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Silencing of ASXL 1 impairs the granulomonocytic lineage potential of human CD 34 + progenitor cells
Author(s) -
Davies Carwyn,
Yip Bon Ham,
FernandezMercado Marta,
Woll Petter S.,
Agirre Xabier,
Prosper Felipe,
Jacobsen Sten E.,
Wainscoat James S.,
Pellagatti Andrea,
Boultwood Jacqueline
Publication year - 2013
Publication title -
british journal of haematology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.907
H-Index - 186
eISSN - 1365-2141
pISSN - 0007-1048
DOI - 10.1111/bjh.12217
Subject(s) - gene silencing , progenitor cell , cd34 , progenitor , lineage (genetic) , biology , virology , stem cell , microbiology and biotechnology , genetics , gene
Summary The ASXL 1 gene encodes a chromatin‐binding protein involved in epigenetic regulation in haematopoietic cells. Loss‐of‐function ASXL 1 mutations occur in patients with a range of myeloid malignancies and are associated with adverse outcome. We have used lentiviral‐based sh RNA technology to investigate the effects of ASXL 1 silencing on cell proliferation, apoptosis, myeloid differentiation and global gene expression in human CD 34 + cells differentiated along the myeloid lineage in vitro . ASXL 1‐deficient cells showed a significant decrease in the generation of CD 11b + and CD 15 + 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 ASXL 1‐deficient cells. Our data suggests that ASXL 1 knockdown perturbs human granulomonocytic differentiation. Gene expression profiling identified many deregulated genes in the ASXL 1‐deficient cells differentiated along the granulomonocytic lineage, and pathway analysis showed that the most significantly deregulated pathway was the LXR / RXR activation pathway. ASXL 1 may play a key role in recruiting the polycomb repressor complex 2 ( PRC 2) to specific loci, and we found over‐representation of PRC 2 targets among the deregulated genes in ASXL 1‐deficient cells. These findings shed light on the functional role of ASXL 1 in human myeloid differentiation.