Pegylated granulocyte colony-stimulating factor mobilizes CD34+ cells with different stem and progenitor subsets and distinct functional properties in comparison with unconjugated granulocyte colony-stimulating factor

Ingmar Bruns, Ulrich G. Steidl, Johannes C. Fischer, Akos Czibere, Guido Kobbe, Sascha Raschke, Raminder Singh, Roland Fenk, Michael Rosskopf, Sabrina Pechtel, Arndt Von Haeseler, Peter Wernet, Daniel G. Tenen, Rainer Haas, Ralf Kronenwett

Research output: Contribution to journalArticle

31 Citations (Scopus)

Abstract

Background: Pegylated granulocyte colony-stimulating factor (G-CSF) has recently been introduced as a new compound for mobilization of CD34+ hematopoietic stem and progenitor cells. In this study, we compared the molecular and functional characteristics of CD34+ cells mobilized by pegylated G-CSF with those mobilized by unconjugated G-CSF. Design and Methods: Gene expression of immunomagnetically enriched CD34+ cells from leukapheresis products of patients who were given pegylated-G-CSF or unconjugated G-CSF was analyzed using Affymetrix HG Focus microarrays and quantitative reverse transcriptase polymerase chain reaction. Flow cytometry and fluorescence activated cell sorting was conducted to assess the CD34 + subset composition and to obtain Lin-, CD34+, CD38- hematopoietic stem cells. Cell cycle assays and clonogenic assays were performed for functional corroboration. Results: Pegylated G-CSF and unconjugated G-CSF mobilized CD34+ and hematopoietic stem cells with different molecular phenotypes and functional properties. The CD34+ cells mobilized by pegylated G-CSF had higher expression levels of genes indicative of early hematopoiesis, including HOXA9, MEIS1 and GATA3. We found lower expression of genes characteristic of erythroid and later stages of myeloid differentiation and a lower functional burst-forming unit erythroid/colony-forming unit-granulocyte-macrophage ratio. Consistently, greater numbers of hematopoietic stem cells and common myeloid progenitors and fewer megakaryocyte-erthrocyte progenitors were found in the pegylated-G-CSF-mobilized CD34+ cells. Additionally, sorted pegylated-G-CSF-mobilized hematopoietic stem cells displayed higher expression of HOXA9 in comparison to G-CSF-mobilized hematopoietic stem cells. In line with the gene expression data, CD34+ cells mobilized by pegylated G-CSF, as well as sorted hematopoietic stem cells, showed a significantly greater cell cycle activity. Conclusions: Stimulation with pegylated-G-CSF or G-CSF results in different expression of key regulatory genes and different functional properties of mobilized hematopoietic stem cells as well as their progeny, a finding that might be relevant for the application of these cells in blood stem cell transplantation.

Original languageEnglish (US)
Pages (from-to)347-355
Number of pages9
JournalHaematologica
Volume93
Issue number3
DOIs
StatePublished - Mar 2008

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Granulocyte Colony-Stimulating Factor
Hematopoietic Stem Cells
Gene Expression
Cell Cycle
Flow Cytometry
Megakaryocyte Progenitor Cells
Myeloid Progenitor Cells
Leukapheresis
pegylated granulocyte colony-stimulating factor
Erythroid Precursor Cells
Hematopoiesis
Stem Cell Transplantation
Regulator Genes
Reverse Transcriptase Polymerase Chain Reaction
Granulocytes
Blood Cells
Macrophages
Phenotype

Keywords

  • Autologous stem cell transplantation
  • Pegylated G-CSF
  • Unconjugated G-CSF

ASJC Scopus subject areas

  • Hematology

Cite this

Pegylated granulocyte colony-stimulating factor mobilizes CD34+ cells with different stem and progenitor subsets and distinct functional properties in comparison with unconjugated granulocyte colony-stimulating factor. / Bruns, Ingmar; Steidl, Ulrich G.; Fischer, Johannes C.; Czibere, Akos; Kobbe, Guido; Raschke, Sascha; Singh, Raminder; Fenk, Roland; Rosskopf, Michael; Pechtel, Sabrina; Von Haeseler, Arndt; Wernet, Peter; Tenen, Daniel G.; Haas, Rainer; Kronenwett, Ralf.

