A comparison of the pro-angiogenic potential of human induced pluripotent stem cell derived endothelial cells and induced endothelial cells in a murine model of peripheral arterial disease

Zoe E Clayton; Gloria S C Yuen; Sara Sadeghipour; Jack D Hywood; Jack W T Wong; Ngan F Huang; Martin K C Ng; John P Cooke; Sanjay Patel

doi:10.1016/j.ijcard.2017.01.125

A comparison of the pro-angiogenic potential of human induced pluripotent stem cell derived endothelial cells and induced endothelial cells in a murine model of peripheral arterial disease

Int J Cardiol. 2017 May 1:234:81-89. doi: 10.1016/j.ijcard.2017.01.125. Epub 2017 Jan 31.

Authors

Zoe E Clayton¹, Gloria S C Yuen², Sara Sadeghipour³, Jack D Hywood⁴, Jack W T Wong⁵, Ngan F Huang⁶, Martin K C Ng⁷, John P Cooke⁸, Sanjay Patel⁹

Affiliations

¹ Heart Research Institute, 7 Eliza Street, Newtown, NSW 2042, Australia; Sydney Medical School, University of Sydney, Camperdown, NSW 2006, Australia. Electronic address: [email protected].
² Heart Research Institute, 7 Eliza Street, Newtown, NSW 2042, Australia; Sydney Medical School, University of Sydney, Camperdown, NSW 2006, Australia. Electronic address: [email protected].
³ Heart Research Institute, 7 Eliza Street, Newtown, NSW 2042, Australia. Electronic address: [email protected].
⁴ Heart Research Institute, 7 Eliza Street, Newtown, NSW 2042, Australia; Sydney Medical School, University of Sydney, Camperdown, NSW 2006, Australia. Electronic address: [email protected].
⁵ Department of Cardiovascular Sciences, Houston Methodist Research Institute, 6670 Bertner Ave, Houston, TX 77030, USA. Electronic address: [email protected].
⁶ Department of Cardiothoracic Surgery, Stanford University, Stanford, CA. Stanford Cardiovascular Institute, Stanford, CA, United States. Electronic address: [email protected].
⁷ Heart Research Institute, 7 Eliza Street, Newtown, NSW 2042, Australia; Sydney Medical School, University of Sydney, Camperdown, NSW 2006, Australia; Department of Cardiology, Royal Prince Alfred Hospital, Missenden Road, Camperdown, NSW 2050, Australia. Electronic address: [email protected].
⁸ Department of Cardiovascular Sciences, Houston Methodist Research Institute, 6670 Bertner Ave, Houston, TX 77030, USA. Electronic address: [email protected].
⁹ Heart Research Institute, 7 Eliza Street, Newtown, NSW 2042, Australia; Sydney Medical School, University of Sydney, Camperdown, NSW 2006, Australia; Department of Cardiology, Royal Prince Alfred Hospital, Missenden Road, Camperdown, NSW 2050, Australia. Electronic address: [email protected].

PMID: 28209385
DOI: 10.1016/j.ijcard.2017.01.125

Abstract

Background: Endothelial cells derived from human induced pluripotent stem cells (iPSC-ECs) promote angiogenesis, and more recently induced endothelial cells (iECs) have been generated via fibroblast trans-differentiation. These cell types have potential as treatments for peripheral arterial disease (PAD). However, it is unknown whether different reprogramming methods produce cells that are equivalent in terms of their pro-angiogenic capabilities.

Objectives: We aimed to directly compare iPSC-ECs and iECs in an animal model of PAD, in order to identify which cell type, if any, displays superior therapeutic potential.

Methods: IPSC-ECs and iECs were generated from human fibroblasts, and transduced with a reporter construct encoding GFP and firefly luciferase for bioluminescence imaging (BLI). Endothelial phenotype was confirmed using in vitro assays. NOD-SCID mice underwent hindlimb ischaemia surgery and received an intramuscular injection of either 1×10⁶ iPSC-ECs, 1×10⁶ iECs or control vehicle only. Perfusion recovery was measured by laser Doppler. Hindlimb muscle samples were taken for histological analyses.

Results: Perfusion recovery was enhanced in iPSC-EC treated mice on day 14 (Control vs. iPSC-EC; 0.35±0.04 vs. 0.54±0.08, p<0.05) and in iEC treated mice on days 7 (Control vs. iEC; 0.23±0.02 vs. 0.44±0.06, p<0.05), 10 (0.31±0.04 vs. 0.64±0.07, p<0.001) and 14 (0.35±0.04 vs. 0.68±0.07, p<0.001) post-treatment. IEC-treated mice also had greater capillary density in the ischaemic gastrocnemius muscle (Control vs. iEC; 125±10 vs. 179±11 capillaries/image; p<0.05). BLI detected iPSC-EC and iEC presence in vivo for two weeks post-treatment.

Conclusions: IPSC-ECs and iECs exhibit similar, but not identical, endothelial functionality and both cell types enhance perfusion recovery after hindlimb ischaemia.

Keywords: Angiogenesis; Cell therapy; Differentiation; Endothelial cells; Peripheral arterial disease; Reprogramming.

MeSH terms

Animals
Cell Differentiation / physiology*
Cells, Cultured
Cellular Reprogramming / physiology
Disease Models, Animal
Endothelial Cells / physiology*
Fibroblasts / physiology
Hindlimb / blood supply
Humans
Induced Pluripotent Stem Cells / physiology
Ischemia* / metabolism
Ischemia* / therapy
Mice
Mice, Inbred NOD
Mice, SCID
Myocardial Perfusion Imaging / methods
Peripheral Arterial Disease* / metabolism
Peripheral Arterial Disease* / physiopathology
Peripheral Arterial Disease* / therapy
Stem Cell Transplantation / methods*
Treatment Outcome