Potential Therapeutic Applications of Rna Cap Analogs

Ziemniak, Marcin; Strenkowska, Malwina; Kowalska, Joanna; Jemielity, Jacek

doi:10.4155/fmc.13.96

Cited by 61 publications

(70 citation statements)

References 165 publications

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“…The 5'-cap of mRNA has been reported to be associated with mRNA transcription and translation, preventing the degradation of mRNA precursors, and serving as a binding site for regulatory factors, therefore, being involved in various RNA metabolic processes (3). By binding to the 5'-cap of mRNA, eIF4E forms a translation initiation complex involved in protein trafficking and translation, thereby regulating gene expression (4,5).…”

Section: Introductionmentioning

confidence: 99%

Knockdown of eukaryotic translation initiation factor 4E suppresses cell growth and invasion, and induces apoptosis and cell cycle arrest in a human lung adenocarcinoma cell line

Chen

Zhang

Xia

et al. 2016

Molecular Medicine Reports

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Abstract. Eukaryotic translation initiation factor 4E (eIF4E) was shown to be upregulated in malignant human tumors. To assess the effect of downregulation of eIF4E on the proliferation and invasiveness of a human lung adenocarcinoma cell line, a short hairpin (sh)RNA targeting eIF4E was constructed and transfected into A549 human lung adenocarcinoma cells. The expression of eIF4E was determined by reverse transcription-quantitative polymerase chain reaction and western blotting. Cell viability was assessed using a Cell Counting kit-8, and apoptosis levels and cell cycle distribution were assessed by flow cytometry. Invasiveness was assessed using Transwell chambers. Transfection of the A549 cells with eIF4E targeting shRNA reduced the mRNA and protein expression levels of eIF4E by >70% 48 and 72 h following transfection, and eIF4E targeting shRNA-transfected cells were significantly less viable compared with the cells transfected with scrambled shRNA. The rate of apoptosis was also significantly increased, significantly more cells were in the G 0 /G 1 phase and fewer were in the S phase, indicating cell cycle arrest. The fraction of transfected cells migrating across Transwell inserts were also reduced. In conclusion, inhibition of eIF4E suppressed cell growth and invasion, induced apoptosis and cell cycle arrest, suggesting that eIF4E may be a potential therapeutic target in lung adenocarcinoma.

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Section: Introductionmentioning

confidence: 99%

Knockdown of eukaryotic translation initiation factor 4E suppresses cell growth and invasion, and induces apoptosis and cell cycle arrest in a human lung adenocarcinoma cell line

Chen

Zhang

Xia

et al. 2016

Molecular Medicine Reports

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“…The cap plays a crucial role in several cellular processes, such as pre-mRNA splicing (Sharp 1994), transport of RNA from the nucleus to the cytoplasm (Lewis and Izaurralde 1997), protection of mRNA against degradation (Rhoads 1985), and translation initiation (Gingras et al 1999). Over the years, chemically modified cap analogs have been used for biochemical, biophysical, and structural studies on mechanisms of the aforementioned processes (Niedzwiecka et al 2002;Grudzien et al 2004;Jankowska-Anyszka et al 2011;Ziemniak et al 2013). So far, some of the most intense research directed toward the application of synthetic cap analogs has been focused on designing translation inhibitors that prevent binding of mRNA to eIF4E.…”

Section: All Cellular Eukaryotic Mrnas Have At Theirmentioning

confidence: 99%

“…It was shown that elements such as (1) the positive charge of the imidazole ring of guanosine, (2) the presence of at least two negative charges (or a negative charge and the free electron pair) within the phosphate chain, and (3) the presence of at least one hydrogen atom on the amino group of the 7-methyloguanosine should be preserved in order to synthesize an efficient translation inhibitor. To date, numerous modified cap analogs have been tested as inhibitors of cap-dependent translation in vitro by competing with mRNA for the binding site of eIF4E (Cai et al 1999;Grudzien et al 2004;Ghosh et al 2005;Jia et al 2010;Piecyk et al 2012;Ziemniak et al 2013). Unfortunately, due to their charged nature, in vivo application of such analogs is strictly limited.…”

Section: All Cellular Eukaryotic Mrnas Have At Theirmentioning

confidence: 99%

Triazole-containing monophosphate mRNA cap analogs as effective translation inhibitors

