Specific Regression of Human Cancer Cells by Ribozyme-Mediated Targeted Replacement of Tumor-Specific Transcript

Kwon, Byung Su; Jung, Heung Su; Song, Min Jong; Cho, Kyung Sook; Kim, Sung Chun; Kimm, Kuchan; Jeong, Jin Sook; Kim, In Hoo; Lee, Seong Wook

doi:10.1016/j.ymthe.2005.06.096

Cited by 47 publications

(70 citation statements)

References 30 publications

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“…In contrast, no accessible sites were found in the long hairpin-formed CUG repeat region. In previous studies, we showed that the relative trans-splicing reaction efficacy at different sites in a target RNA in cells and in vitro was closely associated with the predicted accessibility determined by mapping results (Ryu et al, 2003;Ryu and Lee, 2004;Kwon et al, 2005). Therefore, the ribozymes that target the ac-cessible sites identified here are anticipated to be more active than those that target other sites.…”

Section: Ribozyme-accessible Uridines In Mdmpk 3'-utr Rnamentioning

confidence: 53%

“…To this end, we employed an RNA mapping strategy based on RNA tagging (Jones et al,1996) and a trans-splicing ribozyme library, which has been described in detail elsewhere (Fig. 1) (Lan et al, 1998;Lan et al, 2000;Kwon et al, 2005;Ryu et al, 2003;Song and Lee, 2007).…”

Section: Mapping Of the Trans-splicing Ribozymementioning

confidence: 99%

“…1998; Phylactou et al 1998;Rogers et al, 2002). Moreover, we have shown that trans-splicing ribozyme targeting and replacing the human telomerase reverse transcriptase (hTERT) RNA could specifically and efficiently regress human tumors not only in cell cultures but also in animals without any toxicity in normal tissues or any off-target splicing effects Kwon et al, 2005;Song and Lee, 2008;Song et al, 2009). Thus, a trans-splicing ribozyme that can repair toxic mDMPK RNA with the wild-type one or replace the nucleus-entrapped mutant transcripts with RNA that can be released into the cytoplasm would be a potential tool for DM1 treatment.…”

Section: Introductionmentioning

confidence: 99%

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RNA Mapping of Mutant Myotonic Dystrophy Protein Kinase 3'-Untranslated Region Transcripts

Song¹,

Lee²

2009

Genomics & Informatics

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Myotonic dystrophy type 1 (DM1), which is a dominantly inherited neurodegenerative disorder, results from a CTG trinucleotide repeat expansion in the 3'-untranslated region (3'-UTR) of the myotonic dystrophy protein kinase (DMPK) gene. Retention of mutant DMPK (mDMPK) transcripts in the nuclei of affected cells has been known to be the main cause of pathogenesis of the disease. Thus, reducing the RNA toxicity through elimination of the mutant RNA has been suggested as one therapeutic strategy against DM1. In this study, we suggested RNA replacement with a trans-splicing ribozyme as an alternate genetic therapeutic approach for amelioration of DM1. To this end, we identified the regions of mDMPK 3'-UTR RNA that were accessible to ribozymes by using an RNA mapping strategy based on a transsplicing ribozyme library. We found that particularly accessible sites were present not only upstream but also downstream of the expanded repeat sequence. Repair or replacement of the mDMPK transcript with the specific ribozyme will be useful for DM1 treatment through reduction of toxic mutant transcripts and simultaneously restore wild-type DMPK or release nucleus-entrapped mDMPK transcripts to the cytoplasm.

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Section: Ribozyme-accessible Uridines In Mdmpk 3'-utr Rnamentioning

confidence: 53%

Section: Mapping Of the Trans-splicing Ribozymementioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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RNA Mapping of Mutant Myotonic Dystrophy Protein Kinase 3'-Untranslated Region Transcripts

Song¹,

Lee²

2009

Genomics & Informatics

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“…11,19 In brief, the Rib21AS ribozyme targeting U21 on the hTERT RNA was generated to harbor the extended internal guide sequence, which includes as an extension of the P1 helix, an additional 6-nt-long P10 helix, and a 325-nt-long antisense sequence complementary to the downstream region of the targeted hTERT RNA uridine. 11 cDNA as a 3 0 exon encoding for lacZ or herpes simplex virus thymidine kinase (HSVtk) genes was inserted into NruI/XbaI downstream of the modified group I intron expression construct. The resulting ribozyme was then cloned into pcDNA or pPEPCK-LCR (kindly provided by Dr K Oka from the Bayler College of Medicine).…”

