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Angiopoietin-1-expressing adipose stem cells genetically modified with baculovirus nanocomplex: investigation in rat heart with acute infarction.

Paul A, Nayan M, Khan AA, Shum-Tim D, Prakash S - Int J Nanomedicine (2012)

Bottom Line: The released hAng1 from hASC-Ang1 demonstrated profound mitotic and anti-apoptotic activities on endothelial cells and cardiomyocytes.The transplanted hASC-Ang1 group showed higher cell retention compared to hASC and control groups.A significant increase in capillary density and reduction in infarct sizes were noted in the infarcted hearts with hASC-Ang1 treatment compared to infarcted hearts treated with hASC or the untreated group.

View Article: PubMed Central - PubMed

Affiliation: Biomedical Technology and Cell Therapy Research Laboratory, Department of Biomedical Engineering, Faculty of Medicine, McGill University, Montreal, QC, Canada.

ABSTRACT
The objective of this study was to develop angiopoietin-1 (Ang1)-expressing genetically modified human adipose tissue derived stem cells (hASCs) for myocardial therapy. For this, an efficient gene delivery system using recombinant baculovirus complexed with cell penetrating transactivating transcriptional activator TAT peptide/deoxyribonucleic acid nanoparticles (Bac-NP), through ionic interactions, was used. It was hypothesized that the hybrid Bac- NP(Ang1) system can efficiently transduce hASCs and induces favorable therapeutic effects when transplanted in vivo. To evaluate this hypothesis, a rat model with acute myocardial infarction and intramyocardially transplanted Ang1-expressing hASCs (hASC-Ang1), genetically modified by Bac-NP(Ang1), was used. Ang1 is a crucial pro-angiogenic factor for vascular maturation and neovasculogenesis. The released hAng1 from hASC-Ang1 demonstrated profound mitotic and anti-apoptotic activities on endothelial cells and cardiomyocytes. The transplanted hASC-Ang1 group showed higher cell retention compared to hASC and control groups. A significant increase in capillary density and reduction in infarct sizes were noted in the infarcted hearts with hASC-Ang1 treatment compared to infarcted hearts treated with hASC or the untreated group. Furthermore, the hASC-Ang1 group showed significantly higher cardiac performance in echocardiography (ejection fraction 46.28% ± 6.3%, P < 0.001 versus control, n = 8) than the hASC group (36.35% ± 5.7%, P < 0.01, n = 8), 28 days post-infarction. The study identified Bac-NP complex as an advanced gene delivery vehicle for stem cells and demonstrated its potential to treat ischemic heart disease with high therapeutic index for combined stem cell-gene therapy strategy.

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(A) Detection of transgene expression (LacZ) in baculovirus transduced hASCs transplanted in the myocardium. (i) Three million cells were transduced with Bac-NPLacZ (multiplicity of infection: 200) and intramyocardially injected in the left ventricular myocardium. (ii) Three days post-transplantation, hearts (n = 3) were harvested and traced for LacZ expression. (iii) Histological analysis of ventricular portion (200×: stained with eosin) shows the transplanted cells expressing the LacZ. Arrows show the transplanted stained hASC-LacZ cells in the periinfarct ventricle region. (B) Higher retention of hASC-Ang1 compared to only hASCs in the left ventricular myocardium 28 days post-transplantation. (i) Polymerase chain reaction products (250 base pairs) specific for the human Y chromosome (DYS390 sequence) as detected in 2% agarose gel. There were clear distinct bands in group hASC-Ang1 (Lane 1 and 1′) and group hASC (Lane 2 and 2′) in all female rat hearts both on day three and day 28, while these bands were absent in no hASC control group (Lane 3 and 3′). Note that these band intensities were much lower in day 28 compared to day three, with group hASC having the least intensity. Representative polymerase chain reaction products from each group are shown here. (ii) ImageJ 1.41 software (National Institutes of Health, Bethesda, MA) analysis of the band intensities show the hASC group has a mean of 0.32 times (n = 3) and group hASC-Ang1 has a mean of 0.55 times (n = 3) band intensities, taking the band intensity of group hASC on day three as 1.0.Notes: Two-way analysis of variance: F = 34.86, P = 0.0004 (treatment groups); F = 974.0, P < 0.0001 (day); F = 49.41, P = 0.0001 (interaction); ***P < 0.001 versus day-matched control hASC.Abbreviations: Bac-NPLacZ, LacZ-carrying baculovirus-nanoparticle complex; hASC, human adipose tissue-derived cell; hASC-Ang1, angiopoietin-1-expressing human adipose tissue-derived cell; hASC-LacZ, LacZ-expressing human adipose tissue-derived cell; NTC, no template control for the polymerase chain reaction experiment.
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f5-ijn-7-663: (A) Detection of transgene expression (LacZ) in baculovirus transduced hASCs transplanted in the myocardium. (i) Three million cells were transduced with Bac-NPLacZ (multiplicity of infection: 200) and intramyocardially injected in the left ventricular myocardium. (ii) Three days post-transplantation, hearts (n = 3) were harvested and traced for LacZ expression. (iii) Histological analysis of ventricular portion (200×: stained with eosin) shows the transplanted cells expressing the LacZ. Arrows show the transplanted stained hASC-LacZ cells in the periinfarct ventricle region. (B) Higher retention of hASC-Ang1 compared to only hASCs in the left ventricular myocardium 28 days post-transplantation. (i) Polymerase chain reaction products (250 base pairs) specific for the human Y chromosome (DYS390 sequence) as detected in 2% agarose gel. There were clear distinct bands in group hASC-Ang1 (Lane 1 and 1′) and group hASC (Lane 2 and 2′) in all female rat hearts both on day three and day 28, while these bands were absent in no hASC control group (Lane 3 and 3′). Note that these band intensities were much lower in day 28 compared to day three, with group hASC having the least intensity. Representative polymerase chain reaction products from each group are shown here. (ii) ImageJ 1.41 software (National Institutes of Health, Bethesda, MA) analysis of the band intensities show the hASC group has a mean of 0.32 times (n = 3) and group hASC-Ang1 has a mean of 0.55 times (n = 3) band intensities, taking the band intensity of group hASC on day three as 1.0.Notes: Two-way analysis of variance: F = 34.86, P = 0.0004 (treatment groups); F = 974.0, P < 0.0001 (day); F = 49.41, P = 0.0001 (interaction); ***P < 0.001 versus day-matched control hASC.Abbreviations: Bac-NPLacZ, LacZ-carrying baculovirus-nanoparticle complex; hASC, human adipose tissue-derived cell; hASC-Ang1, angiopoietin-1-expressing human adipose tissue-derived cell; hASC-LacZ, LacZ-expressing human adipose tissue-derived cell; NTC, no template control for the polymerase chain reaction experiment.

