A quick and efficient method to generate mammalian stable cell lines based on a novel inducible alphavirus DNA/RNA layered system

Aranda, Alejandro; Bezunartea, Jaione; Casales, Erkuden; Rodríguez-Madoz, Juan R.; Larrea, Esther; Prìeto, Jesús; Smerdou, Cristian

doi:10.1007/s00018-014-1631-2

Cited by 6 publications

(4 citation statements)

References 36 publications

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“…The coding sequence of hOSM was obtained as previously described [ 45 ]. The genome of Ad-OSM was based on the previously described virus Ad-IL12G [ 32 ], in which the single chain IL-12 coding sequence was replaced by hOSM, using standard molecular biology techniques.…”

Section: Methodsmentioning

confidence: 99%

Enhanced therapeutic effect using sequential administration of antigenically distinct oncolytic viruses expressing oncostatin M in a Syrian hamster orthotopic pancreatic cancer model

et al. 2015

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BackgroundThe limited efficacy of current treatments against pancreatic cancer has prompted the search of new alternatives such as virotherapy. Activation of the immune response against cancer cells is emerging as one of the main mechanisms of action of oncolytic viruses (OV). Direct oncolysis releases tumor antigens, and viral replication within the tumor microenvironment is a potent danger signal. Arming OV with immunostimulatory transgenes further enhances their therapeutic effect. However, standard virotherapy protocols do not take full advantage of OV as cancer vaccines because repeated viral administrations may polarize immune responses against strong viral antigens, and the rapid onset of neutralizing antibodies limits the efficacy of redosing. An alternative paradigm based on sequential combination of antigenically distinct OV has been recently proposed.MethodsWe have developed a protocol consisting of sequential intratumor administrations of new Adenovirus (Ad) and Newcastle Disease Virus (NDV)-based OV encoding the immunostimulatory cytokine oncostatin M (OSM). Transgene expression, toxicity and antitumor effect were evaluated using an aggressive orthotopic pancreatic cancer model in Syrian hamsters, which are sensitive to OSM and permissive for replication of both OVs.ResultsNDV-OSM was more cytolytic, whereas Ad-OSM caused higher OSM expression in vivo. Both viruses achieved only a marginal antitumor effect in monotherapy. In addition, strong secretion of OSM in serum limited the maximal tolerated dose of Ad-OSM. In contrast, moderate doses of Ad-OSM followed one week later by NDV-OSM were safe, showed a significant antitumor effect and stimulated immune responses against cancer cells. Similar efficacy was observed when the order of virus administrations was reversed.ConclusionSequential administration of oncolytic Ad and NDV encoding OSM is a promising approach against pancreatic cancer.Electronic supplementary materialThe online version of this article (doi:10.1186/s12943-015-0479-x) contains supplementary material, which is available to authorized users.

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Section: Methodsmentioning

confidence: 99%

Enhanced therapeutic effect using sequential administration of antigenically distinct oncolytic viruses expressing oncostatin M in a Syrian hamster orthotopic pancreatic cancer model

et al. 2015

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“…Furthermore, this change allows the expression of a highly pure and bioactive protein, potentially reducing the production costs, since it is possible to produce a greater amount of hGDNF using a smaller number of electroporations and a lower amount of RNA. Even though the quantity of hGDNF produced at a small scale in this study was sufficient for our objective, it would be very interesting to scale-up hGDNF production using this expression system (Núñez et al, 2013, Lundstrom, 2003, Aranda et al, 2014. The ideal situation would be to adapt this method to a process able to handle large volumes of cells with suitable bioreactors.…”

Section: Future Prospects and Conclusionmentioning

confidence: 99%

“…In recent years, many studies have focused on the development of vectors with a less cytopathic character. However, the complexity of controlling the duration of the protein expression remains a major limitation (Aranda et al, 2014, Casales et al, 2010, Casales et al, 2008, Schott et al, 2016. On the other hand, cell electroporation is a discontinuous procedure, which could represent a limitation for scaling-up our method.…”

Section: Future Prospects and Conclusionmentioning

confidence: 99%

Optimization of a GDNF production method based on Semliki Forest virus vector

Torres-Ortega

Smerdou

Ansorena

et al. 2021

European Journal of Pharmaceutical Sciences

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“…Recombinant proteins have been expressed transiently, [162][163][164] constitutively 165 or inducibly. 166 Inducible cell lines contain an integrated version of the cDNA replicon sequence and offer the advantage of producing even toxic proteins, such as oncostatin M, at high level as cell clones can be expanded to the desired amount prior to induction. 166 Successful production has been reported for recombinant proteins as diverse as prokaryotic (e.g., bacterial β-galactosidase), 163 viral (HIV-1 gp160 and gp120) 162 and eukaryotic proteins for human therapeutic use, including 5-HT(3) serotonin receptor, 164 insulin-like growth factor I, and cardiotrophin-1.…”

Section: Transmission-competent Particlesmentioning

confidence: 99%

Viral and Synthetic RNA Vector Technologies and Applications

et al. 2016

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Use of RNA is an increasingly popular method to transiently deliver genetic information for cell manipulation in basic research and clinical therapy. In these settings, viral and nonviral RNA platforms are employed for delivery of small interfering RNA and protein-coding mRNA. Technological advances allowing RNA modification for increased stability, improved translation and reduced immunogenicity have led to increased use of nonviral synthetic RNA, which is delivered in naked form or upon formulation. Alternatively, highly efficient viral entry pathways are exploited to transfer genes of interest as RNA incorporated into viral particles. Current viral RNA transfer technologies are derived from Retroviruses, nonsegmented negative-strand RNA viruses or positive-stranded Alpha- and Flaviviruses. In retroviral particles, the genes of interest can either be incorporated directly into the viral RNA genome or as nonviral RNA. Nonsegmented negative-strand virus-, Alpha- and Flavivirus-derived vectors support prolonged expression windows through replication of viral RNA encoding genes of interest. Mixed technologies combining viral and nonviral components are also available. RNA transfer is ideal for all settings that do not require permanent transgene expression and excludes potentially detrimental DNA integration into the target cell genome. Thus, RNA-based technologies are successfully applied for reprogramming, transdifferentiation, gene editing, vaccination, tumor therapy, and gene therapy.

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A quick and efficient method to generate mammalian stable cell lines based on a novel inducible alphavirus DNA/RNA layered system

Cited by 6 publications

References 36 publications

Enhanced therapeutic effect using sequential administration of antigenically distinct oncolytic viruses expressing oncostatin M in a Syrian hamster orthotopic pancreatic cancer model

Enhanced therapeutic effect using sequential administration of antigenically distinct oncolytic viruses expressing oncostatin M in a Syrian hamster orthotopic pancreatic cancer model

Optimization of a GDNF production method based on Semliki Forest virus vector

Viral and Synthetic RNA Vector Technologies and Applications

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