Piecyk

Łukaszewicz

Darżynkiewicz

et al. 2014

RNA

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Synthetic analogs of the 5′ end of mRNA (cap structure) are widely used in molecular studies on mechanisms of cellular processes such as translation, intracellular transport, splicing, and turnover. The best-characterized cap binding protein is translation initiation factor 4E (eIF4E). Recognition of the mRNA cap by eIF4E is a critical, rate-limiting step for efficient translation initiation and is considered a major target for anticancer therapy. Here, we report a facile methodology for the preparation of N2-triazole-containing monophosphate cap analogs and present their biological evaluation as inhibitors of protein synthesis. Five analogs possessing this unique hetero-cyclic ring spaced from the m7-guanine of the cap structure at a distance of one or three carbon atoms and/or additionally substituted by various groups containing the benzene ring were synthesized. All obtained compounds turned out to be effective translation inhibitors with IC 50 similar to dinucleotide triphosphate m 7 GpppG. As these compounds possess a reduced number of phosphate groups and, thereby, a negative charge, which may support their cell penetration, this type of cap analog might be promising in terms of designing new potential therapeutic molecules. In addition, an exemplary dinucleotide from a corresponding mononucleotide containing benzyl substituted 1,2,3-triazole was prepared and examined. The superior inhibitory properties of this analog (10-fold vs. m 7 GpppG) suggest the usefulness of such compounds for the preparation of mRNA transcripts with high translational activity.

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“…A major obstacle to understanding cap recognition by Dcp2 is that all previously studied m 7 GpppN cap analogs bind the enzyme with very weak (millimolar) affinities. We have previously identified several phosphate-modified cap analogs which, after incorporation at the 5 ′ terminus of mRNAs, make them more resistant to Dcp2-mediated decapping (GrudzienNogalska et al 2013;Ziemniak et al 2013a). The modifications include bridging substitution at the α/β-position of the cap (O-to-CH 2 or methylenebisphosphonate; O-to-NH or imidodiphosphate) (Grudzien et al 2006;Su et al 2011) or nonbridging substitution at the β-phosphate (O-to-S or phosphorothioate; O-to-BH 3 or boranophosphate) (Supplemental Fig.…”

Section: Introductionmentioning

confidence: 99%

“…Some of the analogs that made mRNA resistant to Dcp2 also bound the translational cap-binding protein eIF4E with higher affinity and, consequently, produced mRNA with both increased cellular half-life and increased translational efficiency. These cap-modified transcripts have found application in experimental mRNA-based therapies (Kuhn et al 2010;Ziemniak et al 2013a;Sahin et al 2014). …”

Section: Introductionmentioning

confidence: 99%

Two-headed tetraphosphate cap analogs are inhibitors of the Dcp1/2 RNA decapping complex

Ziemniak

Mugridge

Kowalska

et al. 2016

RNA

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Dcp1/2 is the major eukaryotic RNA decapping complex, comprised of the enzyme Dcp2 and activator Dcp1, which removes the 5 ′ m 7 G cap from mRNA, committing the transcript to degradation. Dcp1/2 activity is crucial for RNA quality control and turnover, and deregulation of these processes may lead to disease development. The molecular details of Dcp1/2 catalysis remain elusive, in part because both cap substrate (m 7 GpppN) and m 7 GDP product are bound by Dcp1/2 with weak (mM) affinity. In order to find inhibitors to use in elucidating the catalytic mechanism of Dcp2, we screened a small library of synthetic m 7 G nucleotides (cap analogs) bearing modifications in the oligophosphate chain. One of the most potent cap analogs, m 7 Gp S ppp S m 7 G, inhibited Dcp1/2 20 times more efficiently than m 7 GpppN or m 7 GDP. NMR experiments revealed that the compound interacts with specific surfaces of both regulatory and catalytic domains of Dcp2 with submillimolar affinities. Kinetics analysis revealed that m 7 Gp S ppp S m 7 G is a mixed inhibitor that competes for the Dcp2 active site with micromolar affinity. m 7 Gp S ppp S m 7 G-capped RNA undergoes rapid decapping, suggesting that the compound may act as a tightly bound cap mimic. Our identification of the first small molecule inhibitor of Dcp2 should be instrumental in future studies aimed at understanding the structural basis of RNA decapping and may provide insight toward the development of novel therapeutically relevant decapping inhibitors.

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Potential Therapeutic Applications of Rna Cap Analogs

Cited by 61 publications

References 165 publications

Knockdown of eukaryotic translation initiation factor 4E suppresses cell growth and invasion, and induces apoptosis and cell cycle arrest in a human lung adenocarcinoma cell line

Knockdown of eukaryotic translation initiation factor 4E suppresses cell growth and invasion, and induces apoptosis and cell cycle arrest in a human lung adenocarcinoma cell line

Triazole-containing monophosphate mRNA cap analogs as effective translation inhibitors

Two-headed tetraphosphate cap analogs are inhibitors of the Dcp1/2 RNA decapping complex

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