Pub Date : 2003-03-01DOI: 10.1163/156855803762295413
J. Smith, Suzanne Thomas, R. Coffin, D. Latchman
In this paper, we have studied the potential interactions of replication incompetent herpes virus vectors with replication-competent virus both in vitro and in vivo. This might be thought to be particularly important for the use of HSV as compared to other virus vectors, as humans often harbour latent HSV. A vector virus carrying a transgene could interact with endogenous wild-type virus when used in gene therapy procedures. For this study we constructed four recombinant viruses containing marker genes to allow interactions between replication competent and disabled vector viruses to be explored. Recombination of viruses under replicating conditions was assessed in vitro and in vivo in the peripheral nervous system following the inoculation of combinations of viruses into the hindpaw of BALB/c mice and the examination of virus content in the dorsal root ganglia. Recombination between the viruses, when co-administered was found to occur such that transgene-bearing replication-competent viruses were generated only when the transgene was inserted at a non-essential site in the HSV genome. When the transgene was inserted into an essential site in the disabled virus, or when disabled and non-disabled virus were administered separately to the same site, transgene-bearing replication-competent recombinants were not observed. In the central nervous system, the ability of disabled, LacZ containing virus to reactivate latent replication-competent virus was examined. CNS latency was established by infecting BALB/c mice with a replication competent, GFP containing virus by the intra nasal route. After the establishment of latency, disabled virus was injected intra-cerebrally. Reactivation could not be detected as evidenced by a lack of GFP expression and replicating virus even though robust LacZ expression from the incoming vector virus could be detected. A similar lack of reactivation occurred when vector virus was inoculated into the footpad following the establishment of latency in the PNS by a replication competent virus. These experiments show (i) that insertion of the transgene in an essential site of the viral genome prevents its incorporation into an hazardous replication-competent recombinant derivative, indicating that non-homologous recombination between disabled and replication competent viruses does not occur at the level of sensitivity of the in vitro/in vivo assays used here, (ii) even homologous recombination in vivo only occurs at detectable levels when vector and replication competent virus are co-administered, and (iii) inoculation of vector HSV into the nervous system is very unlikely to reactivate latent wild-type virus that may be present.
{"title":"Examination of the potential interactions between herpes simplex virus vectors and replication-competent virus in vitro and in vivo","authors":"J. Smith, Suzanne Thomas, R. Coffin, D. Latchman","doi":"10.1163/156855803762295413","DOIUrl":"https://doi.org/10.1163/156855803762295413","url":null,"abstract":"In this paper, we have studied the potential interactions of replication incompetent herpes virus vectors with replication-competent virus both in vitro and in vivo. This might be thought to be particularly important for the use of HSV as compared to other virus vectors, as humans often harbour latent HSV. A vector virus carrying a transgene could interact with endogenous wild-type virus when used in gene therapy procedures. For this study we constructed four recombinant viruses containing marker genes to allow interactions between replication competent and disabled vector viruses to be explored. Recombination of viruses under replicating conditions was assessed in vitro and in vivo in the peripheral nervous system following the inoculation of combinations of viruses into the hindpaw of BALB/c mice and the examination of virus content in the dorsal root ganglia. Recombination between the viruses, when co-administered was found to occur such that transgene-bearing replication-competent viruses were generated only when the transgene was inserted at a non-essential site in the HSV genome. When the transgene was inserted into an essential site in the disabled virus, or when disabled and non-disabled virus were administered separately to the same site, transgene-bearing replication-competent recombinants were not observed. In the central nervous system, the ability of disabled, LacZ containing virus to reactivate latent replication-competent virus was examined. CNS latency was established by infecting BALB/c mice with a replication competent, GFP containing virus by the intra nasal route. After the establishment of latency, disabled virus was injected intra-cerebrally. Reactivation could not be detected as evidenced by a lack of GFP expression and replicating virus even though robust LacZ expression from the incoming vector virus could be detected. A similar lack of reactivation occurred when vector virus was inoculated into the footpad following the establishment of latency in the PNS by a replication competent virus. These experiments show (i) that insertion of the transgene in an essential site of the viral genome prevents its incorporation into an hazardous replication-competent recombinant derivative, indicating that non-homologous recombination between disabled and replication competent viruses does not occur at the level of sensitivity of the in vitro/in vivo assays used here, (ii) even homologous recombination in vivo only occurs at detectable levels when vector and replication competent virus are co-administered, and (iii) inoculation of vector HSV into the nervous system is very unlikely to reactivate latent wild-type virus that may be present.","PeriodicalId":93646,"journal":{"name":"Gene therapy and regulation","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2003-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1163/156855803762295413","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"64795146","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2003-03-01DOI: 10.1163/156855803762295431
K. Binley
The ability to deliver a therapeutic gene to the correct disease location and achieve expression at a relevant therapeutic level is important for the development of safe and potent gene-based therapies. Tissue-specific promoters restrict gene expression spatially in a cell-specific manner whereas pharmacologically responsive promoters can be controlled temporally through the administration of chemical repressors or activators. In some disease conditions such as cancer, it may be beneficial to restrict gene expression to a particular subset of diseased cells whilst preventing gene expression in the healthy portions of the tissue. This review focuses on the in vitro and in vivo studies which have fuelled the emergence of physiological regulation as an attractive means of spatially and temporally targeting gene expression. This review will centre on exploiting the physiological feature of hypoxia to target and control gene expression highlighting the recent success of hypoxia regulated gene therapy vectors in models of chronic anaemia and cancer.
