TY - JOUR T1 - Large-scale transcriptional profiling and functional assays reveal important roles for Rho-GTPase signalling and SCL during haematopoietic differentiation of human embryonic stem cells. JF - Hum Mol Genet Y1 - 2011 A1 - Yung, Sun A1 - Ledran, Maria A1 - Moreno-Gimeno, Inmaculada A1 - Conesa, Ana A1 - Montaner, David A1 - Dopazo, Joaquin A1 - Dimmick, Ian A1 - Slater, Nicholas J A1 - Marenah, Lamin A1 - Real, Pedro J A1 - Paraskevopoulou, Iliana A1 - Bisbal, Viviana A1 - Burks, Deborah A1 - Santibanez-Koref, Mauro A1 - Moreno, Ruben A1 - Mountford, Joanne A1 - Menendez, Pablo A1 - Armstrong, Lyle A1 - Lako, Majlinda KW - Acute Disease KW - Anemia, Hemolytic KW - Animals KW - Basic Helix-Loop-Helix Transcription Factors KW - Cell Differentiation KW - Cell Line KW - Cell Lineage KW - Cluster Analysis KW - Embryonic Stem Cells KW - Erythroid Cells KW - Flow Cytometry KW - Gene Expression Profiling KW - Hematopoietic Stem Cells KW - Humans KW - Mice KW - Myeloid Cells KW - Paracrine Communication KW - Proto-Oncogene Proteins KW - Reverse Transcriptase Polymerase Chain Reaction KW - rho GTP-Binding Proteins KW - Signal Transduction KW - Stem Cell Transplantation KW - T-Cell Acute Lymphocytic Leukemia Protein 1 KW - Transcriptome AB -

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.

VL - 20 IS - 24 U1 - https://www.ncbi.nlm.nih.gov/pubmed/21937587?dopt=Abstract ER -