Notch-mediated expansion of cord blood progenitors: maintenance of transcriptional and epigenetic fidelity

Notch-mediated expansion of cord blood progenitors: maintenance of transcriptional and epigenetic fidelity

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Access through your institution Buy or subscribe Recent advances have been made towards successful generation of human hematopoietic stem and progenitor cells (HSPCs) derived from embryonic, induced pluripotent or hematopoietic stem cells. However, whether these stem cell populations, which are generated after _ex vivo_ manipulation, are suitable for clinical application depends on the faithful preservation of their transcriptional, epigenetic and functional properties with respect to their primary cell counterparts. To overcome the significant delay in neutrophil recovery following cord blood (CB) transplantation (CBT), we developed methods for the _ex vivo_ expansion of CD34+ CB-derived HSPC by culture with the Notch-ligand Delta1. When infused into patients undergoing a myeloablative CBT, these cells have been shown to provide more rapid short-term myeloid reconstitution when co-infused with a non-manipulated CB unit(s). However, infusion of _ex vivo_ generated HSPCs for long-term hematopoietic reconstitution remains a goal in the context of stem cell transplantation and gene therapy, thus the development of methods to assess the safety of these cultured products are essential. This is a preview of subscription content, access via your institution ACCESS OPTIONS Access through your institution Subscribe to this journal Receive 12 print issues and online access $259.00 per year only $21.58 per issue Learn more Buy this article * Purchase on SpringerLink * Instant access to full article PDF Buy now Prices may be subject to local taxes which are calculated during checkout ADDITIONAL ACCESS OPTIONS: * Log in * Learn about institutional subscriptions * Read our FAQs * Contact customer support REFERENCES * Bock C, Kiskinis E, Verstappen G, Gu H, Boulting G, Smith ZD _et al_. Reference Maps of human ES and iPS cell variation enable high-throughput characterization of pluripotent cell lines. _Cell_ 2011; 144: 439–452. Article  CAS  Google Scholar  * Delaney C, Heimfeld S, Brashem-Stein C, Voorhies H, Manger RL, Bernstein ID . Notch-mediated expansion of human cord blood progenitor cells capable of rapid myeloid reconstitution. _Nat Med_ 2010; 16: 232–236. Article  CAS  Google Scholar  * Gentleman RC, Carey VJ, Bates DM, Bolstad B, Dettling M, Dudoit S _et al_. Bioconductor: open software development for computational biology and bioinformatics. _Genome Biol_ 2004; 5: R80. Article  Google Scholar  * Du P, Kibbe WA, Lin SM . Lumi: a pipeline for processing Illumina microarray. _Bioinformatics_ 2008; 24: 1547–1548. Article  CAS  Google Scholar  * Huang DW, Sherman BT, Lempicki RA . Systematic and integrative analysis of large gene lists using DAVID Bioinformatics Resources. _Nat Protoc_ 2009; 4: 44–57. Article  CAS  Google Scholar  * Huang DW, Sherman BT, Lempicki RA . Bioinformatics enrichment tools: paths toward the comprehensive functional analysis of large gene lists. _Nucleic Acids Res_ 2009; 37: 1–13. Article  Google Scholar  * Zhao M, Sun J, Zhao Z . TSGene: a web resource for tumor suppressor genes. _Nucleic Acids Res_ 2013; 41: D970–D976. Article  CAS  Google Scholar  * Rohrs S, Dirks WG, Meyer C, Marschalek R, Scherr M, Slany R _et al_. Hypomethylation and expression of BEX2, IGSF4, TIMP3 indicative of MLL translocations in acute myeloid leukemia. _Mol Cancer_ 2009; 8: 86. Article  Google Scholar  * Naderi A, Liu J, Hughes-Davies L . BEX2 has a functional interplay with c-Jun/JNK and p65/RelA in breast cancer. _Mol Cancer_ 2010; 9: 111. Article  Google Scholar  * Meissner A, Gnirke A, Bell GW, Ramsahoye B, Lander ES, Jaenisch R . Reduced representation bisulfite sequencing for comparative high-resolution DNA methylation analysis. _Nucleic