Pub Date : 2001-12-01DOI: 10.1163/156855801760107000
H. Hammond
An animal model of stress-induced regional myocardial ischemia was used to test the hypothesis that intracoronary delivery of an adenovirus encoding fibroblast growth factor Type 5 (FGF5) would result in improved blood flow, function and neovascularization in the ischemic region of the heart. These data indicated that this approach could be used safely and effectively in resolving regional myocardial ischemia in the ameroid model. Additional unpublished studies confirmed that a similar effect was achievable through use of other angiogenic transgenes, including FGF Type 4 (FGF4). The published studies using direct intracoronary delivery of adenovirus vectors to the heart is reviewed, with a focus on safety and efficacy. A multicenter Phase 1/Phase 2 clinical trial of intracoronary delivery of FGF4 in patients with angina pectoris was initiated under joint sponsorship of Collateral Therapeutics, Incorporated, Berlex Biosciences and Schering AG. Results of this trial are reviewed. The results of this blinded and randomized clinical trial provided data supporting the initiation of two large scale pivotal clinical trials — in the European Union and the US — of intracoronary delivery of adenovirus encoding FGF4 in treating patients with angina.
{"title":"Intracoronary gene transfer of fibroblast growth factor in experimental and clinical myocardial ischemia","authors":"H. Hammond","doi":"10.1163/156855801760107000","DOIUrl":"https://doi.org/10.1163/156855801760107000","url":null,"abstract":"An animal model of stress-induced regional myocardial ischemia was used to test the hypothesis that intracoronary delivery of an adenovirus encoding fibroblast growth factor Type 5 (FGF5) would result in improved blood flow, function and neovascularization in the ischemic region of the heart. These data indicated that this approach could be used safely and effectively in resolving regional myocardial ischemia in the ameroid model. Additional unpublished studies confirmed that a similar effect was achievable through use of other angiogenic transgenes, including FGF Type 4 (FGF4). The published studies using direct intracoronary delivery of adenovirus vectors to the heart is reviewed, with a focus on safety and efficacy. A multicenter Phase 1/Phase 2 clinical trial of intracoronary delivery of FGF4 in patients with angina pectoris was initiated under joint sponsorship of Collateral Therapeutics, Incorporated, Berlex Biosciences and Schering AG. Results of this trial are reviewed. The results of this blinded and randomized clinical trial provided data supporting the initiation of two large scale pivotal clinical trials — in the European Union and the US — of intracoronary delivery of adenovirus encoding FGF4 in treating patients with angina.","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/10.1163/156855801760107000","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"64794828","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: 10.1163/156855801760106984
I. Baumgartner
Therapeutic neovascularization is a new area in cardiovascular medicine that received a lot of attention during the past 4 years. Starting with Schumacher et al.'s trial (1998) using recombinant acidic fibroblast growth factor in a coronary setting of incompletely bypassed patients, and Baumgartner et al.'s gene therapy trial (1998) on a series of patients with critical limb ischemia unsuitable for conventional revascularization, there are presently several ongoing clinical trials. This review article focuses on the rationale of intramuscular gene therapy compared to recombinant protein administration, and gives an overview of the development of vascular endothelial growth factor (VEGF) gene therapy for peripheral arterial occlusive disease. An enormous effort has already been done; much is left to be done. The purpose must be to improve the life of patients.
