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Non-hypoxic stabilization of HIF-Iα during coordinated interaction between Akt and angiopoietin-1 enhances endothelial commitment of bone marrow stem cells

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Abstract

We previously reported that mesenchymal stem cells (MSC) co-expressing Akt and angiopoietin-1 (Ang-1) preserved infarcted heart function via angiomyogenesis. The present study determined the mechanism of co-overexpression of Akt and Ang-1 in promoting endothelial commitment of MSC. The cells were transduced with vectors encoding for Akt (AktMSC), Ang-1 (Ang-1MSC), and both Akt and Ang-1 (AAMSC) using Empty vector transduced MSC (EmpMSC) as control. Molecular studies indicated a coordinated interaction between Akt and Ang-1 in AAMSC and led to non-hypoxic stabilization of hypoxia inducible factor-1α (HIF-Iα) which accentuated under 4-h anoxia. We also observed HIF-Iα dependent induction of hemeoxygenase-1, endothelial specific markers and VEGF in AAMSC. Vascular commitment of AAMSC was confirmed by immunostaining, Western blotting and flow cytometry for endothelial specific early and late markers including Flt1, Flk1, Tie2, VCAM-1, and von Willebrand Factor-VIII (vWF-VIII) in HIF-Iα dependent fashion besides exhibiting higher emigrational activity and angiogenesis in vitro. AAMSC transplanted into rat model of myocardial infarction showed higher Flk1 and Flt1 positivity and also promoted intrinsic Flk1+ and Flt1+ cell mobilization into the infarcted heart. Given the ease of availability of MSC and simplicity of approach to co-overexpress Ang-1 and Akt to enhance their endothelial commitment, the strategy will be significant for cellular angiogenesis to treat ischemic heart.

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Abbreviations

Ang-1:

Angiopoietin-1

BM:

Bone marrow

DMEM:

Dulbecco’s Modified Eagle Medium

HIF-Iα:

Hypoxia inducible factor-1α

HO-1:

Hemeoxygenase-1

AAMSC:

Mesenchymal stem cells overexpressing Akt and angiopoietin-1

EmpMSC:

Empty vector trnasduced MSC

MSC:

Mesenchymal stem cells

PGF:

Placental growth factor

VEGF:

Vascular endothelial growth factor

Sc:

Scramble

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Acknowledgments

This work was supported by NIH Grants# [R37 HL074272;HL-080686;HL-087246 (M.A) and HL-087288;HL-089535; HL106190-01 (Kh.H.H)].

Conflict of Interest Disclosure Statement

Nothing to disclose.

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Authors and Affiliations

  1. Department of Pathology, University of Cincinnati Medical Center, 231-Albert Sabin Way, Cincinnati, OH, 45267-0529, USA

    Vien Khach Lai, Muhammad Rizwan Afzal, Muhammad Ashraf, Shujia Jiang & Husnain Kh Haider

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  1. Vien Khach Lai
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  2. Muhammad Rizwan Afzal
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  3. Muhammad Ashraf
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  4. Shujia Jiang
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  5. Husnain Kh Haider
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Correspondence to Husnain Kh Haider.

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Lai, V.K., Afzal, M.R., Ashraf, M. et al. Non-hypoxic stabilization of HIF-Iα during coordinated interaction between Akt and angiopoietin-1 enhances endothelial commitment of bone marrow stem cells. J Mol Med 90, 719–730 (2012). https://doi.org/10.1007/s00109-011-0852-1

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