Decitabine Suspends Human CD34+ Cell Differentiation and Proliferation during Lentiviral Transduction

Efficient ex vivo transduction of hematopoietic stem cells (HSCs) is encumbered by differentiation which reduces engraftment. We hypothesized that inhibiting DNA methyltransferase with decitabine would block differentiation of transduced CD34+ cells under cytokine stimulation and thus improve transd...

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Main Authors:	Uchida, Naoya, Hsieh, Matthew M., Platner, Charlotte, Saunthararajah, Yogen, Tisdale, John F.
Format:	Online
Language:	English
Published:	Public Library of Science 2014
Online Access:	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4121241/

id	pubmed-4121241
recordtype	oai_dc
spelling	pubmed-41212412014-08-05 Decitabine Suspends Human CD34+ Cell Differentiation and Proliferation during Lentiviral Transduction Uchida, Naoya Hsieh, Matthew M. Platner, Charlotte Saunthararajah, Yogen Tisdale, John F. Research Article Efficient ex vivo transduction of hematopoietic stem cells (HSCs) is encumbered by differentiation which reduces engraftment. We hypothesized that inhibiting DNA methyltransferase with decitabine would block differentiation of transduced CD34+ cells under cytokine stimulation and thus improve transduction efficiency for engrafting HSCs. Human CD34+ cells in cytokine-containing media were treated with or without decitabine for 24 or 48 hours, and then these cells were transduced with a GFP-expressing lentiviral vector. Utilizing decitabine pre-treatment for 48 hours, we observed an equivalent percentage of successfully transduced cells (GFP-positivity) and a higher percentage of cells that retained CD34 positivity, compared to no decitabine exposure. Cell proliferation was inhibited after decitabine exposure. Similar results were observed among CD34+ cells from six different donors. Repopulating activity was evaluated by transplantation into NOD/SCID/IL2Rγnull mice and demonstrated an equivalent percentage of GFP-positivity in human cells from decitabine-treated samples and a trend for higher human cell engraftment (measured 20–24 weeks after transplantation), compared to no decitabine exposure. In conclusion, ex vivo decitabine exposure inhibits both differentiation and proliferation in transduced human CD34+ cells and modestly increases the engraftment ability in xenograft mice, while the transduction efficiency is equivalent in decitabine exposure, suggesting improvement of lentiviral transduction for HSCs. Public Library of Science 2014-08-04 /pmc/articles/PMC4121241/ /pubmed/25089909 http://dx.doi.org/10.1371/journal.pone.0104022 Text en https://creativecommons.org/publicdomain/zero/1.0/ This is an open-access article distributed under the terms of the Creative Commons Public Domain declaration, which stipulates that, once placed in the public domain, this work may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose.
repository_type	Open Access Journal
institution_category	Foreign Institution
institution	US National Center for Biotechnology Information
building	NCBI PubMed
collection	Online Access
language	English
format	Online
author	Uchida, Naoya Hsieh, Matthew M. Platner, Charlotte Saunthararajah, Yogen Tisdale, John F.
spellingShingle	Uchida, Naoya Hsieh, Matthew M. Platner, Charlotte Saunthararajah, Yogen Tisdale, John F. Decitabine Suspends Human CD34+ Cell Differentiation and Proliferation during Lentiviral Transduction
author_facet	Uchida, Naoya Hsieh, Matthew M. Platner, Charlotte Saunthararajah, Yogen Tisdale, John F.
author_sort	Uchida, Naoya
title	Decitabine Suspends Human CD34+ Cell Differentiation and Proliferation during Lentiviral Transduction
title_short	Decitabine Suspends Human CD34+ Cell Differentiation and Proliferation during Lentiviral Transduction
title_full	Decitabine Suspends Human CD34+ Cell Differentiation and Proliferation during Lentiviral Transduction
title_fullStr	Decitabine Suspends Human CD34+ Cell Differentiation and Proliferation during Lentiviral Transduction
title_full_unstemmed	Decitabine Suspends Human CD34+ Cell Differentiation and Proliferation during Lentiviral Transduction
title_sort	decitabine suspends human cd34+ cell differentiation and proliferation during lentiviral transduction
description	Efficient ex vivo transduction of hematopoietic stem cells (HSCs) is encumbered by differentiation which reduces engraftment. We hypothesized that inhibiting DNA methyltransferase with decitabine would block differentiation of transduced CD34+ cells under cytokine stimulation and thus improve transduction efficiency for engrafting HSCs. Human CD34+ cells in cytokine-containing media were treated with or without decitabine for 24 or 48 hours, and then these cells were transduced with a GFP-expressing lentiviral vector. Utilizing decitabine pre-treatment for 48 hours, we observed an equivalent percentage of successfully transduced cells (GFP-positivity) and a higher percentage of cells that retained CD34 positivity, compared to no decitabine exposure. Cell proliferation was inhibited after decitabine exposure. Similar results were observed among CD34+ cells from six different donors. Repopulating activity was evaluated by transplantation into NOD/SCID/IL2Rγnull mice and demonstrated an equivalent percentage of GFP-positivity in human cells from decitabine-treated samples and a trend for higher human cell engraftment (measured 20–24 weeks after transplantation), compared to no decitabine exposure. In conclusion, ex vivo decitabine exposure inhibits both differentiation and proliferation in transduced human CD34+ cells and modestly increases the engraftment ability in xenograft mice, while the transduction efficiency is equivalent in decitabine exposure, suggesting improvement of lentiviral transduction for HSCs.
publisher	Public Library of Science
publishDate	2014
url	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4121241/
_version_	1613120746880499712

Decitabine Suspends Human CD34+ Cell Differentiation and Proliferation during Lentiviral Transduction

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