Section: Generation Of Recombinant Adenoviral Vectorsmentioning

confidence: 99%

“…We validated this concept by developing transsplicing ribozymes targeting and replacing human telomerase reverse transcriptase (hTERT) RNA to induce transgene activity specifically in cancer cells that express the RNA, demonstrating that the ribozyme can specifically mark tumor cells expressing hTERT or specifically make tumor cells sensitive to prodrug treatment not only in vitro but in tumor xenografts. [11][12][13] This study showed that a trans-splicing ribozyme provides a multifunctional gene therapy tool that both targets and treats the tumor in two ways. A major advantage of this approach is that by invading a tumor-specific pre-mRNA, targeted expression of the therapeutic gene product is guaranteed, with a simultaneous reduction in the expression of the target gene product resulting in an additive, or perhaps a synergistic, anticancer effect.…”

Section: Introductionmentioning

confidence: 99%

Validation of tissue-specific promoter-driven tumor-targeting trans-splicing ribozyme system as a multifunctional cancer gene therapy device in vivo

Ban

et al. 2008

Cancer Gene Ther

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A trans-splicing ribozyme that can specifically reprogram human telomerase reverse transcriptase (hTERT) RNA was previously suggested as a useful tool for tumor-targeted gene therapy. In this study, we applied transcriptional targeting with the RNA replacement approach to target liver cancer cells by combining a liver-selective promoter with an hTERT-mediated cancer-specific ribozyme. To validate effects of this system in vivo, we constructed an adenovirus encoding for the hTERT-targeting trans-splicing ribozyme under the control of a liver-selective phosphoenolpyruvate carboxykinase promoter. We observed that intratumoral injection of this virus produced selective and efficient regression of tumors that had been subcutaneously inoculated with hTERTpositive liver cancer cells in mice. Importantly, the trans-splicing reaction worked equally well in a nude mouse model of hepatocarcinoma-derived peritoneal carcinomatosis, inducing the highly specific expression of a transgene, and moreover, the efficient regression of the hTERT-positive liver tumors with minimal liver toxicity when systemically delivered with the adenovirus. In addition to the observed hTERT-dependent therapeutic gene induction, significant reductions in the levels of hTERT RNA (B75%) were also observed. In conclusion, this study demonstrates that a cancer-specific RNA replacement approach using transsplicing ribozyme with a tissue-selective promoter represents a promising strategy for cancer treatment.

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Imaging Target mRNA and siRNA‐Mediated Gene Silencing In Vivo with Ribozyme‐Based Reporters

Gowrishankar

Hasegawa

et al. 2008

ChemBioChem

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Noninvasive imaging of specific mRNAs in living subjects promises numerous biological and medical applications. Common strategies use fluorescently or radioactively labelled antisense probes to detect target mRNAs through a hybridization mechanism, but have met with limited success in living animals. Here we present a novel molecular imaging approach based on the group I intron of Tetrahymena thermophila for imaging mRNA molecules in vivo. Engineered trans-splicing ribozyme reporters contain three domains, each of which is designed for targeting, splicing, and reporting. They can transduce the target mRNA into a reporter mRNA, leading to the production of reporter enzymes that can be noninvasively imaged in vivo. We have demonstrated this ribozyme-mediated RNA imaging method for imaging a mutant p53 mRNA both in single cells and noninvasively in living mice. After optimization, the ribozyme reporter increases contrast for the transiently expressed target by 180-fold, and by ten-fold for the stably expressed target. siRNA-mediated specific gene silencing of p53 expression has been successfully imaged in real time in vivo. This new ribozyme-based RNA reporter system should open up new avenues for in vivo RNA imaging and direct imaging of siRNA inhibition.

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Specific Regression of Human Cancer Cells by Ribozyme-Mediated Targeted Replacement of Tumor-Specific Transcript

Cited by 47 publications

References 30 publications

RNA Mapping of Mutant Myotonic Dystrophy Protein Kinase 3'-Untranslated Region Transcripts

RNA Mapping of Mutant Myotonic Dystrophy Protein Kinase 3'-Untranslated Region Transcripts

Validation of tissue-specific promoter-driven tumor-targeting trans-splicing ribozyme system as a multifunctional cancer gene therapy device in vivo

Imaging Target mRNA and siRNA‐Mediated Gene Silencing In Vivo with Ribozyme‐Based Reporters

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