Acids Res_ 2005; 33: 5868–5877. Article  CAS  Google Scholar  * Meissner A, Mikkelsen TS, Gu H, Wernig M, Hanna J, Sivachenko A _et al_. Genome-scale DNA methylation maps of pluripotent and differentiated cells. _Nature_ 2008; 454: 766–770. Article  CAS  Google Scholar  * Doi A, Park I-H, Wen B, Murakami P, Aryee MJ, Irizarry R _et al_. Differential methylation of tissue- and cancer-specific CpG island shores distinguishes human induced pluripotent stem cells, embryonic stem cells and fibroblasts. _Nat Genet_ 2009; 41: 1350–1353. Article  CAS  Google Scholar  * Laurent L, Ulitsky I, Slavin I, Tran H, Schork A, Morey R _et al_. Dynamic changes in the copy number of pluripotency and cell proliferation genes in human ESCs and iPSCs during reprogramming and time in culture. _Cell Stem Cell_ 2011; 8: 106–118. Article  CAS  Google Scholar  Download references ACKNOWLEDGEMENTS This work was supported by National Heart, Lung and Blood Institute grant U01HL100395 and National Institutes of Health Ruth L. Kirschstein National Research Service Award T32CA009351 (AD) and K12CA076930 (AD), as well as Hyundai Hope on Wheels 2013 Hope Grant (AD). CD is a Damon Runyon Clinical Investigator. AUTHOR INFORMATION AUTHORS AND AFFILIATIONS * Clinical Research Division, Pediatric Oncology, Fred Hutchinson Cancer Research Center, Seattle, WA, USA A Dahlberg, C Delaney & I D Bernstein * Department of Pediatrics, University of Washington, Seattle, Washington, USA A Dahlberg, C Delaney & I D Bernstein * Vaccine and Infectious Disease Division, PS Statistics, Fred Hutchinson Cancer Research Center, Seattle, WA, USA S Woo & R Gottardo * Broad Institute of MIT and Harvard, Cambridge, MA, USA P Boyle, A Gnirke, C Bock, B E Bernstein & A Meissner * Howard Hughes Medical Institute, Chevy Chase, MD, USA B E Bernstein * Department of Pathology, Massachusetts General Hospital and Harvard Medical School, Boston, MA, USA B E Bernstein Authors * A Dahlberg View author publications You can also search for this author inPubMed Google Scholar * S Woo View author publications You can also search for this author inPubMed Google Scholar * C Delaney View author publications You can also search for this author inPubMed Google Scholar * P Boyle View author publications You can also search for this author inPubMed Google Scholar * A Gnirke View author publications You can also search for this author inPubMed Google Scholar * C Bock View author publications You can also search for this author inPubMed Google Scholar * B E Bernstein View author publications You can also search for this author inPubMed Google Scholar * A Meissner View author publications You can also search for this author inPubMed Google Scholar * R Gottardo View author publications You can also search for this author inPubMed Google Scholar * I D Bernstein View author publications You can also search for this author inPubMed Google Scholar CORRESPONDING AUTHOR Correspondence to I D Bernstein. ETHICS DECLARATIONS COMPETING INTERESTS The Fred Hutchinson Cancer Research Center holds a patent on ‘methods for immortalizing cells’ that covers the use of Notch ligand for expansion of hematopoietic stem cells. IDB is an inventor on this patent. The remaining authors declare no conflict of interest. RIGHTS AND PERMISSIONS Reprints and permissions ABOUT THIS ARTICLE CITE THIS ARTICLE Dahlberg, A., Woo, S., Delaney, C. _et al._ Notch-mediated expansion of cord blood progenitors: maintenance of transcriptional and epigenetic fidelity. _Leukemia_ 29, 1948–1951 (2015). https://doi.org/10.1038/leu.2015.61 Download citation * Published: 06 March 2015 * Issue Date: September 2015 * DOI: https://doi.org/10.1038/leu.2015.61 SHARE THIS ARTICLE Anyone you share the following link with will be able to read this content: Get shareable link Sorry, a shareable link is not currently available for this article. Copy to clipboard Provided by the Springer Nature SharedIt content-sharing initiative

