GATA-4 promotes myocardial transdifferentiation of mesenchymal stromal cells via up-regulating IGFBP-4

Hongxia Li, Shi Zuo, Zeeshan Pasha, Bin Yu, Zhisong He, Yigang Wang, Xiangjun Yang, Muhammad Ashraf, Meifeng Xu

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

Background aims. GATA-4 is a cardiac transcription factor and plays an important role in cell lineage differentiation during development. We investigated whether overexpression of GATA-4 increases adult mesenchymal stromal cell (MSC) transdifferentiation into a cardiac phenotype in vitro. Methods. MSC were harvested from rat bone marrow (BM) and transduced with GATA-4 (MSCGATA-4) using a murine stem cell virus (pMSCV) retroviral expression system. Gene expression in MSCGATA-4 was analyzed using quantitative reverse transcriptionpolymerase chain reaction (RT-PCR) and Western blotting. Native cardiomyocytes (CM) were isolated from ventricles of neonatal rats. Myocardial transdifferentiation of MSC was determined by immunostaining and electrophysiologic recording. The transdifferentiation rate was calculated directly from flow cytometery. Results. The expression of cardiac genes, including brain natriuretic peptide (BNP), Islet-1 and α-sarcomeric actinin (α-SA), was up-regulated in MSCGATA-4 compared with control cells that were transfected with Green Fluorescent Protein (GFP) only (MSCNull). At the same time, insulin-like growth factor-binding protein (IGFBP)-4 was significantly up-regulated in MSCGATA-4. A synchronous beating of MSC with native CM was detected and an action potential was recorded. Some GFP + cells were positive for α-SA staining after MSC were co-cultured with native CM for 7 days. The transdifferentiation rate was significantly higher in MSCGATA-4. Functional studies indicated that the differentiation potential of MSCGATA-4 was decreased by knockdown of IGFBP-4. Conclusions. Overexpression of GATA-4 significantly increases MSC differentiation into a myocardial phenotype, which might be associated with the up-regulation of IGFBP-4.

Original languageEnglish (US)
Pages (from-to)1057-1065
Number of pages9
JournalCytotherapy
Volume13
Issue number9
DOIs
StatePublished - Jan 1 2011
Externally publishedYes

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Insulin-Like Growth Factor Binding Protein 4
Mesenchymal Stromal Cells
Cardiac Myocytes
Green Fluorescent Proteins
Cell Differentiation
Cell Transdifferentiation
Phenotype
Actinin
Gene Expression
Brain Natriuretic Peptide
Cell Lineage
Action Potentials
Transcription Factors
Up-Regulation
Stem Cells
Bone Marrow
Western Blotting
Staining and Labeling
Viruses

Keywords

  • GATA-4
  • cardiomyocyte
  • co-culture
  • genetic engineering
  • insulin-like growth factor-binding protein-4
  • stem cell

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology
  • Oncology
  • Genetics(clinical)
  • Cell Biology
  • Transplantation
  • Cancer Research

Cite this

GATA-4 promotes myocardial transdifferentiation of mesenchymal stromal cells via up-regulating IGFBP-4. / Li, Hongxia; Zuo, Shi; Pasha, Zeeshan; Yu, Bin; He, Zhisong; Wang, Yigang; Yang, Xiangjun; Ashraf, Muhammad; Xu, Meifeng.

In: Cytotherapy, Vol. 13, No. 9, 01.01.2011, p. 1057-1065.

