Pub Date : 2004-08-01DOI: 10.1163/1568558042457479
A. S. Hirsh, J. Joung
Gene therapy reagents such as artificial transcription factors and site-specific endonucleases require "made-to-order" DNA-binding domains with high affinity and specificity for novel target sequences. Cys2His2 zinc finger proteins are the best understood and most commonly used framework for design and selection of such domains. Though a number of design strategies have been described in the literature, they vary significantly in their reliability and ease of execution. This situation has made it difficult for the non-specialist researcher to know how best to construct zinc finger proteins for their application of interest. This article reviews the current state of the technology and its limitations, and discusses prospects for improving our ability to make customized DNA-binding modules.
{"title":"Engineered Cys 2 His 2 zinc finger DNA-binding domains","authors":"A. S. Hirsh, J. Joung","doi":"10.1163/1568558042457479","DOIUrl":"https://doi.org/10.1163/1568558042457479","url":null,"abstract":"Gene therapy reagents such as artificial transcription factors and site-specific endonucleases require \"made-to-order\" DNA-binding domains with high affinity and specificity for novel target sequences. Cys2His2 zinc finger proteins are the best understood and most commonly used framework for design and selection of such domains. Though a number of design strategies have been described in the literature, they vary significantly in their reliability and ease of execution. This situation has made it difficult for the non-specialist researcher to know how best to construct zinc finger proteins for their application of interest. This article reviews the current state of the technology and its limitations, and discusses prospects for improving our ability to make customized DNA-binding modules.","PeriodicalId":93646,"journal":{"name":"Gene therapy and regulation","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2004-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1163/1568558042457479","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"64795459","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 : 2004-08-01DOI: 10.1163/1568558042457460
Kyoji Ueda, Y. Hanazono, N. Ageyama, H. Shibata, Satoko Ogata, Y. Ueda, T. Tabata, S. Ikehara, M. Taniwaki, M. Hasegawa, K. Terao, K. Ozawa
It has recently been reported that bone marrow cells can efficiently engraft without marrow conditioning when implanted directly into the bone marrow cavity (intra-bone marrow transplantation, iBMT) in mice. We have successfully examined the efficacy of autologous iBMT in a cynomolgus monkey model in conjuction with an in vivo expansion of transplanted cells by a selective amplifier transgene (Ueda et al., 2004) and provide here the detailed parameters of our iBMT method. We injected retrovirally-marked autologous CD34+ cells directly into the non-conditioned marrow cavity of the femur and humerus after gently irrigating the cavity with saline. This transplant procedure was safely performed without pulmonary embolism. Gene-marked cells were not detectable in the peripheral blood at one hour and one day after iBMT as assessed by sensitive PCR, indicating that iBMT hardly generated a systemic delivery of transplanted cells. On the other hand, 2 to 30% of clonogenic hematopoietic colonies produced from the implanted marrow were gene-marked at 6–12 months after iBMT. Our iBMT method for non-human primates is thus discussed in terms of long-lived hematopoietic stem/progenitor cells, bone marrow niche and long-term engraftment after iBMT without myeloablative conditioning.
