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Characterization of Transcription Factor Networks Involved in Umbilical Cord Blood CD34(+) Stem Cells-Derived Erythropoiesis
Fetal stem cells isolated from umbilical cord blood (UCB) possess a great capacity for proliferation and differentiation and serve as a valuable model system to study gene regulation. Expanded knowledge of the molecular control of hemoglobin synthesis will provide a basis for rational design of ther...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2014
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4161396/ https://www.ncbi.nlm.nih.gov/pubmed/25211130 http://dx.doi.org/10.1371/journal.pone.0107133 |
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author | Li, Biaoru Ding, Lianghao Yang, Chinrang Kang, Baolin Liu, Li Story, Michael D. Pace, Betty S. |
author_facet | Li, Biaoru Ding, Lianghao Yang, Chinrang Kang, Baolin Liu, Li Story, Michael D. Pace, Betty S. |
author_sort | Li, Biaoru |
collection | PubMed |
description | Fetal stem cells isolated from umbilical cord blood (UCB) possess a great capacity for proliferation and differentiation and serve as a valuable model system to study gene regulation. Expanded knowledge of the molecular control of hemoglobin synthesis will provide a basis for rational design of therapies for β-hemoglobinopathies. Transcriptome data are available for erythroid progenitors derived from adult stem cells, however studies to define molecular mechanisms controlling globin gene regulation during fetal erythropoiesis are limited. Here, we utilize UCB-CD34(+) stem cells induced to undergo erythroid differentiation to characterize the transcriptome and transcription factor networks (TFNs) associated with the γ/β-globin switch during fetal erythropoiesis. UCB-CD34(+) stem cells grown in the one-phase liquid culture system displayed a higher proliferative capacity than adult CD34(+) stem cells. The γ/β-globin switch was observed after day 42 during fetal erythropoiesis in contrast to adult progenitors where the switch occurred around day 21. To gain insights into transcription factors involved in globin gene regulation, microarray analysis was performed on RNA isolated from UCB-CD34(+) cell-derived erythroid progenitors harvested on day 21, 42, 49 and 56 using the HumanHT-12 Expression BeadChip. After data normalization, Gene Set Enrichment Analysis identified transcription factors (TFs) with significant changes in expression during the γ/β-globin switch. Forty-five TFs were silenced by day 56 (Profile-1) and 30 TFs were activated by day 56 (Profile-2). Both GSEA datasets were analyzed using the MIMI Cytoscape platform, which discovered TFNs centered on KLF4 and GATA2 (Profile-1) and KLF1 and GATA1 for Profile-2 genes. Subsequent shRNA studies in KU812 leukemia cells and human erythroid progenitors generated from UCB-CD34(+) cells supported a negative role of MAFB in γ-globin regulation. The characteristics of erythroblasts derived from UCB-CD34(+) stem cells including prolonged γ-globin expression combined with unique TFNs support novel mechanisms controlling the γ/β-globin switch during UCB-derived erythropoiesis. |
format | Online Article Text |
id | pubmed-4161396 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-41613962014-09-17 Characterization of Transcription Factor Networks Involved in Umbilical Cord Blood CD34(+) Stem Cells-Derived Erythropoiesis Li, Biaoru Ding, Lianghao Yang, Chinrang Kang, Baolin Liu, Li Story, Michael D. Pace, Betty S. PLoS One Research Article Fetal stem cells isolated from umbilical cord blood (UCB) possess a great capacity for proliferation and differentiation and serve as a valuable model system to study gene regulation. Expanded knowledge of the molecular control of hemoglobin synthesis will provide a basis for rational design of therapies for β-hemoglobinopathies. Transcriptome data are available for erythroid progenitors derived from adult stem cells, however studies to define molecular mechanisms controlling globin gene regulation during fetal erythropoiesis are limited. Here, we utilize UCB-CD34(+) stem cells induced to undergo erythroid differentiation to characterize the transcriptome and transcription factor networks (TFNs) associated with the γ/β-globin switch during fetal erythropoiesis. UCB-CD34(+) stem cells grown in the one-phase liquid culture system displayed a higher proliferative capacity than adult CD34(+) stem cells. The γ/β-globin switch was observed after day 42 during fetal erythropoiesis in contrast to adult progenitors where the switch occurred around day 21. To gain insights into transcription factors involved in globin gene regulation, microarray analysis was performed on RNA isolated from UCB-CD34(+) cell-derived erythroid progenitors harvested on day 21, 42, 49 and 56 using the HumanHT-12 Expression BeadChip. After data normalization, Gene Set Enrichment Analysis identified transcription factors (TFs) with significant changes in expression during the γ/β-globin switch. Forty-five TFs were silenced by day 56 (Profile-1) and 30 TFs were activated by day 56 (Profile-2). Both GSEA datasets were analyzed using the MIMI Cytoscape platform, which discovered TFNs centered on KLF4 and GATA2 (Profile-1) and KLF1 and GATA1 for Profile-2 genes. Subsequent shRNA studies in KU812 leukemia cells and human erythroid progenitors generated from UCB-CD34(+) cells supported a negative role of MAFB in γ-globin regulation. The characteristics of erythroblasts derived from UCB-CD34(+) stem cells including prolonged γ-globin expression combined with unique TFNs support novel mechanisms controlling the γ/β-globin switch during UCB-derived erythropoiesis. Public Library of Science 2014-09-11 /pmc/articles/PMC4161396/ /pubmed/25211130 http://dx.doi.org/10.1371/journal.pone.0107133 Text en © 2014 Li et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited. |
spellingShingle | Research Article Li, Biaoru Ding, Lianghao Yang, Chinrang Kang, Baolin Liu, Li Story, Michael D. Pace, Betty S. Characterization of Transcription Factor Networks Involved in Umbilical Cord Blood CD34(+) Stem Cells-Derived Erythropoiesis |
title | Characterization of Transcription Factor Networks Involved in Umbilical Cord Blood CD34(+) Stem Cells-Derived Erythropoiesis |
title_full | Characterization of Transcription Factor Networks Involved in Umbilical Cord Blood CD34(+) Stem Cells-Derived Erythropoiesis |
title_fullStr | Characterization of Transcription Factor Networks Involved in Umbilical Cord Blood CD34(+) Stem Cells-Derived Erythropoiesis |
title_full_unstemmed | Characterization of Transcription Factor Networks Involved in Umbilical Cord Blood CD34(+) Stem Cells-Derived Erythropoiesis |
title_short | Characterization of Transcription Factor Networks Involved in Umbilical Cord Blood CD34(+) Stem Cells-Derived Erythropoiesis |
title_sort | characterization of transcription factor networks involved in umbilical cord blood cd34(+) stem cells-derived erythropoiesis |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4161396/ https://www.ncbi.nlm.nih.gov/pubmed/25211130 http://dx.doi.org/10.1371/journal.pone.0107133 |
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