{"title":"VEGF gene therapy for chronic critical limb ischemia","authors":"I. Baumgartner","doi":"10.1163/156855801760106984","DOIUrl":"https://doi.org/10.1163/156855801760106984","url":null,"abstract":"Therapeutic neovascularization is a new area in cardiovascular medicine that received a lot of attention during the past 4 years. Starting with Schumacher et al.'s trial (1998) using recombinant acidic fibroblast growth factor in a coronary setting of incompletely bypassed patients, and Baumgartner et al.'s gene therapy trial (1998) on a series of patients with critical limb ischemia unsuitable for conventional revascularization, there are presently several ongoing clinical trials. This review article focuses on the rationale of intramuscular gene therapy compared to recombinant protein administration, and gives an overview of the development of vascular endothelial growth factor (VEGF) gene therapy for peripheral arterial occlusive disease. An enormous effort has already been done; much is left to be done. The purpose must be to improve the life of patients.","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/10.1163/156855801760106984","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"64794388","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 : 2001-08-01DOI: 10.1163/156855800750367455
M. Wei, Jianping Wang, H. Xia, Qong Li, M. West
In vitro tests were performed to evaluate the suitability of primary vascular smooth muscle cells (VSMCs) as targets for retroviral vector-mediated atrial natriuretic peptide (ANP) gene transfer, tetracycline (Tet)-regulated expression of ANP transgene and microencapsulation for an ex vivo approach of ANP gene therapy. Rat ANP cDNA was thus cloned from rat atrial tissue, and then sub-cloned and packaged into retroviral vectors comprising standard or Tet-regulated gene expression cassettes. After high efficiency of marker LacZ gene transfer was demonstrated, the expression of immuno-reactive ANP (irANP) was analysed. Our results showed that, unlike non-transduced or LacZ-transduced control VSMCs, LrASN/PA317-transduced VSMCs secreted a significant amount of irANP (425 ± 50 pg/ml/105cells/24 hours, at peak). Importantly enough, LNtetPrtTFrA/PA317-transduced VSMCs were shown to exhibit efficient Tet-regulated expression of irANP with nontoxic concentrations of doxycycline. The biological activity of irANP produced by such engineered VSMCs was evidenced by cyclic GMP (cGMP) activation in LrASN-transduced VSMCs or in VSMCs exposed to conditioned media harvested from VSMCs secreting irANP. Further studies showed that transduced VSMCs synthesised and secreted more irANP than either rat primary endothelial cells, or skin fibroblasts or a transformed mouse fibroblast cell line. Most importantly, micro-encapsulation of engineered VSMCs in alginate did not alter Tet-regulated expression and long-term secretion of irANP. These results suggest that encapsulated engineered VSMCs may prove instrumental in longterm in vivo studies on ANP function and in the development of an ex vivo ANP gene therapy approach for disease states such as hypertension and congestive heart failure.
{"title":"Atrial natriuretic peptide gene transfer and regulated expression in primary vascular smooth muscle cells","authors":"M. Wei, Jianping Wang, H. Xia, Qong Li, M. West","doi":"10.1163/156855800750367455","DOIUrl":"https://doi.org/10.1163/156855800750367455","url":null,"abstract":"In vitro tests were performed to evaluate the suitability of primary vascular smooth muscle cells (VSMCs) as targets for retroviral vector-mediated atrial natriuretic peptide (ANP) gene transfer, tetracycline (Tet)-regulated expression of ANP transgene and microencapsulation for an ex vivo approach of ANP gene therapy. Rat ANP cDNA was thus cloned from rat atrial tissue, and then sub-cloned and packaged into retroviral vectors comprising standard or Tet-regulated gene expression cassettes. After high efficiency of marker LacZ gene transfer was demonstrated, the expression of immuno-reactive ANP (irANP) was analysed. Our results showed that, unlike non-transduced or LacZ-transduced control VSMCs, LrASN/PA317-transduced VSMCs secreted a significant amount of irANP (425 ± 50 pg/ml/105cells/24 hours, at peak). Importantly enough, LNtetPrtTFrA/PA317-transduced VSMCs were shown to exhibit efficient Tet-regulated expression of irANP with nontoxic concentrations of doxycycline. The biological activity of irANP produced by such engineered VSMCs was evidenced by cyclic GMP (cGMP) activation in LrASN-transduced VSMCs or in VSMCs exposed to conditioned media harvested from VSMCs secreting irANP. Further studies showed that transduced VSMCs synthesised and secreted more irANP than either rat primary endothelial cells, or skin fibroblasts or a transformed mouse fibroblast cell line. Most importantly, micro-encapsulation of engineered VSMCs in alginate did not alter Tet-regulated expression and long-term secretion of irANP. These results suggest that encapsulated engineered VSMCs may prove instrumental in longterm in vivo studies on ANP function and in the development of an ex vivo ANP gene therapy approach for disease states such as hypertension and congestive heart failure.","PeriodicalId":93646,"journal":{"name":"Gene therapy and regulation","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2001-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1163/156855800750367455","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"64794429","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 : 2001-08-01DOI: 10.1163/156855800750367437