最近有报道称，将骨髓细胞直接植入小鼠骨髓腔(intra-bone marrow transplantation, iBMT)，无需骨髓调节即可有效植入。我们已经成功地在食蟹猴模型中检测了自体iBMT的有效性，并通过转基因选择放大器在体内扩增移植细胞(Ueda et al.， 2004)，并在这里提供了我们的iBMT方法的详细参数。我们将逆转录病毒标记的自体CD34+细胞直接注射到股骨和肱骨的非条件骨髓腔内，然后用盐水轻轻冲洗腔内。移植手术是安全的，没有肺栓塞。通过敏感PCR评估，在iBMT后1小时和1天的外周血中未检测到基因标记的细胞，表明iBMT几乎没有产生移植细胞的全身递送。另一方面，移植骨髓产生的2 - 30%的克隆造血菌落在iBMT后6-12个月被标记为基因。因此，我们对非人灵长类动物的iBMT方法进行了讨论，包括长期存活的造血干细胞/祖细胞、骨髓生态位和iBMT后无清髓条件的长期植入。
{"title":"Method — Intra-bone marrow transplantation of hematopoietic stem cells in non-human primates: long-term engraftment without conditioning","authors":"Kyoji Ueda, Y. Hanazono, N. Ageyama, H. Shibata, Satoko Ogata, Y. Ueda, T. Tabata, S. Ikehara, M. Taniwaki, M. Hasegawa, K. Terao, K. Ozawa","doi":"10.1163/1568558042457460","DOIUrl":"https://doi.org/10.1163/1568558042457460","url":null,"abstract":"It has recently been reported that bone marrow cells can efficiently engraft without marrow conditioning when implanted directly into the bone marrow cavity (intra-bone marrow transplantation, iBMT) in mice. We have successfully examined the efficacy of autologous iBMT in a cynomolgus monkey model in conjuction with an in vivo expansion of transplanted cells by a selective amplifier transgene (Ueda et al., 2004) and provide here the detailed parameters of our iBMT method. We injected retrovirally-marked autologous CD34+ cells directly into the non-conditioned marrow cavity of the femur and humerus after gently irrigating the cavity with saline. This transplant procedure was safely performed without pulmonary embolism. Gene-marked cells were not detectable in the peripheral blood at one hour and one day after iBMT as assessed by sensitive PCR, indicating that iBMT hardly generated a systemic delivery of transplanted cells. On the other hand, 2 to 30% of clonogenic hematopoietic colonies produced from the implanted marrow were gene-marked at 6–12 months after iBMT. Our iBMT method for non-human primates is thus discussed in terms of long-lived hematopoietic stem/progenitor cells, bone marrow niche and long-term engraftment after iBMT without myeloablative conditioning.","PeriodicalId":93646,"journal":{"name":"Gene therapy and regulation","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2004-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1163/1568558042457460","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"64795901","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 : 2004-08-01DOI: 10.1163/1568558042457488
K. Pulkkanen, H. Sallinen, K. Tyynelä, S. Ylä-Herttuala
By now, we have seen the dawn of the clinical era of cancer gene therapy. An enormous amount of biological knowledge and experimental data were gathered during a short period of pre-clinical years, and numerous applications proceeded into human trials. The first clinical years have taught us hard lessons, and often forced us to pull back and choose new and alternative courses. However, they have also given sufficient promise that cancer gene therapy will establish its foothold in clinical cancer management in the future. More importantly, pioneering trials have helped us to put things into the right perspective; clinical advance will likely emerge from the combined use of gene therapy with traditional therapies. It also appears that no cancer gene therapy approach can be applied universally, but rather we will see treatments that show effectiveness in only a particular type of tumor. For the last but not the least — while researchers are likely to increase their efforts to find more efficient solutions — scientific rigor and study design should not be jeopardised by potential pressures from investors and drug companies. In this review, we summarize highlights in i) the current approaches and genetic treatment modalities that are available based on pre-clinical studies, and ii) the status of cancer gene therapy in current clinical oncology, with special reference to those cancers where gene therapy has proceeded to human trials.