Mentions: To analyze whether the xenotransplanted genetically modified hASCs were able to survive and express the transgene in the rat heart, hearts (n = 3) were myocardially infarcted and injected with hASCs transduced with Bac-NPLacZ in the left ventricular region, as described earlier. Three days post-transplantation, the rat hearts were harvested and ventricular tissue samples were stained with X-gal in order to trace the transplanted LacZ-expressing hASCs in the peri-infarct region of the heart. Figure 5A confirms that the transplanted cells were able to survive and express the transgene in the heart.


Angiopoietin-1-expressing adipose stem cells genetically modified with baculovirus nanocomplex: investigation in rat heart with acute infarction.

Paul A, Nayan M, Khan AA, Shum-Tim D, Prakash S - Int J Nanomedicine (2012)

(A) Detection of transgene expression (LacZ) in baculovirus transduced hASCs transplanted in the myocardium. (i) Three million cells were transduced with Bac-NPLacZ (multiplicity of infection: 200) and intramyocardially injected in the left ventricular myocardium. (ii) Three days post-transplantation, hearts (n = 3) were harvested and traced for LacZ expression. (iii) Histological analysis of ventricular portion (200×: stained with eosin) shows the transplanted cells expressing the LacZ. Arrows show the transplanted stained hASC-LacZ cells in the periinfarct ventricle region. (B) Higher retention of hASC-Ang1 compared to only hASCs in the left ventricular myocardium 28 days post-transplantation. (i) Polymerase chain reaction products (250 base pairs) specific for the human Y chromosome (DYS390 sequence) as detected in 2% agarose gel. There were clear distinct bands in group hASC-Ang1 (Lane 1 and 1′) and group hASC (Lane 2 and 2′) in all female rat hearts both on day three and day 28, while these bands were absent in no hASC control group (Lane 3 and 3′). Note that these band intensities were much lower in day 28 compared to day three, with group hASC having the least intensity. Representative polymerase chain reaction products from each group are shown here. (ii) ImageJ 1.41 software (National Institutes of Health, Bethesda, MA) analysis of the band intensities show the hASC group has a mean of 0.32 times (n = 3) and group hASC-Ang1 has a mean of 0.55 times (n = 3) band intensities, taking the band intensity of group hASC on day three as 1.0.Notes: Two-way analysis of variance: F = 34.86, P = 0.0004 (treatment groups); F = 974.0, P < 0.0001 (day); F = 49.41, P = 0.0001 (interaction); ***P < 0.001 versus day-matched control hASC.Abbreviations: Bac-NPLacZ, LacZ-carrying baculovirus-nanoparticle complex; hASC, human adipose tissue-derived cell; hASC-Ang1, angiopoietin-1-expressing human adipose tissue-derived cell; hASC-LacZ, LacZ-expressing human adipose tissue-derived cell; NTC, no template control for the polymerase chain reaction experiment.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3278230&req=5