Research output: Contribution to journalArticle

Li, Hongxia ; Zuo, Shi ; Pasha, Zeeshan ; Yu, Bin ; He, Zhisong ; Wang, Yigang ; Yang, Xiangjun ; Ashraf, Muhammad ; Xu, Meifeng. / GATA-4 promotes myocardial transdifferentiation of mesenchymal stromal cells via up-regulating IGFBP-4. In: Cytotherapy. 2011 ; Vol. 13, No. 9. pp. 1057-1065.
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T1 - GATA-4 promotes myocardial transdifferentiation of mesenchymal stromal cells via up-regulating IGFBP-4

AU - Li, Hongxia

AU - Zuo, Shi

AU - Pasha, Zeeshan

AU - Yu, Bin

AU - He, Zhisong

AU - Wang, Yigang

AU - Yang, Xiangjun

AU - Ashraf, Muhammad

AU - Xu, Meifeng

PY - 2011/1/1

Y1 - 2011/1/1

N2 - Background aims. GATA-4 is a cardiac transcription factor and plays an important role in cell lineage differentiation during development. We investigated whether overexpression of GATA-4 increases adult mesenchymal stromal cell (MSC) transdifferentiation into a cardiac phenotype in vitro. Methods. MSC were harvested from rat bone marrow (BM) and transduced with GATA-4 (MSCGATA-4) using a murine stem cell virus (pMSCV) retroviral expression system. Gene expression in MSCGATA-4 was analyzed using quantitative reverse transcriptionpolymerase chain reaction (RT-PCR) and Western blotting. Native cardiomyocytes (CM) were isolated from ventricles of neonatal rats. Myocardial transdifferentiation of MSC was determined by immunostaining and electrophysiologic recording. The transdifferentiation rate was calculated directly from flow cytometery. Results. The expression of cardiac genes, including brain natriuretic peptide (BNP), Islet-1 and α-sarcomeric actinin (α-SA), was up-regulated in MSCGATA-4 compared with control cells that were transfected with Green Fluorescent Protein (GFP) only (MSCNull). At the same time, insulin-like growth factor-binding protein (IGFBP)-4 was significantly up-regulated in MSCGATA-4. A synchronous beating of MSC with native CM was detected and an action potential was recorded. Some GFP + cells were positive for α-SA staining after MSC were co-cultured with native CM for 7 days. The transdifferentiation rate was significantly higher in MSCGATA-4. Functional studies indicated that the differentiation potential of MSCGATA-4 was decreased by knockdown of IGFBP-4. Conclusions. Overexpression of GATA-4 significantly increases MSC differentiation into a myocardial phenotype, which might be associated with the up-regulation of IGFBP-4.

AB - Background aims. GATA-4 is a cardiac transcription factor and plays an important role in cell lineage differentiation during development. We investigated whether overexpression of GATA-4 increases adult mesenchymal stromal cell (MSC) transdifferentiation into a cardiac phenotype in vitro. Methods. MSC were harvested from rat bone marrow (BM) and transduced with GATA-4 (MSCGATA-4) using a murine stem cell virus (pMSCV) retroviral expression system. Gene expression in MSCGATA-4 was analyzed using quantitative reverse transcriptionpolymerase chain reaction (RT-PCR) and Western blotting. Native cardiomyocytes (CM) were isolated from ventricles of neonatal rats. Myocardial transdifferentiation of MSC was determined by immunostaining and electrophysiologic recording. The transdifferentiation rate was calculated directly from flow cytometery. Results. The expression of cardiac genes, including brain natriuretic peptide (BNP), Islet-1 and α-sarcomeric actinin (α-SA), was up-regulated in MSCGATA-4 compared with control cells that were transfected with Green Fluorescent Protein (GFP) only (MSCNull). At the same time, insulin-like growth factor-binding protein (IGFBP)-4 was significantly up-regulated in MSCGATA-4. A synchronous beating of MSC with native CM was detected and an action potential was recorded. Some GFP + cells were positive for α-SA staining after MSC were co-cultured with native CM for 7 days. The transdifferentiation rate was significantly higher in MSCGATA-4. Functional studies indicated that the differentiation potential of MSCGATA-4 was decreased by knockdown of IGFBP-4. Conclusions. Overexpression of GATA-4 significantly increases MSC differentiation into a myocardial phenotype, which might be associated with the up-regulation of IGFBP-4.

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KW - co-culture

KW - genetic engineering

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