{"title":"Cancer gene therapy — current status in the clinics","authors":"K. Pulkkanen, H. Sallinen, K. Tyynelä, S. Ylä-Herttuala","doi":"10.1163/1568558042457488","DOIUrl":"https://doi.org/10.1163/1568558042457488","url":null,"abstract":"By now, we have seen the dawn of the clinical era of cancer gene therapy. An enormous amount of biological knowledge and experimental data were gathered during a short period of pre-clinical years, and numerous applications proceeded into human trials. The first clinical years have taught us hard lessons, and often forced us to pull back and choose new and alternative courses. However, they have also given sufficient promise that cancer gene therapy will establish its foothold in clinical cancer management in the future. More importantly, pioneering trials have helped us to put things into the right perspective; clinical advance will likely emerge from the combined use of gene therapy with traditional therapies. It also appears that no cancer gene therapy approach can be applied universally, but rather we will see treatments that show effectiveness in only a particular type of tumor. For the last but not the least — while researchers are likely to increase their efforts to find more efficient solutions — scientific rigor and study design should not be jeopardised by potential pressures from investors and drug companies. In this review, we summarize highlights in i) the current approaches and genetic treatment modalities that are available based on pre-clinical studies, and ii) the status of cancer gene therapy in current clinical oncology, with special reference to those cancers where gene therapy has proceeded to human trials.","PeriodicalId":93646,"journal":{"name":"Gene therapy and regulation","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2004-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1163/1568558042457488","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"64795794","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 : 2003-08-01DOI: 10.1163/156855803322664628
S. Loiler, T. Flotte
The advent of new serotypes of adeno-associated viruses (AAV) has expanded the tissue tropism and allowed clinically relevant numbers of cells to be transduced by rAAV vectors. The two most significant obstacles in the way of future gene therapies with AAV vectors are tissue/target cell specificity and sitel-specific integration. Other reviews contained in this issue will look at advances in site-specific integration. This review will focus on the latest developments in AAV vector tropism and tissue/ target cell specificity.
{"title":"Alternative capsid strategies for targeting recombinant AAV gene therapy","authors":"S. Loiler, T. Flotte","doi":"10.1163/156855803322664628","DOIUrl":"https://doi.org/10.1163/156855803322664628","url":null,"abstract":"The advent of new serotypes of adeno-associated viruses (AAV) has expanded the tissue tropism and allowed clinically relevant numbers of cells to be transduced by rAAV vectors. The two most significant obstacles in the way of future gene therapies with AAV vectors are tissue/target cell specificity and sitel-specific integration. Other reviews contained in this issue will look at advances in site-specific integration. This review will focus on the latest developments in AAV vector tropism and tissue/ target cell specificity.","PeriodicalId":93646,"journal":{"name":"Gene therapy and regulation","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2003-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1163/156855803322664628","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"64795417","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 : 2003-08-01DOI: 10.1163/156855803322664592
R. Bertolotti
{"title":"Stem cell gene therapy: breakthrough culminating in combination of ex vivo protocols with transient topical gene therapy","authors":"R. Bertolotti","doi":"10.1163/156855803322664592","DOIUrl":"https://doi.org/10.1163/156855803322664592","url":null,"abstract":"","PeriodicalId":93646,"journal":{"name":"Gene therapy and regulation","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2003-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1163/156855803322664592","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"64795062","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 : 2003-08-01DOI: 10.1163/156855803322664619
N. A. Allen, R. Samulski
The field of gene therapy is progressing with continuous improvements in vector development and regulated transgene expression. These contributions will eventually allow matching the best vector system for the specific disease indication. However one essential component of gene delivery that may be more retractile to this type of development and critical to all methods of delivery relates to the molecular fate of the therapeutic nucleic acids after successful in vivo delivery. The importance of this was illustrated in the clinic when leukemias were a result of insertional mutagenesis after attempting to achieve long-term gene expression via retroviral vectors. These results echo the fact that safer methods of achieving long-term gene expression are needed for successful gene therapy vectors. One mechanism to be considered to obtain the desired gene therapy results of safe long-term gene expression is that of the targeted integration. A number of recombination systems both eukaryotic and prokaryotic are being evaluated for this purpose. Included in this list is the targeted recombination of the Adeno-Associated Virus (AAV). AAV is a unique virus in that it can establish latency in human cells by targeting the recombination of its genome to a specific locus on chromosome 19. This review discusses the current understanding of the cis and trans requirements for AAV site-specific recombination and the mechanism involved. The risks of AAV-based site-specific recombination are also addressed as well as the current state of gene delivery vectors based on the site-specific integration of AAV.