{"title":"Exploiting hypoxia for targeted gene therapy","authors":"K. Binley","doi":"10.1163/156855803762295431","DOIUrl":"https://doi.org/10.1163/156855803762295431","url":null,"abstract":"The ability to deliver a therapeutic gene to the correct disease location and achieve expression at a relevant therapeutic level is important for the development of safe and potent gene-based therapies. Tissue-specific promoters restrict gene expression spatially in a cell-specific manner whereas pharmacologically responsive promoters can be controlled temporally through the administration of chemical repressors or activators. In some disease conditions such as cancer, it may be beneficial to restrict gene expression to a particular subset of diseased cells whilst preventing gene expression in the healthy portions of the tissue. This review focuses on the in vitro and in vivo studies which have fuelled the emergence of physiological regulation as an attractive means of spatially and temporally targeting gene expression. This review will centre on exploiting the physiological feature of hypoxia to target and control gene expression highlighting the recent success of hypoxia regulated gene therapy vectors in models of chronic anaemia and cancer.","PeriodicalId":93646,"journal":{"name":"Gene therapy and regulation","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2003-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1163/156855803762295431","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"64795337","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2001-12-01DOI: 10.1163/156855801760106993
J. Symes, P. Vale, R. Schatz, Douglas Losordo
Preclinical studies have documented that gene transfer of plasmid DNA encoding for secreted angiogenic cytokines such as vascular endothelial growth factor (VEGF) can induce therapeutic angiogenesis in animal models. Based on these studies, we have conducted 4 Phase I clinical trials of direct intramyocardial injection of VEGF plasmid DNA in patients with otherwise "inoperable" myocardial ischemia. In the first 2 trials, VEGF-1 (30 patients) or VEGF-2 (30 patients) plasmid DNA was administered through a mini-thoracotomy incision in an open label nonrandomized fashion as sole therapy. The majority of patients in both study groups experienced a significant reduction in angina frequency and improvement in Canadian Cardiovascular Society (CCS) angina class associated with objective evidence of improved myocardial perfusion documented by myocardial perfusion imaging and left ventricular electromechanical mapping. The subsequent development of a catheter-based approach to myocardial gene transfer has allowed us to conduct 2 randomized blinded trials of VEGF-2 gene transfer in a total of 25 patients. While a placebo effect was evident early post-treatment, VEGF-treated patients demonstrated significantly better results in terms of improvement in angina class and objective evidence of improvement in myocardial perfusion. These early Phase I trials confirm that direct myocardial gene transfer with VEGF is safe (overall 1 year mortality 3.5%) and should thus permit larger scale, appropriately randomized trials to be performed in order to more definitively evaluate its therapeutic efficacy.
