title: Discovery of a Small-Molecule BMP Sensitizer for Human Embryonic Stem Cell Differentiation.
abstract: Sorely missing from the "toolkit" for directed differentiation of stem/progenitor cells are agonists of the BMP-signaling pathway. Using a high-throughput chemical screen, we discovered that PD407824, a checkpoint kinase 1 (CHK1) inhibitor, increases the sensitivity of cells to sub-threshold amounts of BMP4. We show utility of the compound in the directed differentiation of human embryonic stem cells toward mesoderm or cytotrophoblast stem cells. Blocking CHK1 activity using pharmacological compounds or CHK1 knockout using single guide RNA (sgRNA) confirmed that CHK1 inhibition increases the sensitivity to BMP4 treatment. Additional mechanistic studies indicate that CHK1 inhibition depletes p21 levels, thereby activating CDK8/9, which then phosphorylates the SMAD2/3 linker region, leading to decreased levels of SMAD2/3 protein and enhanced levels of nuclear SMAD1. This study provides insight into mechanisms controlling the BMP/transforming growth factor beta (TGF-β) signaling pathways and a useful pharmacological reagent for directed differentiation of stem cells.
authors.affiliation: Key Laboratory of Pesticide and Chemical Biology (CCNU), Ministry of Education, College of Chemistry, Central China Normal University, Wuhan, Hubei 430079, China; Department of Surgery, Weill Cornell Medical College, New York, NY 10065, USA.
authors.forename: Lingling
authors.initials: L
authors.lastname: Feng
authors.affiliation: Department of Surgery, Weill Cornell Medical College, New York, NY 10065, USA.
authors.forename: Brandoch
authors.initials: B
authors.lastname: Cook
authors.affiliation: Department of Surgery, Weill Cornell Medical College, New York, NY 10065, USA.
authors.forename: Su-Yi
authors.initials: SY
authors.lastname: Tsai
authors.affiliation: Department of Surgery, Weill Cornell Medical College, New York, NY 10065, USA.
authors.forename: Ting
authors.initials: T
authors.lastname: Zhou
authors.affiliation: Department of Surgery, Weill Cornell Medical College, New York, NY 10065, USA.
authors.forename: Brooke
authors.initials: B
authors.lastname: LaFlamme
authors.affiliation: Department of Surgery, Weill Cornell Medical College, New York, NY 10065, USA. Electronic address: tre2003@med.cornell.edu.
authors.forename: Todd
authors.initials: T
authors.lastname: Evans
authors.affiliation: Department of Surgery, Weill Cornell Medical College, New York, NY 10065, USA. Electronic address: shc2034@med.cornell.edu.
authors.forename: Shuibing
authors.initials: S
authors.lastname: Chen
mesh_terms: D000818:Animals; D055415:Bone Morphogenetic Protein 4 / Q000378:metabolism; D002227:Carbazoles / Q000737:chemistry / Q000494:pharmacology; D002454:Cell Differentiation / Q000187:drug effects; D019070:Cell Lineage / Q000187:drug effects; D065150:Cellular Reprogramming / Q000187:drug effects; D000071877:Checkpoint Kinase 1 / Q000378:metabolism; D000066449:Human Embryonic Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D008648:Mesoderm / Q000166:cytology; D051379:Mice; D008954:Models, Biological; D032446:Myoblasts / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D010006:Osteoblasts / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects; D051785:Smad Proteins / Q000378:metabolism; D054852:Small Molecule Libraries / Q000737:chemistry / Q000494:pharmacology*; D014327:Trophoblasts / Q000166:cytology / Q000187:drug effects
chemical_list: D055415:Bone Morphogenetic Protein 4; D002227:Carbazoles; C544549:PD 407824; D051785:Smad Proteins; D054852:Small Molecule Libraries; D000071877:Checkpoint Kinase 1