D. Doolan, S. Hoffman
The technology of DNA vaccination has facilitated a revolutionary approach to developing an effective vaccine against complex pathogens, such as the Plasmodium spp. parasites that cause malaria, for which vaccines are not yet available. Their capacity to induce the CD8C T cell dependent CTL and IFN-γ responses which have heretoforth been difficult to induce by the more conventional vaccine technologies has now been established in mice, monkeys and humans, in malaria as well as in other disease systems. Despite the fact that the first and second generation DNA vaccines on their own have not been optimal, the potential of promising immune enhancement strategies for DNA vaccination suggests that the current efforts in this field are warranted.
{"title":"DNA-based vaccines against malaria and other diseases - from the laboratory to the clinic","authors":"D. Doolan, S. Hoffman","doi":"10.1163/156855800750367437","DOIUrl":"https://doi.org/10.1163/156855800750367437","url":null,"abstract":"The technology of DNA vaccination has facilitated a revolutionary approach to developing an effective vaccine against complex pathogens, such as the Plasmodium spp. parasites that cause malaria, for which vaccines are not yet available. Their capacity to induce the CD8C T cell dependent CTL and IFN-γ responses which have heretoforth been difficult to induce by the more conventional vaccine technologies has now been established in mice, monkeys and humans, in malaria as well as in other disease systems. Despite the fact that the first and second generation DNA vaccines on their own have not been optimal, the potential of promising immune enhancement strategies for DNA vaccination suggests that the current efforts in this field are warranted.","PeriodicalId":93646,"journal":{"name":"Gene therapy and regulation","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2001-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1163/156855800750367437","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"64794041","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 : 2001-08-01DOI: 10.1163/156855800750367428
R. Bertolotti
{"title":"Adult and embryonic-like stem cells: toward a major gene therapy breakthrough relying on autologous multipotent stem cells","authors":"R. Bertolotti","doi":"10.1163/156855800750367428","DOIUrl":"https://doi.org/10.1163/156855800750367428","url":null,"abstract":"","PeriodicalId":93646,"journal":{"name":"Gene therapy and regulation","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2001-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1163/156855800750367428","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"64793904","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 : 2001-08-01DOI: 10.1163/156855800750367446
M. Fussenegger
A variety of heterologous mammalian gene regulation systems are currently available for use in human gene therapy and tissue engineering. While individual gene regulation concepts vary significantly in key characteristics (regulation profiles, basal expression levels, human compatibility, immunogenicity, pharmacokinetics and bioavailability of the inducing agent), most of them are sufficiently elaborated to be considered for preclinical research and clinical gene therapy trials. As forefront basic research describes the cell-cycle, differentiation and apoptosis regulatory networks in mammalian cells as an increasingly complex highly interconnected globular regulon, it becomes apparent that many human diseases may originate from the slightest expression imbalances of key regulatory genes. We believe that successful gene therapy of such diseases will involve complex multi-(level) regulated multigene interventions based on a combination of several human-compatible heterologous gene regulation systems which enable optimal integration of therapeutic interventions into the cellular regulon as well as well-balanced expression of several therapeutic transgenes. Although such multiregulated multigene expression scenarios are still a vision, the following are already a scientific reality with cultured cells: (i) combination of two human compatible gene regulation systems to enable dual-regulated expression technology for independent adjustment of two different gene activities, (ii) regulatory cascades for muli-level regulated multigene interventions, and (iii) dual-autoregulated expression configurations for one-step installation of multi-level or reciprocal regulation of heterologous gene expression. This review summarizes pioneering efforts to establish dual-regulated expression technology, a strategy to combine more than one heterologous gene regulation system, to achieve difficult-to-attain cell phenotypes relevant for gene therapy, tissue engineering and basic research.
