Mutagenesis and Oncogenesis by Chromosomal Insertion of Gene Transfer Vectors

Baum, Christopher F.; Kustikova, Olga; Modlich, Ute; Li, Zhixiong

doi:10.1089/hum.2006.17.ft-181

Cited by 7 publications

(9 citation statements)

References 97 publications

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“…This creates the risk of insertional mutagenesis, which can be caused by transactivation of proto-oncogenes through semi-randomly integrated vectorderived enhancer/promoter sequences. 41 The associated risk can be circumvented in iPSC-based therapies because the potential for unlimited proliferation allows in vitro selection of clonal iPSC lines with defined integration sites and without genetic alterations. In an extensive study, including lentiviral reprogramming of patient cells, Cre-mediated vector excision, and secondary lentiviral gene transfer, Papapetrou et al showed that such a selection of iPSC clones with single vector integrations of a lentiviral β-globin expression vector is feasible.…”

Section: Discussionmentioning

confidence: 99%

Modified Lentiviral LTRs Allow Flp Recombinase–mediated Cassette Exchange and In Vivo Tracing of “Factor-free” Induced Pluripotent Stem Cells

et al. 2014

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Methods for generating induced pluripotent stem cells (iPSCs) for disease modeling and cell therapies have progressed from integrating vectors to transient delivery of reprogramming factors, avoiding permanent genomic modification. A major limitation of unmodified iPSCs is the assessment of their distribution and contribution to adverse reactions in autologous cell therapy. Here, we report that polycistronic lentiviral vectors with single Flp recombinase (Flp) recognition target (FRT) sites can be used to generate murine iPSCs that are devoid of the reprogramming cassette but carry an intergenic 300-bp long terminal repeat sequence. Performing quantitative polymerase chain reaction on this marker, we could determine genetic identity and tissue contribution of iPSC-derived teratomas in mice. Moreover, we generated iPSCs carrying heterospecific FRT twin sites, enabling excision and recombinase-mediated cassette exchange (RMCE) of the reprogramming cassette for another expression unit of choice. Following screening of iPSCs for "safe harbor" integration sites, expression cassettes were introduced by RMCE into various previously silenced loci of selected single-copy iPSCs. Analysis of DNA methylation showed that RMCE reverted the local epigenetic signature, which allowed transgene expression in undifferentiated iPSCs and in differentiated progeny. These findings support the concept of creating clonotypically defined exchangeable and traceable pluripotent stem cells for disease research and cell therapy.

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Section: Discussionmentioning

confidence: 99%

Modified Lentiviral LTRs Allow Flp Recombinase–mediated Cassette Exchange and In Vivo Tracing of “Factor-free” Induced Pluripotent Stem Cells

et al. 2014

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“…Viruses such as lentiviruses, adenoviruses, and adeno-associated viruses can efficiently deliver vectors encoding miRNA mimics and anti-miRNAs into nuclei (63)(64)(65). However, critical hurdles such as insertional mutagenesis, immunogenicity, and limited packaging capacity arise when using these viral vectors (66,67 Therapeutic manipulation of the ATM-miRNA axis Given recent advances in our knowledge of the crucial roles of ATM-regulated miRNAs and their dysregulation in cancers, modulation of miRNA expression provides a promising therapeutic strategy for cancer (71). In the following sections, we discuss the therapeutic modulation of ATM-regulated miRNAs, including reintroduction of tumor-suppressive miRNA, oncomiR inhibition, and some technical considerations regarding their delivery systems and tissue specificity.…”

Section: In Vivo Mirna Delivery Strategiesmentioning

confidence: 99%

Therapeutic Potential of the miRNA–ATM Axis in the Management of Tumor Radioresistance

2020

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The ataxia-telangiectasia mutated (ATM) protein kinase is widely known for its function as a chief mobilizer of the DNA damage response (DDR) upon DNA double-strand breaks. ATM orchestrates the DDR by modulating the expression of various miRNAs through several mechanisms. On the other hand, a set of miRNAs contribute to tight regulation of ATM by directly targeting the 3 0 -untranslated region of ATM mRNA. This review addresses the therapeutic application and molecular mechanisms that underlie the intricate interactions between miRNAs and ATM. It also describes therapeutic delivery of miRNAs in different environments such as hypoxic tumor microenvironments.

