Reversal of ischemic cardiomyopathy with Sca-1+ stem cells modified with multiple growth factors.

<h4>Background</h4>We hypothesized that bone marrow derived Sca-1+ stem cells (BM Sca-1+) transduced with multiple therapeutic cytokines with diverse effects will induce faster angiomyogenic differentiation in the infarcted myocardium.<h4>Methods and results</h4>BM Sca-1+ wer...

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Autores principales:	Ning Li, Zeeshan Pasha, Muhammad Ashraf
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2014
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/6643509ca2cd41c89e9a1f6b6c753664
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spelling	oai:doaj.org-article:6643509ca2cd41c89e9a1f6b6c7536642021-11-18T08:24:55ZReversal of ischemic cardiomyopathy with Sca-1+ stem cells modified with multiple growth factors.1932-620310.1371/journal.pone.0093645https://doaj.org/article/6643509ca2cd41c89e9a1f6b6c7536642014-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/24705272/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203<h4>Background</h4>We hypothesized that bone marrow derived Sca-1+ stem cells (BM Sca-1+) transduced with multiple therapeutic cytokines with diverse effects will induce faster angiomyogenic differentiation in the infarcted myocardium.<h4>Methods and results</h4>BM Sca-1+ were purified from transgenic male mice expressing GFP. Plasmids encoding for select quartet of growth factors, i.e., human IGF-1, VEGF, SDF-1α and HGF were prepared and used for genetic modification of Sca-1+ cells (GFSca-1+). Scramble transfected cells (ScSca-1+) were used as a control. RT-PCR and western blotting showed significantly higher expression of the growth factors in GFSca-1+. Besides the quartet of the therapeutic growth factors, PCR based growth factor array showed upregulation of multiple angiogenic and prosurvival factors such as Ang-1, Ang-2, MMP9, Cx43, BMP2, BMP5, FGF2, and NGF in GFSca-1+ (p<0.01 vs ScSca-1+). LDH and TUNEL assays showed enhanced survival of GFSca-1+ under lethal anoxia (p<0.01 vs ScSca-1+). MTS assay showed significant increased cell proliferation in GFSca-1+ (p<0.05 vs ScSca-1+). For in vivo study, female mice were grouped to receive the intramyocardial injection of 15 μl DMEM without cells (group-1) or containing 2.5 × 10(5) ScSca-1+ (group-2) or GFSca-1+ (group-3) immediately after coronary artery ligation. As indicated by Sry gene, a higher survival of GFSca-1+ in group-3 on day 4 (2.3 fold higher vs group-2) was observed with massive mobilization of stem and progenitor cells (cKit+, Mdr1+, Cxcr4+ cells). Heart tissue sections immunostained for actinin and Cx43 at 4 weeks post engraftment showed extensive myofiber formation and expression of gap junctions. Immunostaining for vWF showed increased blood vessel density in both peri-infarct and infarct regions in group-3. Infarct size was attenuated and the global heart function was improved in group-3 as compared to group-2.<h4>Conclusions</h4>Administration of BM Sca-1+ transduced with multiple genes is a novel approach to treat infarcted heart for its regeneration.Ning LiZeeshan PashaMuhammad AshrafPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 9, Iss 4, p e93645 (2014)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Ning Li Zeeshan Pasha Muhammad Ashraf Reversal of ischemic cardiomyopathy with Sca-1+ stem cells modified with multiple growth factors.
description	<h4>Background</h4>We hypothesized that bone marrow derived Sca-1+ stem cells (BM Sca-1+) transduced with multiple therapeutic cytokines with diverse effects will induce faster angiomyogenic differentiation in the infarcted myocardium.<h4>Methods and results</h4>BM Sca-1+ were purified from transgenic male mice expressing GFP. Plasmids encoding for select quartet of growth factors, i.e., human IGF-1, VEGF, SDF-1α and HGF were prepared and used for genetic modification of Sca-1+ cells (GFSca-1+). Scramble transfected cells (ScSca-1+) were used as a control. RT-PCR and western blotting showed significantly higher expression of the growth factors in GFSca-1+. Besides the quartet of the therapeutic growth factors, PCR based growth factor array showed upregulation of multiple angiogenic and prosurvival factors such as Ang-1, Ang-2, MMP9, Cx43, BMP2, BMP5, FGF2, and NGF in GFSca-1+ (p<0.01 vs ScSca-1+). LDH and TUNEL assays showed enhanced survival of GFSca-1+ under lethal anoxia (p<0.01 vs ScSca-1+). MTS assay showed significant increased cell proliferation in GFSca-1+ (p<0.05 vs ScSca-1+). For in vivo study, female mice were grouped to receive the intramyocardial injection of 15 μl DMEM without cells (group-1) or containing 2.5 × 10(5) ScSca-1+ (group-2) or GFSca-1+ (group-3) immediately after coronary artery ligation. As indicated by Sry gene, a higher survival of GFSca-1+ in group-3 on day 4 (2.3 fold higher vs group-2) was observed with massive mobilization of stem and progenitor cells (cKit+, Mdr1+, Cxcr4+ cells). Heart tissue sections immunostained for actinin and Cx43 at 4 weeks post engraftment showed extensive myofiber formation and expression of gap junctions. Immunostaining for vWF showed increased blood vessel density in both peri-infarct and infarct regions in group-3. Infarct size was attenuated and the global heart function was improved in group-3 as compared to group-2.<h4>Conclusions</h4>Administration of BM Sca-1+ transduced with multiple genes is a novel approach to treat infarcted heart for its regeneration.
format	article
author	Ning Li Zeeshan Pasha Muhammad Ashraf
author_facet	Ning Li Zeeshan Pasha Muhammad Ashraf
author_sort	Ning Li
title	Reversal of ischemic cardiomyopathy with Sca-1+ stem cells modified with multiple growth factors.
title_short	Reversal of ischemic cardiomyopathy with Sca-1+ stem cells modified with multiple growth factors.
title_full	Reversal of ischemic cardiomyopathy with Sca-1+ stem cells modified with multiple growth factors.
title_fullStr	Reversal of ischemic cardiomyopathy with Sca-1+ stem cells modified with multiple growth factors.
title_full_unstemmed	Reversal of ischemic cardiomyopathy with Sca-1+ stem cells modified with multiple growth factors.
title_sort	reversal of ischemic cardiomyopathy with sca-1+ stem cells modified with multiple growth factors.
publisher	Public Library of Science (PLoS)
publishDate	2014
url	https://doaj.org/article/6643509ca2cd41c89e9a1f6b6c753664
work_keys_str_mv	AT ningli reversalofischemiccardiomyopathywithsca1stemcellsmodifiedwithmultiplegrowthfactors AT zeeshanpasha reversalofischemiccardiomyopathywithsca1stemcellsmodifiedwithmultiplegrowthfactors AT muhammadashraf reversalofischemiccardiomyopathywithsca1stemcellsmodifiedwithmultiplegrowthfactors
_version_	1718421805833650176

Reversal of ischemic cardiomyopathy with Sca-1+ stem cells modified with multiple growth factors.

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