Access through your institution Buy or subscribe Recent advances have been made towards successful generation of human hematopoietic stem and progenitor cells (HSPCs) derived from embryonic,


induced pluripotent or hematopoietic stem cells. However, whether these stem cell populations, which are generated after _ex vivo_ manipulation, are suitable for clinical application


depends on the faithful preservation of their transcriptional, epigenetic and functional properties with respect to their primary cell counterparts. To overcome the significant delay in


neutrophil recovery following cord blood (CB) transplantation (CBT), we developed methods for the _ex vivo_ expansion of CD34+ CB-derived HSPC by culture with the Notch-ligand Delta1. When


infused into patients undergoing a myeloablative CBT, these cells have been shown to provide more rapid short-term myeloid reconstitution when co-infused with a non-manipulated CB unit(s).


However, infusion of _ex vivo_ generated HSPCs for long-term hematopoietic reconstitution remains a goal in the context of stem cell transplantation and gene therapy, thus the development of


methods to assess the safety of these cultured products are essential. This is a preview of subscription content, access via your institution ACCESS OPTIONS Access through your institution


Subscribe to this journal Receive 12 print issues and online access $259.00 per year only $21.58 per issue Learn more Buy this article * Purchase on SpringerLink * Instant access to full


article PDF Buy now Prices may be subject to local taxes which are calculated during checkout ADDITIONAL ACCESS OPTIONS: * Log in * Learn about institutional subscriptions * Read our FAQs *


Contact customer support REFERENCES * Bock C, Kiskinis E, Verstappen G, Gu H, Boulting G, Smith ZD _et al_. Reference Maps of human ES and iPS cell variation enable high-throughput


characterization of pluripotent cell lines. _Cell_ 2011; 144: 439–452. Article  CAS  Google Scholar  * Delaney C, Heimfeld S, Brashem-Stein C, Voorhies H, Manger RL, Bernstein ID .


Notch-mediated expansion of human cord blood progenitor cells capable of rapid myeloid reconstitution. _Nat Med_ 2010; 16: 232–236. Article  CAS  Google Scholar  * Gentleman RC, Carey VJ,


Bates DM, Bolstad B, Dettling M, Dudoit S _et al_. Bioconductor: open software development for computational biology and bioinformatics. _Genome Biol_ 2004; 5: R80. Article  Google Scholar 


* Du P, Kibbe WA, Lin SM . Lumi: a pipeline for processing Illumina microarray. _Bioinformatics_ 2008; 24: 1547–1548. Article  CAS  Google Scholar  * Huang DW, Sherman BT, Lempicki RA .


Systematic and integrative analysis of large gene lists using DAVID Bioinformatics Resources. _Nat Protoc_ 2009; 4: 44–57. Article  CAS  Google Scholar  * Huang DW, Sherman BT, Lempicki RA .


Bioinformatics enrichment tools: paths toward the comprehensive functional analysis of large gene lists. _Nucleic Acids Res_ 2009; 37: 1–13. Article  Google Scholar  * Zhao M, Sun J, Zhao Z


. TSGene: a web resource for tumor suppressor genes. _Nucleic Acids Res_ 2013; 41: D970–D976. Article  CAS  Google Scholar  * Rohrs S, Dirks WG, Meyer C, Marschalek R, Scherr M, Slany R _et


al_. Hypomethylation and expression of BEX2, IGSF4, TIMP3 indicative of MLL translocations in acute myeloid leukemia. _Mol Cancer_ 2009; 8: 86. Article  Google Scholar  * Naderi A, Liu J,


Hughes-Davies L . BEX2 has a functional interplay with c-Jun/JNK and p65/RelA in breast cancer. _Mol Cancer_ 2010; 9: 111. Article  Google Scholar  * Meissner A, Gnirke A, Bell GW, Ramsahoye


B, Lander ES, Jaenisch R . Reduced representation bisulfite sequencing for comparative high-resolution DNA methylation analysis. _Nucleic Acids Res_ 2005; 33: 5868–5877. Article  CAS 