{"title":"Clinical trials of myocardial VEGF gene transfer","authors":"J. Symes, P. Vale, R. Schatz, Douglas Losordo","doi":"10.1163/156855801760106993","DOIUrl":"https://doi.org/10.1163/156855801760106993","url":null,"abstract":"Preclinical studies have documented that gene transfer of plasmid DNA encoding for secreted angiogenic cytokines such as vascular endothelial growth factor (VEGF) can induce therapeutic angiogenesis in animal models. Based on these studies, we have conducted 4 Phase I clinical trials of direct intramyocardial injection of VEGF plasmid DNA in patients with otherwise \"inoperable\" myocardial ischemia. In the first 2 trials, VEGF-1 (30 patients) or VEGF-2 (30 patients) plasmid DNA was administered through a mini-thoracotomy incision in an open label nonrandomized fashion as sole therapy. The majority of patients in both study groups experienced a significant reduction in angina frequency and improvement in Canadian Cardiovascular Society (CCS) angina class associated with objective evidence of improved myocardial perfusion documented by myocardial perfusion imaging and left ventricular electromechanical mapping. The subsequent development of a catheter-based approach to myocardial gene transfer has allowed us to conduct 2 randomized blinded trials of VEGF-2 gene transfer in a total of 25 patients. While a placebo effect was evident early post-treatment, VEGF-treated patients demonstrated significantly better results in terms of improvement in angina class and objective evidence of improvement in myocardial perfusion. These early Phase I trials confirm that direct myocardial gene transfer with VEGF is safe (overall 1 year mortality 3.5%) and should thus permit larger scale, appropriately randomized trials to be performed in order to more definitively evaluate its therapeutic efficacy.","PeriodicalId":93646,"journal":{"name":"Gene therapy and regulation","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2001-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1163/156855801760106993","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"64794706","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2001-12-01DOI: 10.1163/156855801760107019
R. Morishita, M. Aoki, N. Hashiya, K. Yamasaki, H. Makino, Kohji Wakayama, J. Azuma, T. Ogihara
HGF is a mesenchyme-derived pleiotropic factor which regulates cell growth, cell motility, and morphogenesis of various types of cells, and is thus considered a humoral mediator of epithelial-mesenchymal interactions responsible for morphogenic tissue interactions during embryonic development and organogenesis. Although HGF was originally identified as a potent mitogen for hepatocytes, it has also been shown to be an angiogenic growth factor. Interestingly, the presence of its specific receptor, c-met, is observed in vascular cells and cardiac myocytes. In addition, among growth factors, the mitogenic action of HGF on human endothelial cells was most potent. Recent studies on animal models have demonstrated the potential application of HGF angiogenic gene therapy to treat cardiovascular diseases such as peripheral arterial disease (PAD), myocardial infarction, cerebrovascular disease and post-angioplastic restenosis. In this review, we present early results from an HGF gene therapy trial on six patients with PAD/Buerger disease and discuss the attractive promises of HGF gene transfer for cardiovascular diseases.
{"title":"Hepatocyte growth factor (HGF) angiogenic gene therapy: promises for cardiovascular diseases","authors":"R. Morishita, M. Aoki, N. Hashiya, K. Yamasaki, H. Makino, Kohji Wakayama, J. Azuma, T. Ogihara","doi":"10.1163/156855801760107019","DOIUrl":"https://doi.org/10.1163/156855801760107019","url":null,"abstract":"HGF is a mesenchyme-derived pleiotropic factor which regulates cell growth, cell motility, and morphogenesis of various types of cells, and is thus considered a humoral mediator of epithelial-mesenchymal interactions responsible for morphogenic tissue interactions during embryonic development and organogenesis. Although HGF was originally identified as a potent mitogen for hepatocytes, it has also been shown to be an angiogenic growth factor. Interestingly, the presence of its specific receptor, c-met, is observed in vascular cells and cardiac myocytes. In addition, among growth factors, the mitogenic action of HGF on human endothelial cells was most potent. Recent studies on animal models have demonstrated the potential application of HGF angiogenic gene therapy to treat cardiovascular diseases such as peripheral arterial disease (PAD), myocardial infarction, cerebrovascular disease and post-angioplastic restenosis. In this review, we present early results from an HGF gene therapy trial on six patients with PAD/Buerger disease and discuss the attractive promises of HGF gene transfer for cardiovascular diseases.","PeriodicalId":93646,"journal":{"name":"Gene therapy and regulation","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2001-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1163/156855801760107019","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"64794450","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2001-12-01DOI: 10.1163/156855801760107046
I. Kehat, L. Gepstein
The heart has relatively little regenerative capacity and hence any significant cell loss may lead to the development of heart failure. Cell therapy is emerging as a possible novel therapeutic approach for heart failure but is hampered by the paucity of cell sources for cardiomyocytes. Human embryonic stem (ES) cells are cell lines that were derived from the pre-implantation embryo and are capable of undifferentiated proliferation while retaining the capability to form derivatives of all three germ layers. The current review describes the properties of human ES cells and the derivation and properties of cardiomyocytes generated using this unique differentiating system. The possible applications in several research and clinical fields are described as well as the steps required to establish this tissue as a unique source for cell therapy procedures.