{"title":"The pros and cons of using the mechanism of AAV site-specific recombination in gene delivery","authors":"N. A. Allen, R. Samulski","doi":"10.1163/156855803322664619","DOIUrl":"https://doi.org/10.1163/156855803322664619","url":null,"abstract":"The field of gene therapy is progressing with continuous improvements in vector development and regulated transgene expression. These contributions will eventually allow matching the best vector system for the specific disease indication. However one essential component of gene delivery that may be more retractile to this type of development and critical to all methods of delivery relates to the molecular fate of the therapeutic nucleic acids after successful in vivo delivery. The importance of this was illustrated in the clinic when leukemias were a result of insertional mutagenesis after attempting to achieve long-term gene expression via retroviral vectors. These results echo the fact that safer methods of achieving long-term gene expression are needed for successful gene therapy vectors. One mechanism to be considered to obtain the desired gene therapy results of safe long-term gene expression is that of the targeted integration. A number of recombination systems both eukaryotic and prokaryotic are being evaluated for this purpose. Included in this list is the targeted recombination of the Adeno-Associated Virus (AAV). AAV is a unique virus in that it can establish latency in human cells by targeting the recombination of its genome to a specific locus on chromosome 19. This review discusses the current understanding of the cis and trans requirements for AAV site-specific recombination and the mechanism involved. The risks of AAV-based site-specific recombination are also addressed as well as the current state of gene delivery vectors based on the site-specific integration of AAV.","PeriodicalId":93646,"journal":{"name":"Gene therapy and regulation","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2003-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1163/156855803322664619","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"64795351","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 : 2003-08-01DOI: 10.1163/156855803322664600
Eric C. Olivares, M. Calos
For most genetic disorders, long-term correction is necessary. Integration of a therapeutic gene into a patient's genome is an obvious route to achieving such permanent correction. Several technologies have been applied to the goal of achieving integration, including viruses and transposases. While these techniques are effective at some level, they each have drawbacks that can be improved upon. A novel integration system based on a phage integrase can address some of the previous limitations. The integrase from the Streptomyces bacteriophage C31 catalyzes site-specific, unidirectional integration into the genomes of higher eukaryotes. This integrase has the ability to recognize a limited number of native genomic sequences and integrate introduced plasmid DNA into them. These native sequences, termed pseudo att sites, resemble the wild-type phage attachment site enough to support integrase-mediated integration. Molecular evolution holds the promise of creating custom integrases that preferentially recombine at particular pseudo att sites. Furthermore, the system has no apparent size limit on carrying capacity. These features make the C31 integrase system extremely appealing for gene therapy applications. The system has been successfully employed in several model gene therapy studies to date. Here we review the development of this novel integration system and its current and potential applications to gene therapy.
{"title":"Phage φC31 integrase-mediated site-specific integration for gene therapy","authors":"Eric C. Olivares, M. Calos","doi":"10.1163/156855803322664600","DOIUrl":"https://doi.org/10.1163/156855803322664600","url":null,"abstract":"For most genetic disorders, long-term correction is necessary. Integration of a therapeutic gene into a patient's genome is an obvious route to achieving such permanent correction. Several technologies have been applied to the goal of achieving integration, including viruses and transposases. While these techniques are effective at some level, they each have drawbacks that can be improved upon. A novel integration system based on a phage integrase can address some of the previous limitations. The integrase from the Streptomyces bacteriophage C31 catalyzes site-specific, unidirectional integration into the genomes of higher eukaryotes. This integrase has the ability to recognize a limited number of native genomic sequences and integrate introduced plasmid DNA into them. These native sequences, termed pseudo att sites, resemble the wild-type phage attachment site enough to support integrase-mediated integration. Molecular evolution holds the promise of creating custom integrases that preferentially recombine at particular pseudo att sites. Furthermore, the system has no apparent size limit on carrying capacity. These features make the C31 integrase system extremely appealing for gene therapy applications. The system has been successfully employed in several model gene therapy studies to date. Here we review the development of this novel integration system and its current and potential applications to gene therapy.","PeriodicalId":93646,"journal":{"name":"Gene therapy and regulation","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2003-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1163/156855803322664600","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"64795143","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 : 2003-08-01DOI: 10.1163/156855803322664637