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“…6–8 Successful gene delivery examples have been extensively achieved based on the viral vectors due to their excellent transfection efficiencies. 9–11 However, the viral vectors often suffer from severe immunogenicity, insertional mutagenesis and oncogenicity which were associated with serious safety concerns. 10,12,13 Recently, synthetic non-viral vectors, mainly exemplified by cationic polymers (polycations) and cationic lipids, have attracted great attentions as safer and more promising alternatives to conventional viral vectors owing to their low immunogenicity and oncogenicity.…”

Section: Introductionmentioning

confidence: 99%

“…9–11 However, the viral vectors often suffer from severe immunogenicity, insertional mutagenesis and oncogenicity which were associated with serious safety concerns. 10,12,13 Recently, synthetic non-viral vectors, mainly exemplified by cationic polymers (polycations) and cationic lipids, have attracted great attentions as safer and more promising alternatives to conventional viral vectors owing to their low immunogenicity and oncogenicity. 14–18 However, the clinical application of non-viral vectors is often limited by the relatively lower transfection efficiencies compared to viral vectors due to various extra- and intracellular barriers.…”

Section: Introductionmentioning

confidence: 99%

Thioketal-crosslinked: ROS-degradable polycations for enhanced in vitro and in vivo gene delivery with self-diminished cytotoxicity

et al. 2019

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The irreversible correlation between the transfection efficiency and cytotoxicity of polycations has always been the huge obstacle that severely limited the gene delivery efficiency. The undesired inconsistency could be mainly attributed to the molecular weight (MW) of the polycations, that is, polymers with high MW and positive charge densities exhibited enhanced transfection efficiency but with associated cytotoxicity. To address such critical challenge, we developed the degradable high MW polymers strategy which could condense DNA for the internalization and release DNA upon the trigger-responsive degradation. In this work, two kinds of degradable PEI-based polymers were prepared via the polyaddition of PEI600 and dienes with ester groups or thiol ketal groups. The PEI-based degradable polymers could efficiently condense plasmid DNA to form the nano-complexes with the size around 180 nm. They also exhibited improved gene delivery efficiency compared with commercial available transfection reagent, PEI25k, and were 380–520 folds higher than PEI600 in HeLa cells. The toxicity of the polymers could be reduced by the rapid degradation upon acid or ROS triggering as well as intracellular release of DNA and the cell viability could reach higher than 80% even at high doses. ROS-responsive PEI-based polymers also demonstrated its potential applications especially in cancer cells, which were proved by the enhanced in vivo gene expression in cancer cells. Our strategy therefore allows an effective tool to manipulate the relationship between transfection efficiency and cytotoxicity of polycations, and thus provides an effective insight into the rational design of non-viral gene delivery vectors.

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Mutagenesis and Oncogenesis by Chromosomal Insertion of Gene Transfer Vectors

Cited by 7 publications

References 97 publications

Modified Lentiviral LTRs Allow Flp Recombinase–mediated Cassette Exchange and In Vivo Tracing of “Factor-free” Induced Pluripotent Stem Cells

Modified Lentiviral LTRs Allow Flp Recombinase–mediated Cassette Exchange and In Vivo Tracing of “Factor-free” Induced Pluripotent Stem Cells

Therapeutic Potential of the miRNA–ATM Axis in the Management of Tumor Radioresistance

Thioketal-crosslinked: ROS-degradable polycations for enhanced in vitro and in vivo gene delivery with self-diminished cytotoxicity

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