title: Preferential Propagation of Competent SIX2+ Nephronic Progenitors by LIF/ROCKi Treatment of the Metanephric Mesenchyme.
abstract: Understanding the mechanisms responsible for nephrogenic stem cell preservation and commitment is fundamental to harnessing the potential of the metanephric mesenchyme (MM) for nephron regeneration. Accordingly, we established a culture model that preferentially expands the MM SIX2+ progenitor pool using leukemia inhibitory factor (LIF), a Rho kinase inhibitor (ROCKi), and extracellular matrix. Passaged MM cells express the key stem cell regulators Six2 and Pax2 and remain competent to respond to WNT4 induction and form mature tubular epithelia and glomeruli. Mechanistically, LIF activates STAT, which binds to a Stat consensus sequence in the Six2 proximal promoter and sustains SIX2 levels. ROCKi, on the other hand, attenuates the LIF-induced differentiation activity of JNK. Concomitantly, the combination of LIF/ROCKi upregulates Slug expression and activates YAP, which maintains SIX2, PAX2, and SALL1. Using this novel model, our study underscores the pivotal roles of SIX2 and YAP in MM stem cell stability.
authors: National Cancer Institute-Frederick, Frederick, MD 21702, USA; Department of Kidney Development, Institute of Molecular Embryology and Genetics, Kumamoto University, 2-2-1 Honjo, Kumamoto 860-0811, Japan; Program for Leading Graduate Schools, Health Life Science: Interdisciplinary and Glocal (Global and Local) Oriented Program, Kumamoto University, 2-2-1 Honjo, Kumamoto 860-0811, Japan.
authors: Shunsuke
authors: S
authors: Tanigawa
authors: National Cancer Institute-Frederick, Frederick, MD 21702, USA.
authors: Nirmala
authors: N
authors: Sharma
authors: National Cancer Institute-Frederick, Frederick, MD 21702, USA.
authors: Michael D
authors: MD
authors: Hall
authors: Department of Kidney Development, Institute of Molecular Embryology and Genetics, Kumamoto University, 2-2-1 Honjo, Kumamoto 860-0811, Japan; Program for Leading Graduate Schools, Health Life Science: Interdisciplinary and Glocal (Global and Local) Oriented Program, Kumamoto University, 2-2-1 Honjo, Kumamoto 860-0811, Japan.
authors: Ryuichi
authors: R
authors: Nishinakamura
authors: National Cancer Institute-Frederick, Frederick, MD 21702, USA. Electronic address: perantoa@mail.nih.gov.
authors: Alan O
authors: AO
authors: Perantoni
mesh_terms: D000818:Animals; D018398:Homeodomain Proteins / Q000096:biosynthesis*; D053605:Leukemia Inhibitory Factor / Q000494:pharmacology*; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000378:metabolism*; D051379:Mice; D009399:Nephrons / Q000166:cytology / Q000378:metabolism*; D051762:PAX2 Transcription Factor / Q000096:biosynthesis; D047428:Protein Kinase Inhibitors / Q000494:pharmacology*; D000071250:Snail Family Transcription Factors; D014157:Transcription Factors / Q000096:biosynthesis*; D054460:rho-Associated Kinases / Q000037:antagonists & inhibitors*
chemical_list: D018398:Homeodomain Proteins; D053605:Leukemia Inhibitory Factor; C507838:Lif protein, mouse; D051762:PAX2 Transcription Factor; C497319:Pax2 protein, mouse; D047428:Protein Kinase Inhibitors; C491254:Sall1 protein, mouse; C497420:Six2 protein, mouse; C000604449:Snai2 protein, mouse; D000071250:Snail Family Transcription Factors; D014157:Transcription Factors; D054460:rho-Associated Kinases