S. Rivella, L. Lisowski, M. Sadelain
The β-thalassemias and sickle cell disease are severe congenital anemias that are caused by the defective synthesis of the β chain of hemoglobin. Allogeneic hematopoietic stem cell (HSC) transplantation is curative, but this therapeutic option is not available to the majority of patients. The transfer of a regulated β-globin gene in autologous HCSs thus represents a highly attractive alternative treatment. This strategy, simple in principle, raises major challenges in terms of controlling transgene expression, which ideally should be erythroid-specific, differentiation stage-restricted, elevated, position-independent, and sustained over time. Using lentiviral vectors, we recently demonstrated that an optimized combination of proximal and distal transcriptional control elements permits lineage-specific and elevated expression of the β-globin gene, resulting in therapeutic hemoglobin production and correction of anemia in β-thalassemic mice. Several groups have now confirmed and extended these findings in various mouse models of severe hemoglobinopathies, thus generating enthusiasm for a genetic treatment based on globin gene transfer. Furthermore, globin vectors provide a paradigm for improving vector safety by restricting transgene expression to the differentiated progeny within a single lineage, thereby reducing the risk of activating oncogenes in hematopoietic progenitors. Here we review the principles underlying the genesis of regulated vectors for stem cell therapy.
{"title":"Globin gene transfer: a paradigm for transgene regulation and vector safety","authors":"S. Rivella, L. Lisowski, M. Sadelain","doi":"10.1163/156855803322664637","DOIUrl":"https://doi.org/10.1163/156855803322664637","url":null,"abstract":"The β-thalassemias and sickle cell disease are severe congenital anemias that are caused by the defective synthesis of the β chain of hemoglobin. Allogeneic hematopoietic stem cell (HSC) transplantation is curative, but this therapeutic option is not available to the majority of patients. The transfer of a regulated β-globin gene in autologous HCSs thus represents a highly attractive alternative treatment. This strategy, simple in principle, raises major challenges in terms of controlling transgene expression, which ideally should be erythroid-specific, differentiation stage-restricted, elevated, position-independent, and sustained over time. Using lentiviral vectors, we recently demonstrated that an optimized combination of proximal and distal transcriptional control elements permits lineage-specific and elevated expression of the β-globin gene, resulting in therapeutic hemoglobin production and correction of anemia in β-thalassemic mice. Several groups have now confirmed and extended these findings in various mouse models of severe hemoglobinopathies, thus generating enthusiasm for a genetic treatment based on globin gene transfer. Furthermore, globin vectors provide a paradigm for improving vector safety by restricting transgene expression to the differentiated progeny within a single lineage, thereby reducing the risk of activating oncogenes in hematopoietic progenitors. Here we review the principles underlying the genesis of regulated vectors for stem cell therapy.","PeriodicalId":93646,"journal":{"name":"Gene therapy and regulation","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2003-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1163/156855803322664637","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"64795432","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: 10.1163/156855803762295422
A. Jacobs, A. Winkeler, C. Dittmar, S. Vollmar, K. Wienhard, J. Voges, W. Heiss
A phase I/II clinical trial of gene therapy for recurrent glioblastoma has been initiated in which non-invasive monitoring of the critical steps is performed by a combination of standard clinical positron-emission tomography (PET) for metabolic identification/evaluation of malignant lesions with pioneering PET imaging of HSV-1- tk transgene expression. Localization, characterization and trial follow-up of target tumors rely on multitracer PET brain imaging probed with radiolabeled 2-[ 18 F]fluoro-2-deoxy-D-glucose (FDG), methyl-[ 11 C]-L-methionine (MET) and 3´-deoxy-3´-[ 18 F]fluoro-L-thymidine (FLT) while the location, magnitude and duration of therapeutic HSV-1- tk transgene expression is monitored with 2´-fluoro-2´-deoxy-1-β-D-arabinofuranosyl-5-[ 124 I]iod ([ 124 I]-FIAU). In addition, in order to improve the distribution of the vector within the tumor tissue, a stereotactically guided convection-enhanced delivery (CED) protocol was devised in which the transfection of the HSV-1- tk gene into brain tumor cells is mediated by a liposome-DNA complex. Preliminary findings on a first group of five patients demonstrated that FIAU-PET imaging of HSV-1- tk expression in patients with glioblastoma is feasible and that vector-mediated gene expression may predict the therapeutic effect of ganciclovir prodrug activation. In addition, they showed that integration of magnetic resonance (MR) and PET imaging data into a 3D stereotaxic coordinate system results in an efficient non-invasive spatio-temporal monitoring of a brain gene therapy trial. Such a non-invasive imaging provides the means to identify potential critical parameters and to implement dosing/ routing adjustments that will have a critical impact on the development of standardized gene therapy protocols.