{"title":"Derivation and properties of human embryonic stem cell-derived cardiomyocytes","authors":"I. Kehat, L. Gepstein","doi":"10.1163/156855801760107046","DOIUrl":"https://doi.org/10.1163/156855801760107046","url":null,"abstract":"The heart has relatively little regenerative capacity and hence any significant cell loss may lead to the development of heart failure. Cell therapy is emerging as a possible novel therapeutic approach for heart failure but is hampered by the paucity of cell sources for cardiomyocytes. Human embryonic stem (ES) cells are cell lines that were derived from the pre-implantation embryo and are capable of undifferentiated proliferation while retaining the capability to form derivatives of all three germ layers. The current review describes the properties of human ES cells and the derivation and properties of cardiomyocytes generated using this unique differentiating system. The possible applications in several research and clinical fields are described as well as the steps required to establish this tissue as a unique source for cell therapy procedures.","PeriodicalId":93646,"journal":{"name":"Gene therapy and regulation","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2001-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1163/156855801760107046","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"64794911","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2001-12-01DOI: 10.1163/156855801760107028
T. Asahara, A. Kawamoto, C. Kalka, H. Masuda
Post-natal vasculogenesis has been considered to be involved in neovascularization of adult tissues, after bone marrow-derived endothelial progenitor cells (EPCs) were isolated from circulating mononuclear cells in peripheral blood and shown to incorporate into sites of physiological and pathological neovascularization and differentiate into mature endothelial cells (ECs). Following these findings, the various strategies to enhance neovascularization using EPCs have been researched, such as in vivo mobilization through exogenous administration of angiogenic factors, ex vivo culture expansion for cell transplantation or gene therapy. EPCs might have an attractive potential in therapeutic application for cardiovascular ischemic diseases as a novel cell strategy.
{"title":"Therapeutic potential of bone marrow-derived endothelial progenitor cells for cardiovascular ischemic diseases","authors":"T. Asahara, A. Kawamoto, C. Kalka, H. Masuda","doi":"10.1163/156855801760107028","DOIUrl":"https://doi.org/10.1163/156855801760107028","url":null,"abstract":"Post-natal vasculogenesis has been considered to be involved in neovascularization of adult tissues, after bone marrow-derived endothelial progenitor cells (EPCs) were isolated from circulating mononuclear cells in peripheral blood and shown to incorporate into sites of physiological and pathological neovascularization and differentiate into mature endothelial cells (ECs). Following these findings, the various strategies to enhance neovascularization using EPCs have been researched, such as in vivo mobilization through exogenous administration of angiogenic factors, ex vivo culture expansion for cell transplantation or gene therapy. EPCs might have an attractive potential in therapeutic application for cardiovascular ischemic diseases as a novel cell strategy.","PeriodicalId":93646,"journal":{"name":"Gene therapy and regulation","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2001-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1163/156855801760107028","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"64794566","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2001-12-01DOI: 10.1163/156855801760107055
E. Heber-Katz, J. Leferovich, Khamilia Bedelbaeva
The reaction of myocardium to injury is generally characterized by a tissue repair mechanism that is initially beneficial in maintaining tissue integrity but which ultimately has a debilitating effect on cardiac function. The components of this type of wound healing response in the mammalian heart have been the subject of extensive study. Far less understood, however, are the dynamics of inflammation, ECM remodeling and scar formation at the site of injury, as well as compensatory adaptations of adjacent uninjured myocardium such as angiogenesis, cadiomyocyte hypertrophy, and apoptosis in the regenerative response to a myocardial injury. Recent observations in the MRL strain of mice indicate a capacity to heal myocardial injury with a rapid resolution of granulation tissue and little scar tissue, an increase of cardiomyocyte BrdU incorporation and restoration of myocardial architecture/function. The role of stem cells in this process remains to be elucidated for MRL mice. This response to injury, however, suggests epimorphic regeneration rather than repair.
{"title":"Spontaneous heart regeneration in adult MRL mice after cryo-injury","authors":"E. Heber-Katz, J. Leferovich, Khamilia Bedelbaeva","doi":"10.1163/156855801760107055","DOIUrl":"https://doi.org/10.1163/156855801760107055","url":null,"abstract":"The reaction of myocardium to injury is generally characterized by a tissue repair mechanism that is initially beneficial in maintaining tissue integrity but which ultimately has a debilitating effect on cardiac function. The components of this type of wound healing response in the mammalian heart have been the subject of extensive study. Far less understood, however, are the dynamics of inflammation, ECM remodeling and scar formation at the site of injury, as well as compensatory adaptations of adjacent uninjured myocardium such as angiogenesis, cadiomyocyte hypertrophy, and apoptosis in the regenerative response to a myocardial injury. Recent observations in the MRL strain of mice indicate a capacity to heal myocardial injury with a rapid resolution of granulation tissue and little scar tissue, an increase of cardiomyocyte BrdU incorporation and restoration of myocardial architecture/function. The role of stem cells in this process remains to be elucidated for MRL mice. This response to injury, however, suggests epimorphic regeneration rather than repair.","PeriodicalId":93646,"journal":{"name":"Gene therapy and regulation","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2001-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1163/156855801760107055","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"64794476","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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