Transient disruption of autocrine TGF-beta signaling leads to enhanced survival and proliferation potential in single primitive human hemopoietic progenitor cells

• Published in: The Journal of immunology (1950) Vol. 168; no. 2; pp. 755 - 762
• Main Authors: Fan, Xiaolong, Valdimarsdottir, Gudrun, Larsson, Jonas, Brun, Ann, Magnusson, Mattias, Jacobsen, Sten Eirik, ten Dijke, Peter, Karlsson, Stefan
• Format: Journal Article
• Language: English
• Published: United States 15.01.2002
• Subjects: Adenoviridae - genetics; ADP-ribosyl Cyclase; ADP-ribosyl Cyclase 1; Animal; Animals; Antigens CD34 : biosynthesis; Antigens Differentiation : biosynthesis; Antigens, CD; Autocrine Communication - genetics; Basic Medicine; Cattle; CD34 antigen; CD38 antigen; Cell Cycle - genetics; Cell Division - genetics; Cell Lineage - genetics; Cell Survival - genetics; Cells Cultured; Culture Media Serum-Free; Genetic Vectors - biosynthesis; HeLa Cells; Hematopoietic Stem Cells - cytology; Hematopoietic Stem Cells - immunology; Hematopoietic Stem Cells - metabolism; Hematopoietic Stem Cells : cytology : immunology : metabolism; Human; Humans; Immunologi inom det medicinska området; Immunology in the medical area; Immunophenotyping; K562 Cells; Lin antigen; Medical and Health Sciences; Medicin och hälsovetenskap; Medicinska och farmaceutiska grundvetenskaper; Membrane Glycoproteins; NAD+ Nucleosidase - biosynthesis; Signal Transduction - genetics; Signal Transduction - physiology; Signal Transduction : genetics : physiology; Support Non-U.S. Gov't; Transduction Genetic; Transforming Growth Factor beta - antagonists & inhibitors; Transforming Growth Factor beta - genetics; Transforming Growth Factor beta - pharmacology; Transforming Growth Factor beta - physiology; Transforming Growth Factor beta : antagonists & inhibitors : genetics : pharmacology : physiology
• ISSN: 0022-1767; 1550-6606; 1550-6606
• DOI: 10.4049/jimmunol.168.2.755

Hemopoietic stem cells (HSCs) are maintained at relative quiescence by the balance between the positive and negative regulatory factors that stimulate or inhibit their proliferation. Blocking the action of negative regulatory factors may provide a new approach for inducing HSCs into proliferation. A variety of studies have suggested that TGF-beta negatively regulates cell cycle progression of HSCs. In this study, a dominant negatively acting mutant of TGF-beta type II receptor (TbetaRIIDN) was transiently expressed in HSCs by using adenoviral vector-mediated gene delivery, such that the effects of disrupting the autocrine TGF-beta signaling in HSCs can be directly examined at a single cell level. Adenoviral vectors allowing the expression of TbetaRIIDN and green fluorescence protein in the same CD34(+)CD38(-)Lin(-) cells were constructed. Overexpression of TbetaRIIDN specifically disrupted TGF-beta-mediated signaling. Autocrine TGF-beta signaling in CD34(+)CD38(-)Lin(-) cells was studied in single cell assays under serum-free conditions. Transient blockage of autocrine TGF-beta signaling in CD34(+)CD38(-)Lin(-) cells enhanced their survival. Furthermore, the overall proliferation potential and proliferation kinetics in these cells were significantly enhanced compared with the CD34(+)CD38(-)Lin(-) cells expressing green fluorescence protein alone. Therefore, we have successfully blocked the autocrine TGF-beta-negative regulatory loop of primitive hemopoietic progenitor cells.