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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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Schematic representation of the overall scheme: generation of recombinant baculovirus (BacAng1), preparation of hybridized baculovirus with TAT/DNA nanoparticles, in vitro human adipose tissue-derived stem cell transduction, and in vivo investigation using direct intramyocardial transplantation of ASC-Ang1 using a rat model with acute myocardial infarction. Transmission electronic microscopic pictures of baculovirus, nanoparticle, and baculovirus-nanoparticle complex shown in subsets confirm the formation of the nanocomplex (in white arrows).Abbreviations: ASC-Ang1, angiopoietin-1-expressing adipose tissue-derived stem cells; BacAng1, angiopoietin-1-carrying baculovirus; Bac-NPAng1, angiopoietin-1-carrying baculovirus-nanoparticle complex; DNA, deoxyribonucleic acid; ELISA, enzyme-linked immunosorbent assay; NPAng1, angiopoietin-1-carrying nanoparticles; TAT, transactivating transcriptional activator.
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f1-ijn-7-663: Schematic representation of the overall scheme: generation of recombinant baculovirus (BacAng1), preparation of hybridized baculovirus with TAT/DNA nanoparticles, in vitro human adipose tissue-derived stem cell transduction, and in vivo investigation using direct intramyocardial transplantation of ASC-Ang1 using a rat model with acute myocardial infarction. Transmission electronic microscopic pictures of baculovirus, nanoparticle, and baculovirus-nanoparticle complex shown in subsets confirm the formation of the nanocomplex (in white arrows).Abbreviations: ASC-Ang1, angiopoietin-1-expressing adipose tissue-derived stem cells; BacAng1, angiopoietin-1-carrying baculovirus; Bac-NPAng1, angiopoietin-1-carrying baculovirus-nanoparticle complex; DNA, deoxyribonucleic acid; ELISA, enzyme-linked immunosorbent assay; NPAng1, angiopoietin-1-carrying nanoparticles; TAT, transactivating transcriptional activator.

Mentions: It has been reported that VEGF in cooperation with another angiogenic growth factor, Ang1, promotes significant neovascularization, and their combined action leads to a formation of mature and functional vasculature.19–21 Thus, it was hypothesized that the secretion of Ang1 along with naturally releasing VEGF from the genetically modified ASCs, together with their inherent transdifferentiation abilities to cardiomyocytes, can induce a superior synergistic therapeutic effect for myocardial regeneration therapy. Additionally, acute myocardial infarction also induces a high circulating endogenous serum VEGF state.22 With these rationales, Ang1 complementary DNA-carrying recombinant Bac and TAT NPs were generated, a self-assembled binary nanocomplex was prepared by hybridizing the two, and its efficiency to express functionally active Ang1 was determined using optimized transduction protocol with hASCs. The in vivo efficacy of the formulated nanobiohybrid (Bac-NP) system for combined stem cell-gene therapy applications was also evaluated using human Ang1 (hAng1)-expressing genetically modified hASCs for myocardial therapy using an immunocompetent infarcted rat heart model. A schematic presentation of the entire procedure is demonstrated in Figure 1.


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)

Schematic representation of the overall scheme: generation of recombinant baculovirus (BacAng1), preparation of hybridized baculovirus with TAT/DNA nanoparticles, in vitro human adipose tissue-derived stem cell transduction, and in vivo investigation using direct intramyocardial transplantation of ASC-Ang1 using a rat model with acute myocardial infarction. Transmission electronic microscopic pictures of baculovirus, nanoparticle, and baculovirus-nanoparticle complex shown in subsets confirm the formation of the nanocomplex (in white arrows).Abbreviations: ASC-Ang1, angiopoietin-1-expressing adipose tissue-derived stem cells; BacAng1, angiopoietin-1-carrying baculovirus; Bac-NPAng1, angiopoietin-1-carrying baculovirus-nanoparticle complex; DNA, deoxyribonucleic acid; ELISA, enzyme-linked immunosorbent assay; NPAng1, angiopoietin-1-carrying nanoparticles; TAT, transactivating transcriptional activator.
© Copyright Policy
Related In: Results  -  Collection

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

f1-ijn-7-663: Schematic representation of the overall scheme: generation of recombinant baculovirus (BacAng1), preparation of hybridized baculovirus with TAT/DNA nanoparticles, in vitro human adipose tissue-derived stem cell transduction, and in vivo investigation using direct intramyocardial transplantation of ASC-Ang1 using a rat model with acute myocardial infarction. Transmission electronic microscopic pictures of baculovirus, nanoparticle, and baculovirus-nanoparticle complex shown in subsets confirm the formation of the nanocomplex (in white arrows).Abbreviations: ASC-Ang1, angiopoietin-1-expressing adipose tissue-derived stem cells; BacAng1, angiopoietin-1-carrying baculovirus; Bac-NPAng1, angiopoietin-1-carrying baculovirus-nanoparticle complex; DNA, deoxyribonucleic acid; ELISA, enzyme-linked immunosorbent assay; NPAng1, angiopoietin-1-carrying nanoparticles; TAT, transactivating transcriptional activator.
Mentions: It has been reported that VEGF in cooperation with another angiogenic growth factor, Ang1, promotes significant neovascularization, and their combined action leads to a formation of mature and functional vasculature.19–21 Thus, it was hypothesized that the secretion of Ang1 along with naturally releasing VEGF from the genetically modified ASCs, together with their inherent transdifferentiation abilities to cardiomyocytes, can induce a superior synergistic therapeutic effect for myocardial regeneration therapy. Additionally, acute myocardial infarction also induces a high circulating endogenous serum VEGF state.22 With these rationales, Ang1 complementary DNA-carrying recombinant Bac and TAT NPs were generated, a self-assembled binary nanocomplex was prepared by hybridizing the two, and its efficiency to express functionally active Ang1 was determined using optimized transduction protocol with hASCs. The in vivo efficacy of the formulated nanobiohybrid (Bac-NP) system for combined stem cell-gene therapy applications was also evaluated using human Ang1 (hAng1)-expressing genetically modified hASCs for myocardial therapy using an immunocompetent infarcted rat heart model. A schematic presentation of the entire procedure is demonstrated in Figure 1.

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