quiescence by serum reduction. Although similar changes in positioning occurred, comparative transcriptome analyses demonstrated significant divergence in gene expression patterns between rapamycin-treated and quiescence-induced fibroblasts. Rapamycin treatment induced the upregulation of cytokine genes, including those from the Interleukin (IL)-6 signaling network, such as IL-8 and the Leukemia Inhibitory Factor (LIF), while quiescent fibroblasts demonstrated up-regulation of genes involved in the complement and coagulation cascade. In addition, genes significantly up-regulated by rapamycin treatment demonstrated increased promoter occupancy of the transcription factor Signal Transducer and Activator of Transcription 5A/B (STAT5A/B). In summary, we demonstrated that the treatment of fibroblasts with rapamycin decreased proliferation, caused chromosome territory repositioning and induced STAT5A/B-mediated changes in gene expression enriched for cytokines.
authors.affiliation: a Department of Food and Bioproduct Sciences ; University of Saskatchewan ; Saskatoon , Canada.|b Institute of Environment, Health and Societies; Brunel University; London , Uxbridge , United Kingdom.
authors.forename: Zoe E
authors.initials: ZE
authors.lastname: Gillespie
authors.affiliation: c Department of Computer Science ; University of Saskatchewan ; Saskatoon , Canada.
authors.forename: Kimberly
authors.initials: K
authors.lastname: MacKay
authors.affiliation: a Department of Food and Bioproduct Sciences ; University of Saskatchewan ; Saskatoon , Canada.
authors.forename: Michelle
authors.initials: M
authors.lastname: Sander
authors.affiliation: c Department of Computer Science ; University of Saskatchewan ; Saskatoon , Canada.
authors.forename: Brett
authors.initials: B
authors.lastname: Trost
authors.affiliation: d Department of Medicine ; Division of Respirology, Critical Care and Sleep Medicine; Royal University Hospital ; Saskatoon , Canada.
authors.forename: Wojciech
authors.initials: W
authors.lastname: Dawicki
authors.affiliation: a Department of Food and Bioproduct Sciences ; University of Saskatchewan ; Saskatoon , Canada.
authors.forename: Aruna
authors.initials: A
authors.lastname: Wickramarathna
authors.affiliation: d Department of Medicine ; Division of Respirology, Critical Care and Sleep Medicine; Royal University Hospital ; Saskatoon , Canada.
authors.forename: John
authors.initials: J
authors.lastname: Gordon
authors.affiliation: c Department of Computer Science ; University of Saskatchewan ; Saskatoon , Canada.
authors.forename: Mark
authors.initials: M
authors.lastname: Eramian
authors.affiliation: b Institute of Environment, Health and Societies; Brunel University; London , Uxbridge , United Kingdom.
authors.forename: Ian R
authors.initials: IR
authors.lastname: Kill
authors.affiliation: b Institute of Environment, Health and Societies; Brunel University; London , Uxbridge , United Kingdom.
authors.forename: Joanna M
authors.initials: JM
authors.lastname: Bridger
authors.affiliation: c Department of Computer Science ; University of Saskatchewan ; Saskatoon , Canada.
authors.forename: Anthony
authors.initials: A
authors.lastname: Kusalik
authors.affiliation: e Department of Cell and Systems Biology ; University of Toronto ; Toronto , Canada.|f Centre for the Analysis of Genome Evolution and Function; University of Toronto , Toronto , ON , Canada.
authors.forename: Jennifer A
authors.initials: JA
authors.lastname: Mitchell
authors.affiliation: a Department of Food and Bioproduct Sciences ; University of Saskatchewan ; Saskatoon , Canada.|b Institute of Environment, Health and Societies; Brunel University; London , Uxbridge , United Kingdom.
authors.forename: Christopher H
authors.initials: CH
authors.lastname: Eskiw
mesh_terms: D000199:Actins / Q000378:metabolism; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects*; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D015850:Interleukin-6 / Q000378:metabolism; D016209:Interleukin-8 / Q000378:metabolism; D053605:Leukemia Inhibitory Factor / Q000378:metabolism; D000076222:Mechanistic Target of Rapamycin Complex 1; D046912:Multiprotein Complexes / Q000037:antagonists & inhibitors / Q000378:metabolism; D011401:Promoter Regions, Genetic; D050799:STAT5 Transcription Factor / Q000235:genetics / Q000378:metabolism*; D020123:Sirolimus / Q000494:pharmacology*; D058570:TOR Serine-Threonine Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism; D059467:Transcriptome; D025521:Tumor Suppressor Proteins / Q000235:genetics / Q000378:metabolism*; D015854:Up-Regulation / Q000187:drug effects
chemical_list: C526913:ACTC1 protein, human; D000199:Actins; D015850:Interleukin-6; D016209:Interleukin-8; D053605:Leukemia Inhibitory Factor; D046912:Multiprotein Complexes; D050799:STAT5 Transcription Factor; C494092:STAT5A protein, human; C494095:STAT5B protein, human; D025521:Tumor Suppressor Proteins; D000076222:Mechanistic Target of Rapamycin Complex 1; D058570:TOR Serine-Threonine Kinases; D020123:Sirolimus
keywords: RNA-seq; STAT5A/B; primary fibroblast; quiescence; rapamycin