Silencing of ASXL1 impairs the granulomonocytic lineage potential of human CD34+ progenitor cells
The ASXL1 gene encodes a chromatin-binding protein involved in epigenetic regulation in haematopoietic cells. Loss-of-function ASXL1 mutations occur in patients with a range of myeloid malignancies and are associated with adverse outcome. We have used lentiviral-based shRNA technology to investigate...
| Main Authors: | , , , , , , , , , |
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| Format: | Journal article |
| Language: | English |
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2013
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| _version_ | 1826270394288439296 |
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| author | Davies, C Yip, B Fernandez-Mercado, M Woll, P Agirre, X Prosper, F Jacobsen, SE Wainscoat, J Pellagatti, A Boultwood, J |
| author_facet | Davies, C Yip, B Fernandez-Mercado, M Woll, P Agirre, X Prosper, F Jacobsen, SE Wainscoat, J Pellagatti, A Boultwood, J |
| author_sort | Davies, C |
| collection | OXFORD |
| description | The ASXL1 gene encodes a chromatin-binding protein involved in epigenetic regulation in haematopoietic cells. Loss-of-function ASXL1 mutations occur in patients with a range of myeloid malignancies and are associated with adverse outcome. We have used lentiviral-based shRNA technology to investigate the effects of ASXL1 silencing on cell proliferation, apoptosis, myeloid differentiation and global gene expression in human CD34+ cells differentiated along the myeloid lineage in vitro. ASXL1-deficient cells showed a significant decrease in the generation of CD11b+ and CD15+ cells, implicating impaired granulomonocytic differentiation. Furthermore, colony-forming assays showed a significant increase in the number of multipotent mixed lineage colony-forming unit (CFU-GEMM) colonies and a significant decrease in the numbers of granulocyte-macrophage CFU (CFU-GM) and granulocyte CFU (CFU-G) colonies in ASXL1-deficient cells. Our data suggests that ASXL1 knockdown perturbs human granulomonocytic differentiation. Gene expression profiling identified many deregulated genes in the ASXL1-deficient cells differentiated along the granulomonocytic lineage, and pathway analysis showed that the most significantly deregulated pathway was the LXR/RXR activation pathway. ASXL1 may play a key role in recruiting the polycomb repressor complex 2 (PRC2) to specific loci, and we found over-representation of PRC2 targets among the deregulated genes in ASXL1-deficient cells. These findings shed light on the functional role of ASXL1 in human myeloid differentiation. © 2013 Blackwell Publishing Ltd. |
| first_indexed | 2024-03-06T21:40:08Z |
| format | Journal article |
| id | oxford-uuid:47a1a092-52a8-4274-aeda-91123817d2dd |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-06T21:40:08Z |
| publishDate | 2013 |
| record_format | dspace |
| spelling | oxford-uuid:47a1a092-52a8-4274-aeda-91123817d2dd2022-03-26T15:21:10ZSilencing of ASXL1 impairs the granulomonocytic lineage potential of human CD34+ progenitor cellsJournal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:47a1a092-52a8-4274-aeda-91123817d2ddEnglishSymplectic Elements at Oxford2013Davies, CYip, BFernandez-Mercado, MWoll, PAgirre, XProsper, FJacobsen, SEWainscoat, JPellagatti, ABoultwood, JThe ASXL1 gene encodes a chromatin-binding protein involved in epigenetic regulation in haematopoietic cells. Loss-of-function ASXL1 mutations occur in patients with a range of myeloid malignancies and are associated with adverse outcome. We have used lentiviral-based shRNA technology to investigate the effects of ASXL1 silencing on cell proliferation, apoptosis, myeloid differentiation and global gene expression in human CD34+ cells differentiated along the myeloid lineage in vitro. ASXL1-deficient cells showed a significant decrease in the generation of CD11b+ and CD15+ cells, implicating impaired granulomonocytic differentiation. Furthermore, colony-forming assays showed a significant increase in the number of multipotent mixed lineage colony-forming unit (CFU-GEMM) colonies and a significant decrease in the numbers of granulocyte-macrophage CFU (CFU-GM) and granulocyte CFU (CFU-G) colonies in ASXL1-deficient cells. Our data suggests that ASXL1 knockdown perturbs human granulomonocytic differentiation. Gene expression profiling identified many deregulated genes in the ASXL1-deficient cells differentiated along the granulomonocytic lineage, and pathway analysis showed that the most significantly deregulated pathway was the LXR/RXR activation pathway. ASXL1 may play a key role in recruiting the polycomb repressor complex 2 (PRC2) to specific loci, and we found over-representation of PRC2 targets among the deregulated genes in ASXL1-deficient cells. These findings shed light on the functional role of ASXL1 in human myeloid differentiation. © 2013 Blackwell Publishing Ltd. |
| spellingShingle | Davies, C Yip, B Fernandez-Mercado, M Woll, P Agirre, X Prosper, F Jacobsen, SE Wainscoat, J Pellagatti, A Boultwood, J Silencing of ASXL1 impairs the granulomonocytic lineage potential of human CD34+ progenitor cells |
| title | Silencing of ASXL1 impairs the granulomonocytic lineage potential of human CD34+ progenitor cells |
| title_full | Silencing of ASXL1 impairs the granulomonocytic lineage potential of human CD34+ progenitor cells |
| title_fullStr | Silencing of ASXL1 impairs the granulomonocytic lineage potential of human CD34+ progenitor cells |
| title_full_unstemmed | Silencing of ASXL1 impairs the granulomonocytic lineage potential of human CD34+ progenitor cells |
| title_short | Silencing of ASXL1 impairs the granulomonocytic lineage potential of human CD34+ progenitor cells |
| title_sort | silencing of asxl1 impairs the granulomonocytic lineage potential of human cd34 progenitor cells |
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