{"title":"Dual-regulated gene expression in mammalian cells, a novel approach to gene therapy","authors":"M. Fussenegger","doi":"10.1163/156855800750367446","DOIUrl":"https://doi.org/10.1163/156855800750367446","url":null,"abstract":"A variety of heterologous mammalian gene regulation systems are currently available for use in human gene therapy and tissue engineering. While individual gene regulation concepts vary significantly in key characteristics (regulation profiles, basal expression levels, human compatibility, immunogenicity, pharmacokinetics and bioavailability of the inducing agent), most of them are sufficiently elaborated to be considered for preclinical research and clinical gene therapy trials. As forefront basic research describes the cell-cycle, differentiation and apoptosis regulatory networks in mammalian cells as an increasingly complex highly interconnected globular regulon, it becomes apparent that many human diseases may originate from the slightest expression imbalances of key regulatory genes. We believe that successful gene therapy of such diseases will involve complex multi-(level) regulated multigene interventions based on a combination of several human-compatible heterologous gene regulation systems which enable optimal integration of therapeutic interventions into the cellular regulon as well as well-balanced expression of several therapeutic transgenes. Although such multiregulated multigene expression scenarios are still a vision, the following are already a scientific reality with cultured cells: (i) combination of two human compatible gene regulation systems to enable dual-regulated expression technology for independent adjustment of two different gene activities, (ii) regulatory cascades for muli-level regulated multigene interventions, and (iii) dual-autoregulated expression configurations for one-step installation of multi-level or reciprocal regulation of heterologous gene expression. This review summarizes pioneering efforts to establish dual-regulated expression technology, a strategy to combine more than one heterologous gene regulation system, to achieve difficult-to-attain cell phenotypes relevant for gene therapy, tissue engineering and basic research.","PeriodicalId":93646,"journal":{"name":"Gene therapy and regulation","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2001-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1163/156855800750367446","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"64794167","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}
M. Garcia-Blanco, M. Puttaraju, S. Mansfield, L. Mitchell
In human cells, the majority of the primary transcripts synthesized by RNA polymerase II contain interruptions in the coding sequences, which are known as introns. The spliceosome, a macromolecular enzyme, precisely and efficiently removes these introns in a process known as pre-messenger RNA splicing. This enzyme is also capable of recombining two RNAs in trans in a process known as spliceosome-mediated RNA trans -splicing. In this review, we highlight technologies that take advantage of this RNA trans -splicing, from gene therapy to genomics.
{"title":"Spliceosome-mediated RNA trans -splicing in gene therapy and genomics","authors":"M. Garcia-Blanco, M. Puttaraju, S. Mansfield, L. Mitchell","doi":"10.1163/156855800744584","DOIUrl":"https://doi.org/10.1163/156855800744584","url":null,"abstract":"In human cells, the majority of the primary transcripts synthesized by RNA polymerase II contain interruptions in the coding sequences, which are known as introns. The spliceosome, a macromolecular enzyme, precisely and efficiently removes these introns in a process known as pre-messenger RNA splicing. This enzyme is also capable of recombining two RNAs in trans in a process known as spliceosome-mediated RNA trans -splicing. In this review, we highlight technologies that take advantage of this RNA trans -splicing, from gene therapy to genomics.","PeriodicalId":93646,"journal":{"name":"Gene therapy and regulation","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2000-08-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1163/156855800744584","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"64793644","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}
One of the strategies used to alter the tropism of retroviral vectors involves the substitution of their envelope glycoproteins with those of other viruses. This results in the generation of pseudotype vector particles exhibiting properties that may be advantageous for gene transfer into certain human cells. Moreover, when the envelope glycoproteins from simian and human immunodeficiency viruses were used to pseudotype Murine Leukemia Virus (MLV), the resulting vectors mediated selective gene transfer into defined human cell types. Enhanced transduction efficacy and cell targeting via retroviral pseudotype vectors may be important prerequisites for future gene therapy applications.