Google Scholar  * Meissner A, Mikkelsen TS, Gu H, Wernig M, Hanna J, Sivachenko A _et al_. Genome-scale DNA methylation maps of pluripotent and differentiated cells. _Nature_ 2008; 454:


766–770. Article  CAS  Google Scholar  * Doi A, Park I-H, Wen B, Murakami P, Aryee MJ, Irizarry R _et al_. Differential methylation of tissue- and cancer-specific CpG island shores


distinguishes human induced pluripotent stem cells, embryonic stem cells and fibroblasts. _Nat Genet_ 2009; 41: 1350–1353. Article  CAS  Google Scholar  * Laurent L, Ulitsky I, Slavin I,


Tran H, Schork A, Morey R _et al_. Dynamic changes in the copy number of pluripotency and cell proliferation genes in human ESCs and iPSCs during reprogramming and time in culture. _Cell


Stem Cell_ 2011; 8: 106–118. Article  CAS  Google Scholar  Download references ACKNOWLEDGEMENTS This work was supported by National Heart, Lung and Blood Institute grant U01HL100395 and


National Institutes of Health Ruth L. Kirschstein National Research Service Award T32CA009351 (AD) and K12CA076930 (AD), as well as Hyundai Hope on Wheels 2013 Hope Grant (AD). CD is a Damon


Runyon Clinical Investigator. AUTHOR INFORMATION AUTHORS AND AFFILIATIONS * Clinical Research Division, Pediatric Oncology, Fred Hutchinson Cancer Research Center, Seattle, WA, USA A


Dahlberg, C Delaney & I D Bernstein * Department of Pediatrics, University of Washington, Seattle, Washington, USA A Dahlberg, C Delaney & I D Bernstein * Vaccine and Infectious


Disease Division, PS Statistics, Fred Hutchinson Cancer Research Center, Seattle, WA, USA S Woo & R Gottardo * Broad Institute of MIT and Harvard, Cambridge, MA, USA P Boyle, A Gnirke, C


Bock, B E Bernstein & A Meissner * Howard Hughes Medical Institute, Chevy Chase, MD, USA B E Bernstein * Department of Pathology, Massachusetts General Hospital and Harvard Medical


School, Boston, MA, USA B E Bernstein Authors * A Dahlberg View author publications You can also search for this author inPubMed Google Scholar * S Woo View author publications You can also


search for this author inPubMed Google Scholar * C Delaney View author publications You can also search for this author inPubMed Google Scholar * P Boyle View author publications You can


also search for this author inPubMed Google Scholar * A Gnirke View author publications You can also search for this author inPubMed Google Scholar * C Bock View author publications You can


also search for this author inPubMed Google Scholar * B E Bernstein View author publications You can also search for this author inPubMed Google Scholar * A Meissner View author publications


You can also search for this author inPubMed Google Scholar * R Gottardo View author publications You can also search for this author inPubMed Google Scholar * I D Bernstein View author


publications You can also search for this author inPubMed Google Scholar CORRESPONDING AUTHOR Correspondence to I D Bernstein. ETHICS DECLARATIONS COMPETING INTERESTS The Fred Hutchinson


Cancer Research Center holds a patent on ‘methods for immortalizing cells’ that covers the use of Notch ligand for expansion of hematopoietic stem cells. IDB is an inventor on this patent.


The remaining authors declare no conflict of interest. RIGHTS AND PERMISSIONS Reprints and permissions ABOUT THIS ARTICLE CITE THIS ARTICLE Dahlberg, A., Woo, S., Delaney, C. _et al._


Notch-mediated expansion of cord blood progenitors: maintenance of transcriptional and epigenetic fidelity. _Leukemia_ 29, 1948–1951 (2015). https://doi.org/10.1038/leu.2015.61 Download


citation * Published: 06 March 2015 * Issue Date: September 2015 * DOI: https://doi.org/10.1038/leu.2015.61 SHARE THIS ARTICLE Anyone you share the following link with will be able to read


this content: Get shareable link Sorry, a shareable link is not currently available for this article. Copy to clipboard Provided by the Springer Nature SharedIt content-sharing initiative