title: Stroma-Derived Connective Tissue Growth Factor Maintains Cell Cycle Progression and Repopulation Activity of Hematopoietic Stem Cells In Vitro.
abstract: Hematopoietic stem cells (HSCs) are preserved in co-cultures with UG26-1B6 stromal cells or their conditioned medium. We performed a genome-wide study of gene expression changes of UG26-1B6 stromal cells in contact with Lineage⁻ SCA-1⁺ KIT⁺ (LSK) cells. This analysis identified connective tissue growth factor (CTGF) to be upregulated in response to LSK cells. We found that co-culture of HSCs on CTGF knockdown stroma (shCtgf) shows impaired engraftment and long-term quality. Further experiments demonstrated that CD34⁻ CD48⁻ CD150⁺ LSK (CD34⁻ SLAM) cell numbers from shCtgf co-cultures increase in G0 and senescence and show delayed time to first cell division. To understand this observation, a CTGF signaling network model was assembled, which was experimentally validated. In co-culture experiments of CD34⁻ SLAM cells with shCtgf stromal cells, we found that SMAD2/3-dependent signaling was activated, with increasing p27(Kip1) expression and downregulating cyclin D1. Our data support the view that LSK cells modulate gene expression in the niche to maintain repopulating HSC activity.
authors.affiliation: 3rd Department of Internal Medicine, Klinikum Rechts der Isar der Technischen Universität München, 81675 Munich, Germany.
authors.forename: Rouzanna
authors.initials: R
authors.lastname: Istvánffy
authors.affiliation: 3rd Department of Internal Medicine, Klinikum Rechts der Isar der Technischen Universität München, 81675 Munich, Germany; Helmholtz Zentrum München, Institute of Bioinformatics and Systems Biology, 85764 Neuherberg, Germany.
authors.forename: Baiba
authors.initials: B
authors.lastname: Vilne
authors.affiliation: 3rd Department of Internal Medicine, Klinikum Rechts der Isar der Technischen Universität München, 81675 Munich, Germany.
authors.forename: Christina
authors.initials: C
authors.lastname: Schreck
authors.affiliation: 3rd Department of Internal Medicine, Klinikum Rechts der Isar der Technischen Universität München, 81675 Munich, Germany.
authors.forename: Franziska
authors.initials: F
authors.lastname: Ruf
authors.affiliation: 3rd Department of Internal Medicine, Klinikum Rechts der Isar der Technischen Universität München, 81675 Munich, Germany.
authors.forename: Charlotta
authors.initials: C
authors.lastname: Pagel
authors.affiliation: 3rd Department of Internal Medicine, Klinikum Rechts der Isar der Technischen Universität München, 81675 Munich, Germany.
authors.forename: Sandra
authors.initials: S
authors.lastname: Grziwok
authors.affiliation: Institute of Medical Microbiology, Immunology, and Hygiene, Technische Universität München, 81675 Munich, Germany.
authors.forename: Lynette
authors.initials: L
authors.lastname: Henkel
authors.affiliation: Institute of Medical Microbiology, Immunology, and Hygiene, Technische Universität München, 81675 Munich, Germany.
authors.forename: Olivia
authors.initials: O
authors.lastname: Prazeres da Costa
authors.affiliation: Clinic for Radiooncology and Radiation Therapy, Klinikums Rechts der Isar der Technischen Universität München, 81675 Munich, Germany.
authors.forename: Johannes
authors.initials: J
authors.lastname: Berndt
authors.affiliation: Helmholtz Zentrum München, Institute of Bioinformatics and Systems Biology, 85764 Neuherberg, Germany.
authors.forename: Volker
authors.initials: V
authors.lastname: Stümpflen
authors.affiliation: 3rd Department of Internal Medicine, Klinikum Rechts der Isar der Technischen Universität München, 81675 Munich, Germany; German Consortium for Translational Research (DKTK), Partner Site Technical University Munich, 81675 Munich, Germany.
authors.forename: Katharina S
authors.initials: KS
authors.lastname: Götze
authors.affiliation: Institute of Medical Microbiology, Immunology, and Hygiene, Technische Universität München, 81675 Munich, Germany.
authors.forename: Matthias
authors.initials: M
authors.lastname: Schiemann
authors.affiliation: 3rd Department of Internal Medicine, Klinikum Rechts der Isar der Technischen Universität München, 81675 Munich, Germany; German Consortium for Translational Research (DKTK), Partner Site Technical University Munich, 81675 Munich, Germany.
authors.forename: Christian
authors.initials: C
authors.lastname: Peschel
authors.affiliation: Helmholtz Zentrum München, Institute of Bioinformatics and Systems Biology, 85764 Neuherberg, Germany; Genome-Oriented Bioinformatics, Technische Universität München, Weihenstephan, 85354 Freising, Germany.
authors.forename: Hans-Werner
authors.initials: HW
authors.lastname: Mewes
authors.affiliation: 3rd Department of Internal Medicine, Klinikum Rechts der Isar der Technischen Universität München, 81675 Munich, Germany. Electronic address: robert.oostendorp@tum.de.
authors.forename: Robert A J
authors.initials: RAJ
authors.lastname: Oostendorp
mesh_terms: D000818:Animals; D002453:Cell Cycle; D002460:Cell Line; D002478:Cells, Cultured; D055513:Connective Tissue Growth Factor / Q000378:metabolism / Q000494:pharmacology*; D019938:Cyclin D1 / Q000235:genetics / Q000378:metabolism; D050760:Cyclin-Dependent Kinase Inhibitor p27 / Q000235:genetics / Q000378:metabolism; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D051899:Smad2 Protein / Q000378:metabolism; D051900:Smad3 Protein / Q000378:metabolism; D055153:Stem Cell Niche; D017154:Stromal Cells / Q000378:metabolism*
chemical_list: C526318:CCN2 protein, mouse; D051899:Smad2 Protein; D051900:Smad3 Protein; D019938:Cyclin D1; D055513:Connective Tissue Growth Factor; D050760:Cyclin-Dependent Kinase Inhibitor p27