In: Haematologica, Vol. 93, No. 3, 03.2008, p. 347-355.

Research output: Contribution to journalArticle

Bruns, I, Steidl, UG, Fischer, JC, Czibere, A, Kobbe, G, Raschke, S, Singh, R, Fenk, R, Rosskopf, M, Pechtel, S, Von Haeseler, A, Wernet, P, Tenen, DG, Haas, R & Kronenwett, R 2008, 'Pegylated granulocyte colony-stimulating factor mobilizes CD34+ cells with different stem and progenitor subsets and distinct functional properties in comparison with unconjugated granulocyte colony-stimulating factor', Haematologica, vol. 93, no. 3, pp. 347-355. https://doi.org/10.3324/haematol.12081
Bruns, Ingmar ; Steidl, Ulrich G. ; Fischer, Johannes C. ; Czibere, Akos ; Kobbe, Guido ; Raschke, Sascha ; Singh, Raminder ; Fenk, Roland ; Rosskopf, Michael ; Pechtel, Sabrina ; Von Haeseler, Arndt ; Wernet, Peter ; Tenen, Daniel G. ; Haas, Rainer ; Kronenwett, Ralf. / Pegylated granulocyte colony-stimulating factor mobilizes CD34+ cells with different stem and progenitor subsets and distinct functional properties in comparison with unconjugated granulocyte colony-stimulating factor. In: Haematologica. 2008 ; Vol. 93, No. 3. pp. 347-355.
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abstract = "Background: Pegylated granulocyte colony-stimulating factor (G-CSF) has recently been introduced as a new compound for mobilization of CD34+ hematopoietic stem and progenitor cells. In this study, we compared the molecular and functional characteristics of CD34+ cells mobilized by pegylated G-CSF with those mobilized by unconjugated G-CSF. Design and Methods: Gene expression of immunomagnetically enriched CD34+ cells from leukapheresis products of patients who were given pegylated-G-CSF or unconjugated G-CSF was analyzed using Affymetrix HG Focus microarrays and quantitative reverse transcriptase polymerase chain reaction. Flow cytometry and fluorescence activated cell sorting was conducted to assess the CD34 + subset composition and to obtain Lin-, CD34+, CD38- hematopoietic stem cells. Cell cycle assays and clonogenic assays were performed for functional corroboration. Results: Pegylated G-CSF and unconjugated G-CSF mobilized CD34+ and hematopoietic stem cells with different molecular phenotypes and functional properties. The CD34+ cells mobilized by pegylated G-CSF had higher expression levels of genes indicative of early hematopoiesis, including HOXA9, MEIS1 and GATA3. We found lower expression of genes characteristic of erythroid and later stages of myeloid differentiation and a lower functional burst-forming unit erythroid/colony-forming unit-granulocyte-macrophage ratio. Consistently, greater numbers of hematopoietic stem cells and common myeloid progenitors and fewer megakaryocyte-erthrocyte progenitors were found in the pegylated-G-CSF-mobilized CD34+ cells. Additionally, sorted pegylated-G-CSF-mobilized hematopoietic stem cells displayed higher expression of HOXA9 in comparison to G-CSF-mobilized hematopoietic stem cells. In line with the gene expression data, CD34+ cells mobilized by pegylated G-CSF, as well as sorted hematopoietic stem cells, showed a significantly greater cell cycle activity. Conclusions: Stimulation with pegylated-G-CSF or G-CSF results in different expression of key regulatory genes and different functional properties of mobilized hematopoietic stem cells as well as their progeny, a finding that might be relevant for the application of these cells in blood stem cell transplantation.",
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AU - Bruns, Ingmar

AU - Steidl, Ulrich G.

AU - Fischer, Johannes C.

AU - Czibere, Akos

AU - Kobbe, Guido

AU - Raschke, Sascha

AU - Singh, Raminder

AU - Fenk, Roland

AU - Rosskopf, Michael

AU - Pechtel, Sabrina

AU - Von Haeseler, Arndt

AU - Wernet, Peter

AU - Tenen, Daniel G.

