title: Large-scale transcriptional profiling and functional assays reveal important roles for Rho-GTPase signalling and SCL during haematopoietic differentiation of human embryonic stem cells.
abstract: Understanding the transcriptional cues that direct differentiation of human embryonic stem cells (hESCs) and human-induced pluripotent stem cells to defined and functional cell types is essential for future clinical applications. In this study, we have compared transcriptional profiles of haematopoietic progenitors derived from hESCs at various developmental stages of a feeder- and serum-free differentiation method and show that the largest transcriptional changes occur during the first 4 days of differentiation. Data mining on the basis of molecular function revealed Rho-GTPase signalling as a key regulator of differentiation. Inhibition of this pathway resulted in a significant reduction in the numbers of emerging haematopoietic progenitors throughout the differentiation window, thereby uncovering a previously unappreciated role for Rho-GTPase signalling during human haematopoietic development. Our analysis indicated that SCL was the 11th most upregulated transcript during the first 4 days of the hESC differentiation process. Overexpression of SCL in hESCs promoted differentiation to meso-endodermal lineages, the emergence of haematopoietic and erythro-megakaryocytic progenitors and accelerated erythroid differentiation. Importantly, intrasplenic transplantation of SCL-overexpressing hESC-derived haematopoietic cells enhanced recovery from induced acute anaemia without significant cell engraftment, suggesting a paracrine-mediated effect.
authors.affiliation: Institute of Genetic Medicine, Newcastle University, Newcastle upon Tyne, UK.
authors.forename: Sun
authors.initials: S
authors.lastname: Yung
authors.forename: Maria
authors.initials: M
authors.lastname: Ledran
authors.forename: Inmaculada
authors.initials: I
authors.lastname: Moreno-Gimeno
authors.forename: Ana
authors.initials: A
authors.lastname: Conesa
authors.forename: David
authors.initials: D
authors.lastname: Montaner
authors.forename: Joaquín
authors.initials: J
authors.lastname: Dopazo
authors.forename: Ian
authors.initials: I
authors.lastname: Dimmick
authors.forename: Nicholas J
authors.initials: NJ
authors.lastname: Slater
authors.forename: Lamin
authors.initials: L
authors.lastname: Marenah
authors.forename: Pedro J
authors.initials: PJ
authors.lastname: Real
authors.forename: Iliana
authors.initials: I
authors.lastname: Paraskevopoulou
authors.forename: Viviana
authors.initials: V
authors.lastname: Bisbal
authors.forename: Deborah
authors.initials: D
authors.lastname: Burks
authors.forename: Mauro
authors.initials: M
authors.lastname: Santibanez-Koref
authors.forename: Ruben
authors.initials: R
authors.lastname: Moreno
authors.forename: Joanne
authors.initials: J
authors.lastname: Mountford
authors.forename: Pablo
authors.initials: P
authors.lastname: Menendez
authors.forename: Lyle
authors.initials: L
authors.lastname: Armstrong
authors.forename: Majlinda
authors.initials: M
authors.lastname: Lako
mesh_terms: D000208:Acute Disease; D000743:Anemia, Hemolytic / Q000235:genetics / Q000473:pathology / Q000628:therapy; D000818:Animals; D051792:Basic Helix-Loop-Helix Proteins / Q000235:genetics* / Q000378:metabolism; D002454:Cell Differentiation / Q000235:genetics*; D002460:Cell Line; D019070:Cell Lineage / Q000235:genetics; D016000:Cluster Analysis; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism; D041905:Erythroid Cells / Q000166:cytology / Q000378:metabolism; D005434:Flow Cytometry; D020869:Gene Expression Profiling; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000378:metabolism; D006801:Humans; D051379:Mice; D022423:Myeloid Cells / Q000166:cytology; D019899:Paracrine Communication / Q000235:genetics; D011518:Proto-Oncogene Proteins / Q000235:genetics* / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction / Q000235:genetics; D033581:Stem Cell Transplantation; D000073942:T-Cell Acute Lymphocytic Leukemia Protein 1; D059467:Transcriptome / Q000235:genetics*; D020741:rho GTP-Binding Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D051792:Basic Helix-Loop-Helix Proteins; D011518:Proto-Oncogene Proteins; D000073942:T-Cell Acute Lymphocytic Leukemia Protein 1; D020741:rho GTP-Binding Proteins; C067143:TAL1 protein, human