{"title":"MLV-derived retroviral pseudotype vectors","authors":"J. Stitz, C. Buchholz, K. Cichutek","doi":"10.1163/156855800744601","DOIUrl":"https://doi.org/10.1163/156855800744601","url":null,"abstract":"One of the strategies used to alter the tropism of retroviral vectors involves the substitution of their envelope glycoproteins with those of other viruses. This results in the generation of pseudotype vector particles exhibiting properties that may be advantageous for gene transfer into certain human cells. Moreover, when the envelope glycoproteins from simian and human immunodeficiency viruses were used to pseudotype Murine Leukemia Virus (MLV), the resulting vectors mediated selective gene transfer into defined human cell types. Enhanced transduction efficacy and cell targeting via retroviral pseudotype vectors may be important prerequisites for future gene therapy applications.","PeriodicalId":93646,"journal":{"name":"Gene therapy and regulation","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2000-08-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1163/156855800744601","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"64793566","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}
A number of potent endogenous inhibitors targeting the tumor vasculature have recently been identified in tumor-bearing animals. Some of these angiogenesis inhibitors, including angiostatin, endostatin, and serpin antithrombin, seem to act specifically on the proliferating endothelial cells in the newly formed blood vessels. The discovery of these specific endothelial inhibitors not only increases our understanding of the functions of these molecules in the regulation of physiological and pathological angiogenesis, but also provides an important therapeutic strategy for cancer treatment. Several studies have demonstrated that antiangiogenic protein therapy with these inhibitors significantly suppresses the growth of a variety of tumors in mice. However, the dosages of these endogenous inhibitors used in animal studies seem to be too high for clinical trials. Other disadvantages of antiangiogenic protein therapy include repeated injections, prolonged treatment, potential transmission of toxins and infectious particles, and high cost for manufacturing large amounts of protein molecules. Thus, alternative approaches need to be developed in order to improve the antiangiogenic therapy with endogenous inhibitors. Perhaps gene therapy aimed to express these potent angiogenesis inhibitors in vivo is the most promising alternative approach that could transfer antiangiogenic therapy from animal experiments into the clinic. Although the development of this field is still in its early stages, several studies in animals have already provided evidence that this is a promising approach in the treatment of cancer. In this review article, I will discuss the therapeutic potentials of antiangiogenic molecules expressed from gene therapy vectors.
