Application of Targeted Radiotherapy/Gene Therapy to Bladder Cancer Cell Lines

Fullerton, Natasha E; Mairs, Robert J.; Kirk, D.; Keith, W. Nicol; Carruthers, Ross; McCluskey, A.; Brown, Mary; Wilson, Lesley; Boyd, Marie

doi:10.1016/j.eururo.2004.09.009

Cited by 13 publications

(13 citation statements)

References 42 publications

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“…Generally mammalian promoters are characterized by low-expression activity compared with viral regulatory elements, resulting in inefficient activation of transcription and, consequently, low levels of production of therapeutic protein (36). However, we demonstrated previously that the hTR and hTERT promoters were comparable to the strong ubiquitous RSV and CMV promoters with respect to transgene expression in vitro (3,4,20,21).…”

Section: Discussionmentioning

confidence: 99%

“…Recombinant plasmids containing the bNAT cDNA under control of the CMV, hTR, or hTERT promoter were constructed as previously described (2,3,21).…”

Section: Plasmidsmentioning

confidence: 99%

“…The tissue was homogenized and RNA was extracted according to the manufacturer's instructions. Preparation of cDNA and RT-PCR were performed using TaqMan RT-PCR methodology and reagents (Perkin-Elmer Applied Biosystems), as described previously (21,27). Primer and probe sequences were designed from published sequences for bNAT (accession no.…”

Section: Real-time Reverse Transcriptase Polymerase Chain Reaction (Rmentioning

confidence: 99%

“…UVW or EJ138 parental cell lines were stably transfected with the recombinant plasmids using Effectene transfection reagent (Qiagen) as previously described (2)(3)(4)(5)21). The transfectants were maintained by addition of geneticin (G418) at every passage, and stable transfectants were not further selected into clones derived from single transfected cells.…”

Section: Cell Culture and Transfection Of Nat Genementioning

confidence: 99%

“…Subsequently, we refined this therapeutic scheme by using the tumor-specific telomerase promoters (hTR and hTERT) to control NAT expression (3). Unlike most mammalian control elements, these were strong inducers of expression in vitro, similar in potency to that of the ubiquitous CMV promoter (4,20,21).…”

mentioning

confidence: 99%

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A Transfectant Mosaic Xenograft Model for Evaluation of Targeted Radiotherapy in Combination with Gene Therapy In Vivo

Mairs¹,

Ross

McCluskey

et al. 2007

Journal of Nuclear Medicine

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For gene therapy to be efficacious in the treatment of cancer, therapeutic transgenes must be limited in their expression to tumor cells and must be expressed at sufficiently high transcriptional levels. Moreover, the inadequacy of gene delivery must be overcome by induction of toxicity to neighboring nontargeted cells. Combining targeted radionuclide therapy with gene therapy using human telomerase promoters has shown promise in these respects, and the efficacy of this scheme has been assessed in vitro using transfectant mosaic tumor spheroids. To enable the evaluation of targeted radiotherapy combined with gene transfer in vivo, we have developed a transfectant mosaic xenograft (TMX) model. Methods: Human telomerase promoters were used to drive expression of the noradrenaline transporter (NAT) transgene in 2 human cell lines (UVW and EJ138). Promoter activity was assessed in xenografts in nude mice by determination of the uptake of the radiopharmaceutical 131 I-metaiodobenzylguanidine ( 131 I-MIBG) and by measurement of tumor growth. The efficacy of 131 I-MIBG treatment was also assessed in TMXs to determine the delay in growth of tumors composed of various proportions of NAT-expressing cells-a likely clinical scenario after gene delivery in vivo. Results: In terms of induction of the capacity for active uptake of 131 I-MIBG and the resultant inhibition of tumor growth in vivo, both telomerase promoters (hTR and hTERT) were similar in potency to the CMV (cytomegalovirus) promoter as controlling elements for the expression of the NAT transgene. In TMXs derived from UVW and EJ138 cells, 131 I-MIBG uptake was proportional to NAT gene expression (r s 5 0.910, P , 0.001 for UVW; r s 5 0.971, P , 0.001 for EJ138). Inhibition of the growth of these tumors correlated with the fraction of NATtransfected cells (r s 5 0.910, P , 0.001 for UVW; r s 5 0.971, P , 0.001 for EJ138), and substantial tumor growth delay was observed when 5% of the xenograft was composed of NATpositive cells. Conclusion: TMXs constitute a suitable model to measure the efficacy of cancer gene therapy strategies when ,100% of the tumor mass can be targeted to express the therapeutic transgene.

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Section: Discussionmentioning

confidence: 99%

“…Recombinant plasmids containing the bNAT cDNA under control of the CMV, hTR, or hTERT promoter were constructed as previously described (2,3,21).…”

Section: Plasmidsmentioning

confidence: 99%

Section: Real-time Reverse Transcriptase Polymerase Chain Reaction (Rmentioning

confidence: 99%

Section: Cell Culture and Transfection Of Nat Genementioning

confidence: 99%

mentioning

confidence: 99%

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A Transfectant Mosaic Xenograft Model for Evaluation of Targeted Radiotherapy in Combination with Gene Therapy In Vivo

Mairs¹,

Ross

McCluskey

et al. 2007

Journal of Nuclear Medicine

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The Norepinephrine Transporter in Physiology and Disease

Bönisch

Brüss

Handbook of Experimental Pharmacology

108

101

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Three-dimensional neuroblastoma cell culture: proteomic analysis between monolayer and multicellular tumor spheroids

et al. 2008

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Introduction-Solid tumors, such as neuroblastoma (NB), are associated with a heterogeneous cell environment. Multicellular tumor spheroid (MCTS) cultures have been shown to better mimic growth characteristics of in vivo solid tumors. Because tumor spheroid growth patterns may be quite different from standard two-dimensional culture systems, we sought to compare the protein expression profiles of two-and three-dimensional in vitro NB cultures, i.e., monolayers and MCTS.

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Application of Targeted Radiotherapy/Gene Therapy to Bladder Cancer Cell Lines

Cited by 13 publications

References 42 publications

A Transfectant Mosaic Xenograft Model for Evaluation of Targeted Radiotherapy in Combination with Gene Therapy In Vivo

A Transfectant Mosaic Xenograft Model for Evaluation of Targeted Radiotherapy in Combination with Gene Therapy In Vivo

The Norepinephrine Transporter in Physiology and Disease

Three-dimensional neuroblastoma cell culture: proteomic analysis between monolayer and multicellular tumor spheroids

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