f5-ijn-7-663: (A) Detection of transgene expression (LacZ) in baculovirus transduced hASCs transplanted in the myocardium. (i) Three million cells were transduced with Bac-NPLacZ (multiplicity of infection: 200) and intramyocardially injected in the left ventricular myocardium. (ii) Three days post-transplantation, hearts (n = 3) were harvested and traced for LacZ expression. (iii) Histological analysis of ventricular portion (200×: stained with eosin) shows the transplanted cells expressing the LacZ. Arrows show the transplanted stained hASC-LacZ cells in the periinfarct ventricle region. (B) Higher retention of hASC-Ang1 compared to only hASCs in the left ventricular myocardium 28 days post-transplantation. (i) Polymerase chain reaction products (250 base pairs) specific for the human Y chromosome (DYS390 sequence) as detected in 2% agarose gel. There were clear distinct bands in group hASC-Ang1 (Lane 1 and 1′) and group hASC (Lane 2 and 2′) in all female rat hearts both on day three and day 28, while these bands were absent in no hASC control group (Lane 3 and 3′). Note that these band intensities were much lower in day 28 compared to day three, with group hASC having the least intensity. Representative polymerase chain reaction products from each group are shown here. (ii) ImageJ 1.41 software (National Institutes of Health, Bethesda, MA) analysis of the band intensities show the hASC group has a mean of 0.32 times (n = 3) and group hASC-Ang1 has a mean of 0.55 times (n = 3) band intensities, taking the band intensity of group hASC on day three as 1.0.Notes: Two-way analysis of variance: F = 34.86, P = 0.0004 (treatment groups); F = 974.0, P < 0.0001 (day); F = 49.41, P = 0.0001 (interaction); ***P < 0.001 versus day-matched control hASC.Abbreviations: Bac-NPLacZ, LacZ-carrying baculovirus-nanoparticle complex; hASC, human adipose tissue-derived cell; hASC-Ang1, angiopoietin-1-expressing human adipose tissue-derived cell; hASC-LacZ, LacZ-expressing human adipose tissue-derived cell; NTC, no template control for the polymerase chain reaction experiment.
Mentions: To analyze whether the xenotransplanted genetically modified hASCs were able to survive and express the transgene in the rat heart, hearts (n = 3) were myocardially infarcted and injected with hASCs transduced with Bac-NPLacZ in the left ventricular region, as described earlier. Three days post-transplantation, the rat hearts were harvested and ventricular tissue samples were stained with X-gal in order to trace the transplanted LacZ-expressing hASCs in the peri-infarct region of the heart. Figure 5A confirms that the transplanted cells were able to survive and express the transgene in the heart.

Bottom Line: The released hAng1 from hASC-Ang1 demonstrated profound mitotic and anti-apoptotic activities on endothelial cells and cardiomyocytes.The transplanted hASC-Ang1 group showed higher cell retention compared to hASC and control groups.A significant increase in capillary density and reduction in infarct sizes were noted in the infarcted hearts with hASC-Ang1 treatment compared to infarcted hearts treated with hASC or the untreated group.

View Article: PubMed Central - PubMed

Affiliation: Biomedical Technology and Cell Therapy Research Laboratory, Department of Biomedical Engineering, Faculty of Medicine, McGill University, Montreal, QC, Canada.

ABSTRACT
The objective of this study was to develop angiopoietin-1 (Ang1)-expressing genetically modified human adipose tissue derived stem cells (hASCs) for myocardial therapy. For this, an efficient gene delivery system using recombinant baculovirus complexed with cell penetrating transactivating transcriptional activator TAT peptide/deoxyribonucleic acid nanoparticles (Bac-NP), through ionic interactions, was used. It was hypothesized that the hybrid Bac- NP(Ang1) system can efficiently transduce hASCs and induces favorable therapeutic effects when transplanted in vivo. To evaluate this hypothesis, a rat model with acute myocardial infarction and intramyocardially transplanted Ang1-expressing hASCs (hASC-Ang1), genetically modified by Bac-NP(Ang1), was used. Ang1 is a crucial pro-angiogenic factor for vascular maturation and neovasculogenesis. The released hAng1 from hASC-Ang1 demonstrated profound mitotic and anti-apoptotic activities on endothelial cells and cardiomyocytes. The transplanted hASC-Ang1 group showed higher cell retention compared to hASC and control groups. A significant increase in capillary density and reduction in infarct sizes were noted in the infarcted hearts with hASC-Ang1 treatment compared to infarcted hearts treated with hASC or the untreated group. Furthermore, the hASC-Ang1 group showed significantly higher cardiac performance in echocardiography (ejection fraction 46.28% ± 6.3%, P < 0.001 versus control, n = 8) than the hASC group (36.35% ± 5.7%, P < 0.01, n = 8), 28 days post-infarction. The study identified Bac-NP complex as an advanced gene delivery vehicle for stem cells and demonstrated its potential to treat ischemic heart disease with high therapeutic index for combined stem cell-gene therapy strategy.

Show MeSH
Related in: MedlinePlus