BACKGROUND: Epigenetic modifications likely control the fate of hematopoietic stem cells (HSCs). The chromatin-modifying agents (CMAs), 5-aza-2'-deoxycytidine (5azaD) and trichostatin A (TSA), have previously been shown to expand HSCs from cord blood and marrow. Here we assessed whether CMA can also expand HSCs present in growth factor-mobilized human peripheral blood (MPB). STUDY DESIGN AND METHODS: 5azaD and TSA were sequentially added to CD34+ MPB cells in the presence of cytokines, and the cells were cultured for 9 days. RESULTS: After culture, a 3.6 ± 0.5-fold expansion of CD34+CD90+ cells, a 10.1 ± 0.5-fold expansion of primitive colony-forming unit (CFU)-mix, and a 2.2 ± 0.5-fold expansion of long-term cobblestone-area-forming cells (CAFCs) was observed in 5azaD/TSA-expanded cells. By contrast, cells cultured in cytokines without 5azaD/TSA displayed no expansion; rather, a reduction in CD34+CD90+ cells (0.7 ± 0.1-fold) and CAFCs (0.3 ± 0.1-fold) from their initial numbers was observed. Global hypomethylation corresponding with increased transcript levels of several genes implicated in HSC self-renewal, including HOXB4, GATA2, and EZH2, was observed in 5azaD/TSA-expanded MPB cells in contrast to controls. 5azaD/TSA-expanded MPB cells retained in vivo hematopoietic engraftment capacity. CONCLUSION: MPB CD34+ cells from donors can be expanded using 5azaD/TSA, and these expanded cells retain in vivo hematopoietic reconstitution capacity. This strategy may prove to be potentially useful to augment HSC numbers for patients who fail to mobilize.
BACKGROUND: Epigenetic modifications likely control the fate of hematopoietic stem cells (HSCs). The chromatin-modifying agents (CMAs), 5-aza-2'-deoxycytidine (5azaD) and trichostatin A (TSA), have previously been shown to expand HSCs from cord blood and marrow. Here we assessed whether CMA can also expand HSCs present in growth factor-mobilized human peripheral blood (MPB). STUDY DESIGN AND METHODS: 5azaD and TSA were sequentially added to CD34+ MPB cells in the presence of cytokines, and the cells were cultured for 9 days. RESULTS: After culture, a 3.6 ± 0.5-fold expansion of CD34+CD90+ cells, a 10.1 ± 0.5-fold expansion of primitive colony-forming unit (CFU)-mix, and a 2.2 ± 0.5-fold expansion of long-term cobblestone-area-forming cells (CAFCs) was observed in 5azaD/TSA-expanded cells. By contrast, cells cultured in cytokines without 5azaD/TSA displayed no expansion; rather, a reduction in CD34+CD90+ cells (0.7 ± 0.1-fold) and CAFCs (0.3 ± 0.1-fold) from their initial numbers was observed. Global hypomethylation corresponding with increased transcript levels of several genes implicated in HSC self-renewal, including HOXB4, GATA2, and EZH2, was observed in 5azaD/TSA-expanded MPB cells in contrast to controls. 5azaD/TSA-expanded MPB cells retained in vivo hematopoietic engraftment capacity. CONCLUSION: MPB CD34+ cells from donors can be expanded using 5azaD/TSA, and these expanded cells retain in vivo hematopoietic reconstitution capacity. This strategy may prove to be potentially useful to augment HSC numbers for patients who fail to mobilize.
Authors: M Lübbert; P Wijermans; R Kunzmann; G Verhoef; A Bosly; C Ravoet; M Andre; A Ferrant Journal: Br J Haematol Date: 2001-08 Impact factor: 6.998
Authors: Allen S Yang; Marcos R H Estécio; Ketan Doshi; Yutaka Kondo; Eloiza H Tajara; Jean-Pierre J Issa Journal: Nucleic Acids Res Date: 2004-02-18 Impact factor: 16.971
Authors: Roberta Pietrobono; Maria Grazia Pomponi; Elisabetta Tabolacci; Ben Oostra; Pietro Chiurazzi; Giovanni Neri Journal: Nucleic Acids Res Date: 2002-07-15 Impact factor: 16.971
Authors: Fiona McGregor; Alessandra Muntoni; Janis Fleming; Judith Brown; David H Felix; D Gordon MacDonald; E Kenneth Parkinson; Paul R Harrison Journal: Cancer Res Date: 2002-08-15 Impact factor: 12.701
Authors: Steven A Belinsky; Donna M Klinge; Christine A Stidley; Jean-Pierre Issa; James G Herman; Thomas H March; Stephen B Baylin Journal: Cancer Res Date: 2003-11-01 Impact factor: 12.701
Authors: Hiro Tatetsu; Myriam Armant; Fei Wang; Chong Gao; Shikiko Ueno; Xi Tian; Alex Federation; Jun Qi; James Bradner; Daniel G Tenen; Li Chai Journal: Exp Hematol Date: 2019-06-28 Impact factor: 3.084