AU - Haas, Rainer

AU - Kronenwett, Ralf

PY - 2008/3

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N2 - Background: Pegylated granulocyte colony-stimulating factor (G-CSF) has recently been introduced as a new compound for mobilization of CD34+ hematopoietic stem and progenitor cells. In this study, we compared the molecular and functional characteristics of CD34+ cells mobilized by pegylated G-CSF with those mobilized by unconjugated G-CSF. Design and Methods: Gene expression of immunomagnetically enriched CD34+ cells from leukapheresis products of patients who were given pegylated-G-CSF or unconjugated G-CSF was analyzed using Affymetrix HG Focus microarrays and quantitative reverse transcriptase polymerase chain reaction. Flow cytometry and fluorescence activated cell sorting was conducted to assess the CD34 + subset composition and to obtain Lin-, CD34+, CD38- hematopoietic stem cells. Cell cycle assays and clonogenic assays were performed for functional corroboration. Results: Pegylated G-CSF and unconjugated G-CSF mobilized CD34+ and hematopoietic stem cells with different molecular phenotypes and functional properties. The CD34+ cells mobilized by pegylated G-CSF had higher expression levels of genes indicative of early hematopoiesis, including HOXA9, MEIS1 and GATA3. We found lower expression of genes characteristic of erythroid and later stages of myeloid differentiation and a lower functional burst-forming unit erythroid/colony-forming unit-granulocyte-macrophage ratio. Consistently, greater numbers of hematopoietic stem cells and common myeloid progenitors and fewer megakaryocyte-erthrocyte progenitors were found in the pegylated-G-CSF-mobilized CD34+ cells. Additionally, sorted pegylated-G-CSF-mobilized hematopoietic stem cells displayed higher expression of HOXA9 in comparison to G-CSF-mobilized hematopoietic stem cells. In line with the gene expression data, CD34+ cells mobilized by pegylated G-CSF, as well as sorted hematopoietic stem cells, showed a significantly greater cell cycle activity. Conclusions: Stimulation with pegylated-G-CSF or G-CSF results in different expression of key regulatory genes and different functional properties of mobilized hematopoietic stem cells as well as their progeny, a finding that might be relevant for the application of these cells in blood stem cell transplantation.

AB - Background: Pegylated granulocyte colony-stimulating factor (G-CSF) has recently been introduced as a new compound for mobilization of CD34+ hematopoietic stem and progenitor cells. In this study, we compared the molecular and functional characteristics of CD34+ cells mobilized by pegylated G-CSF with those mobilized by unconjugated G-CSF. Design and Methods: Gene expression of immunomagnetically enriched CD34+ cells from leukapheresis products of patients who were given pegylated-G-CSF or unconjugated G-CSF was analyzed using Affymetrix HG Focus microarrays and quantitative reverse transcriptase polymerase chain reaction. Flow cytometry and fluorescence activated cell sorting was conducted to assess the CD34 + subset composition and to obtain Lin-, CD34+, CD38- hematopoietic stem cells. Cell cycle assays and clonogenic assays were performed for functional corroboration. Results: Pegylated G-CSF and unconjugated G-CSF mobilized CD34+ and hematopoietic stem cells with different molecular phenotypes and functional properties. The CD34+ cells mobilized by pegylated G-CSF had higher expression levels of genes indicative of early hematopoiesis, including HOXA9, MEIS1 and GATA3. We found lower expression of genes characteristic of erythroid and later stages of myeloid differentiation and a lower functional burst-forming unit erythroid/colony-forming unit-granulocyte-macrophage ratio. Consistently, greater numbers of hematopoietic stem cells and common myeloid progenitors and fewer megakaryocyte-erthrocyte progenitors were found in the pegylated-G-CSF-mobilized CD34+ cells. Additionally, sorted pegylated-G-CSF-mobilized hematopoietic stem cells displayed higher expression of HOXA9 in comparison to G-CSF-mobilized hematopoietic stem cells. In line with the gene expression data, CD34+ cells mobilized by pegylated G-CSF, as well as sorted hematopoietic stem cells, showed a significantly greater cell cycle activity. Conclusions: Stimulation with pegylated-G-CSF or G-CSF results in different expression of key regulatory genes and different functional properties of mobilized hematopoietic stem cells as well as their progeny, a finding that might be relevant for the application of these cells in blood stem cell transplantation.

KW - Autologous stem cell transplantation

KW - Pegylated G-CSF

KW - Unconjugated G-CSF

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