{"title":"Antiangiogenic gene therapy","authors":"Yihai Cao","doi":"10.1163/156855800744575","DOIUrl":"https://doi.org/10.1163/156855800744575","url":null,"abstract":"A number of potent endogenous inhibitors targeting the tumor vasculature have recently been identified in tumor-bearing animals. Some of these angiogenesis inhibitors, including angiostatin, endostatin, and serpin antithrombin, seem to act specifically on the proliferating endothelial cells in the newly formed blood vessels. The discovery of these specific endothelial inhibitors not only increases our understanding of the functions of these molecules in the regulation of physiological and pathological angiogenesis, but also provides an important therapeutic strategy for cancer treatment. Several studies have demonstrated that antiangiogenic protein therapy with these inhibitors significantly suppresses the growth of a variety of tumors in mice. However, the dosages of these endogenous inhibitors used in animal studies seem to be too high for clinical trials. Other disadvantages of antiangiogenic protein therapy include repeated injections, prolonged treatment, potential transmission of toxins and infectious particles, and high cost for manufacturing large amounts of protein molecules. Thus, alternative approaches need to be developed in order to improve the antiangiogenic therapy with endogenous inhibitors. Perhaps gene therapy aimed to express these potent angiogenesis inhibitors in vivo is the most promising alternative approach that could transfer antiangiogenic therapy from animal experiments into the clinic. Although the development of this field is still in its early stages, several studies in animals have already provided evidence that this is a promising approach in the treatment of cancer. In this review article, I will discuss the therapeutic potentials of antiangiogenic molecules expressed from gene therapy vectors.","PeriodicalId":93646,"journal":{"name":"Gene therapy and regulation","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2000-08-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1163/156855800744575","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"64793470","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}
In hepatic fibrosis, the connective tissue biomatrix of the liver changes from the normal matrix, rich in basement membrane collagens, to a matrix enriched in interstitial fibrillar collagens. Type I collagen is the predominant component of thick fibrous bands found in matrix in advanced fibrosis. The aim of the current research was to determine whether a therapeutic approach could be developed that would specifically target collagen-producing cells to reduce the synthesis and accumulation of type I collagen. Antisense DNA oligonucleotides directed against specific sequences within α1(I) and α2(I) mRNA of type I procollagen were complexed to a cell-specific carrier, and screened for their effectiveness in reducing α1(I) and α2(I) mRNA levels. Two antisense DNA oligonucleotides delivered by the carrier were found to be most effective in reducing α1(I) and α2(I) mRNA and total collagen accumulation in the cells, but had no effect on reducing β-actin mRNA in the same cells. At similar concentrations, free antisense DNA oligonucleotides were not effective in inhibiting collagen synthesis, and/or in decreasing cellular concentrations of α1(I) or α2(I) mRNA. Collagen synthesis and mRNA levels in cells lacking receptors that recognize the carrier protein were not changed after treatment with complexed antisense DNA. The results indicate that antisense oligonucleotides can be targeted to cell types, and were effective inhibiting collagen synthesis in those cells.
{"title":"Targeted inhibition of type I procollagen synthesis by antisense DNA oligonucleotides","authors":"Catherine H. Wu, C. Walton, George Y Wu","doi":"10.1163/156855800744610","DOIUrl":"https://doi.org/10.1163/156855800744610","url":null,"abstract":"In hepatic fibrosis, the connective tissue biomatrix of the liver changes from the normal matrix, rich in basement membrane collagens, to a matrix enriched in interstitial fibrillar collagens. Type I collagen is the predominant component of thick fibrous bands found in matrix in advanced fibrosis. The aim of the current research was to determine whether a therapeutic approach could be developed that would specifically target collagen-producing cells to reduce the synthesis and accumulation of type I collagen. Antisense DNA oligonucleotides directed against specific sequences within α1(I) and α2(I) mRNA of type I procollagen were complexed to a cell-specific carrier, and screened for their effectiveness in reducing α1(I) and α2(I) mRNA levels. Two antisense DNA oligonucleotides delivered by the carrier were found to be most effective in reducing α1(I) and α2(I) mRNA and total collagen accumulation in the cells, but had no effect on reducing β-actin mRNA in the same cells. At similar concentrations, free antisense DNA oligonucleotides were not effective in inhibiting collagen synthesis, and/or in decreasing cellular concentrations of α1(I) or α2(I) mRNA. Collagen synthesis and mRNA levels in cells lacking receptors that recognize the carrier protein were not changed after treatment with complexed antisense DNA. The results indicate that antisense oligonucleotides can be targeted to cell types, and were effective inhibiting collagen synthesis in those cells.","PeriodicalId":93646,"journal":{"name":"Gene therapy and regulation","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2000-08-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1163/156855800744610","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"64793686","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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