{"title":"Positron-emission-tomography monitoring of anti-glioblastoma HSV-1-tk gene therapy","authors":"A. Jacobs, A. Winkeler, C. Dittmar, S. Vollmar, K. Wienhard, J. Voges, W. Heiss","doi":"10.1163/156855803762295422","DOIUrl":"https://doi.org/10.1163/156855803762295422","url":null,"abstract":"A phase I/II clinical trial of gene therapy for recurrent glioblastoma has been initiated in which non-invasive monitoring of the critical steps is performed by a combination of standard clinical positron-emission tomography (PET) for metabolic identification/evaluation of malignant lesions with pioneering PET imaging of HSV-1- tk transgene expression. Localization, characterization and trial follow-up of target tumors rely on multitracer PET brain imaging probed with radiolabeled 2-[ 18 F]fluoro-2-deoxy-D-glucose (FDG), methyl-[ 11 C]-L-methionine (MET) and 3´-deoxy-3´-[ 18 F]fluoro-L-thymidine (FLT) while the location, magnitude and duration of therapeutic HSV-1- tk transgene expression is monitored with 2´-fluoro-2´-deoxy-1-β-D-arabinofuranosyl-5-[ 124 I]iod ([ 124 I]-FIAU). In addition, in order to improve the distribution of the vector within the tumor tissue, a stereotactically guided convection-enhanced delivery (CED) protocol was devised in which the transfection of the HSV-1- tk gene into brain tumor cells is mediated by a liposome-DNA complex. Preliminary findings on a first group of five patients demonstrated that FIAU-PET imaging of HSV-1- tk expression in patients with glioblastoma is feasible and that vector-mediated gene expression may predict the therapeutic effect of ganciclovir prodrug activation. In addition, they showed that integration of magnetic resonance (MR) and PET imaging data into a 3D stereotaxic coordinate system results in an efficient non-invasive spatio-temporal monitoring of a brain gene therapy trial. Such a non-invasive imaging provides the means to identify potential critical parameters and to implement dosing/ routing adjustments that will have a critical impact on the development of standardized gene therapy protocols.","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/10.1163/156855803762295422","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"64795216","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: 10.1163/156855803762295440
P. Davis, A. Ziady
The airway epithelium has been an important target for gene therapy in the last decade. This route of administration is readily accessible and the airway is affected by a number of disorders for which gene therapy may be useful. Viral vectors were first used to transfer genes to the airway of cystic fibrosis patients a decade ago. However, in vivo results have been disappointing, with substantial inflammation and toxicity occurring at doses that do not produce significant gene transfer. More recently researchers have used plasmid DNA based technologies as an alternative. DNA alone can transfect cells in culture and in vivo, though it is inefficient. Complexing DNA with cationic molecules improves efficiency. Nonviral gene transfer with lipid-DNA complexes has showed promise in cell culture and animals, but is much less efficacious in humans, and inflammation occurs at doses below those required for therapeutic effect. Trans-nuclear membrane delivery appears to be the limiting factor of current liposome-DNA complex efficiency in vivo. DNA compacted with polycations has also been used with good success in animal models, with minimal toxicity and is currently being tested in human trials. In this review, we will discuss the advances and limitations of these nonviral vectors in gene delivery, particularly to airway epithelial cells.
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