ISSN:1043-0342    

DOI:10.1089/hum.1993.4.1-1

出版商:Mary Ann Liebert, Inc.    

年代:1993

数据来源: MAL

摘要:

ABSTRACTPrimary skin fibroblasts are an attractive target tissue for retroviral-mediated gene therapy; however, transient expression of therapeutic genes has been a recurrent problem in several rodent models. The gradual decrease in gene expression could be tissue or species specific. To investigate the phenomenon further, human adenosine deaminase (ADA) expression was monitored in genetically modified skin fibroblasts transplanted in beagle dogs. In culture, transduced canine fibroblasts expressed high levels of human ADA activity (33.6moles adenosine metabolized per hour per milligram of soluble protein) in comparison to canine ADA in untreated control cells (1.3mole/hrmg protein) and for 2 weeks following transplantation, the graft contained up to four-fold more enzyme activity from human ADA than the endogenous canine enzyme. However, by 10 weeks, human ADA expression was undetectable. At the time when human ADA expression was greatly reduced, DNA analysis showed the presence of vector sequences. These results directly parallel those observed in rodent models and suggest retroviral vector inactivation is a tissue-specific rather than species-specific mechanism.

ISSN:1043-0342    

DOI:10.1089/hum.1993.4.1-3

出版商:Mary Ann Liebert, Inc.    

年代:1993

数据来源: MAL

摘要:

ABSTRACTAdenosine deaminase (ADA) deficiency, a rare autosomal recessive disorder, is an ideal candidate for gene replacement therapy. By means of co-cultivation with a retroviral vector-producing cell line, we have demonstrated efficient transfer and expression of the human ADA gene into human primitive hematopoietic progenitors. At 6 weeks post-transduction in myeloid long-term bone marrow culture, approximately 50% of the clonogenic progenitors were transduced by the provirus, with ADA expression detected in 30% of transduced colonies. The ADA activity increased by 3.7-fold in the nonadherent fraction of transduced bone marrow after 9 weeks. We have also achieved efficient transduction by retroviral supernatant of normal and ADA-deficient bone marrow cells that were allowed to establish a stromal layer in long-term culture, indicating the feasibility of proceeding with attempts to perform stem cell gene therapy on patients with ADA deficiency.

ISSN:1043-0342    

DOI:10.1089/hum.1993.4.1-9

出版商:Mary Ann Liebert, Inc.    

年代:1993

数据来源: MAL

摘要:

ABSTRACTAdenoviruspolylysineDNA complexes were evaluated for their capacity to accomplish directin vivogene transfer to airway epithelium employing a rodent model. Binary complexes containing transferrin or adenovirus, or combination complexes containing both transferrin and adenovirus, were evaluated. The highestin vitrogene transfer efficiency in primary cultures of airway epithelial cells was accomplished by the combination complexes. This result was paralleledin vivo. Transient gene expression of up to 1 week was observed with localization of the transduced cells to the region of the small airways. These results establish the feasibility of this type of approach for gene therapy applications.

ISSN:1043-0342    

DOI:10.1089/hum.1993.4.1-17

出版商:Mary Ann Liebert, Inc.    

年代:1993

数据来源: MAL

摘要:

ABSTRACTThe CD18 gene encodes the2-subunit of leukocyte integrins, and mutations in this gene cause extreme host susceptibility to bacterial and fungal infection. Because expression of CD18 is restricted to bone marrow-derived cells, this disorder is considered an excellent candidate for somatic gene therapy utilizingex vivoinfection of bone marrow stem cells. We have constructed a retroviral vector expressing CD18 with the Moloney murine leukemia virus (Mo-MLV) long terminal repeat (LTR) as the promoter, and high-titer ecotropic and amphotropic producer cell lines were isolated using the GPE-86 and GPenvAM12 safe packaging cell lines. Infection of CD18-deficient lymphoblasts resulted both in expression of immunodetectable CD18 at 3540% of normal levels on 5560% of cells and in functional restoration of CD18-dependent aggregation. All of 16 mice transplanted with syngeneic bone marrow infected with the CD18 retrovirus expressed human CD18 on 1736% of granulocytes at 2 weeks after transplantation, and expression was appropriately up-regulated in response to stimulation with zymosan-activated serum. This recombinant retrovirus should prove useful for further studies of somatic gene therapy for CD18 deficiency.

ISSN:1043-0342    

DOI:10.1089/hum.1993.4.1-25

出版商:Mary Ann Liebert, Inc.    

年代:1993

数据来源: MAL

ISSN:1043-0342    

DOI:10.1089/hum.1993.4.1-35

出版商:Mary Ann Liebert, Inc.    

年代:1993

数据来源: MAL

摘要:

SCIENTIFIC ABSTRACTMurine retroviral vectors can infect a wide variety of proliferating mammalian cell types (e.g. lymphocytes). Non-proliferating tissues (e.g. neurons) are not transduced by murine retroviral vectors. These findings suggest that this type of vector may be useful for the selective introduction of genes into growing tumors in the brain, since the tumor is essentially the only tissue that would integrate and express the vector genes.We investigated the possibility ofin vivotransduction of growing brain tumors with the herpes simplex thymidine kinase (HS-tk) gene. Rats with a malignant brain tumor were given an intratumoral stereotaxic injection of murine fibroblasts that were producing a retroviral vector containing the herpes simplex thymidine kinase (HS-tk) gene or a control vector producer cell line containing the-galactosidase gene. The animals were rested 5 days to allow time for the HS-tk retroviral vectors that were producedin situto transduce the neighboring proliferating glioma. The animals were then treated with the anti-herpes drug ganciclovir (GCV). Gliomas injected with the HS-tk producer cells regressed completely with GCV therapy while the tumors injected with-galactosidase producers had large tumors. Staining for-galactosidase positive cells in control animal brain revealed transduction of 4080% of the tumor cells without evidence of transduction of the surrounding normal brain tissue. No significant toxicity was observed in toxicity studies in mice, rats and non-human primates.Based upon these findings, we have proposed a human clinical trial to determine if the direct injection of the G1TkSvNa producer cell line into growing human brain tumors will regress with GCV therapy. The patient population consists of individuals with recurrent malignant tumors who have failed standard therapy for their primary or metastatic brain tumors. The expected survival of these patients is limited to weeks to a few months.

ISSN:1043-0342    

DOI:10.1089/hum.1993.4.1-39

出版商:Mary Ann Liebert, Inc.    

年代:1993

数据来源: MAL

ISSN:1043-0342    

DOI:10.1089/hum.1993.4.1-71

出版商:Mary Ann Liebert, Inc.    

年代:1993

数据来源: MAL

ISSN:1043-0342    

DOI:10.1089/hum.1993.4.1-87

出版商:Mary Ann Liebert, Inc.    

年代:1993

数据来源: MAL

ISSN:1043-0342    

DOI:10.1089/hum.1993.4.1-91

出版商:Mary Ann Liebert, Inc.    

年代:1993

数据来源: MAL