Systematic chemical modifications of single stranded siRNAs significantly improved CTNNB1 mRNA silencing

Chang, Wonsuk; Pei, Yi; Guidry, Erin N.; Zewge, Daniel; Parish, Craig A.; Sherer, Edward C.; DiMuzio, Jillian; Zhang, Hangchun; South, Victoria J.; Strapps, Walter; Sepp‐Lorenzino, Laura; Colletti, Steven L.; Stanton, Matthew

doi:10.1016/j.bmcl.2016.07.064

Cited by 7 publications

(8 citation statements)

References 23 publications

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“…Stapled peptides for β-catenin have been reported [ 34 ], but their activity in relevant tumor models in vivo has not been demonstrated. Therapeutic siRNA represents an appealing approach to directly inhibit β-catenin [ 35 , 36 ]; however, convincing evidence of RNAi delivery to human tumors in a clinical setting is still lacking. Given the unequivocally important contribution of β-catenin to pathogenesis of HCC and other human malignancies, novel strategies for direct targeting of this oncogene need to be developed.…”

Section: Discussionmentioning

confidence: 99%

Oncogenic dependency on β-catenin in liver cancer cell lines correlates with pathway activation

et al. 2017

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Hepatocellular carcinoma (HCC) represents a serious public health challenge with few therapeutic options available to cancer patients.Wnt/β-catenin pathway is thought to play a significant role in HCC pathogenesis. In this study, we confirmed high frequency of CTNNB1 (β-catenin) mutations in two independent cohorts of HCC patients and demonstrated significant upregulation of β-catenin protein in the overwhelming majority of HCC patient samples, patient-derived xenografts (PDX) and established cell lines. Using genetic tools validated for target specificity through phenotypic rescue experiments, we went on to investigate oncogenic dependency on β-catenin in an extensive collection of human HCC cells lines. Our results demonstrate that dependency on β-catenin generally tracks with its activation status. HCC cell lines that harbored activating mutations in CTNNB1 or displayed elevated levels of non-phosphorylated (active) β-catenin were significantly more sensitive to β-catenin siRNA treatment than cell lines with wild-type CTNNB1 and lower active β-catenin. Finally, significant therapeutic benefit of β-catenin knock-down was demonstrated in established HCC tumor xenografts using doxycycline-inducible shRNA system. β-catenin downregulation and tumor growth inhibition was associated with reduction in AXIN2, direct transcriptional target of β-catenin, and decreased cancer cell proliferation as measured by Ki67 staining. Taken together, our data highlight fundamental importance of aberrant β-catenin signaling in the maintenance of oncogenic phenotype in HCC.

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Section: Discussionmentioning

confidence: 99%

Oncogenic dependency on β-catenin in liver cancer cell lines correlates with pathway activation

et al. 2017

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“…In spite of this, some other strategies also achieve promising activity. ( Haringsma et al, 2012 ; Chang et al, 2016 ). LNP is a mixed nanocarrier that contains ionizable cationic lipid, DSPE-PEG, and cholesterol.…”

Section: Introductionmentioning

confidence: 99%

Optimization in Chemical Modification of Single-Stranded siRNA Encapsulated by Neutral Cytidinyl/Cationic Lipids

Wang

Zhou

et al. 2022

Front. Chem.

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Single-stranded siRNA (ss-siRNA) refers to the antisense strand of siRNA, which plays the role of gene silencing. Since single-stranded RNA is unstable, the present study employed a homemade neutral cytidinyl/cationic lipid delivery system and chemical modifications to improve its stability. The results showed that with the aid of mixed lipids, ss-siRNA could knock down 40% of target mRNA at 25 nM. With 2ʹ-F/2ʹ-OMe, phosphorothioate and 5ʹ-terminal phosphorylation, the optimized ss-siRNA could knock down 80% of target mRNA at 25 nM. Further knocking down AGO2, the ss-siRNAs could not effectively silence target mRNAs. Analysis of the biodistribution in vivo showed that ss-siRNA was less durable than siRNA, but spread more quickly to tissues. This study provides a safe and efficient ss-siRNA delivery and modification strategy, which lays the foundation for future works.

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“…One option for increasing the value of the RNAi mechanism for therapy is to combine the strengths of the duplex RNA and ASO approaches. This goal can be achieved using single-stranded silencing RNAs (ss-siRNAs) [5][6][7][8][9][10][11][12][13][14][15]. ss-siRNAs are chemically modified single-stranded oligonucleotides that silence gene expression through RNAi in cell culture and animals.…”

Section: Introductionmentioning

confidence: 99%

“…By contrast, much more fundamental research is needed to understand the potential of ss-siRNAs as an approach for drug discovery. This research will involve medicinal chemistry aimed at improving the characteristics of ss-siRNAs [13,14] and studies that test the boundaries of ss-siRNA function on varied gene targets.…”

Section: Introductionmentioning

confidence: 99%

“…We had previously demonstrated that duplex RNAs complementary to the hexanucleotide repeat could block both the sense and antisense transcripts, causing a substantial reduction in nuclear RNA foci [19]. We had also previously observed that ss-siRNAs could inhibit expression of genes (Huntington, atrophin-1, and ataxin-3) containing expanded CAG trinucleotide repeats [7,8,14,15]. Our goal here is to test the hypothesis that ss-siRNAs can also block the sense and antisense repeats within the c9orf72 gene.…”

Section: Introductionmentioning

confidence: 99%

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Recognition of c9orf72 Mutant RNA by Single-Stranded Silencing RNAs

Rigo

Prakash

et al. 2017

Nucleic Acid Therapeutics

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Mutations within the chromosome 9 open reading frame 72 (c9orf72) gene are associated with both familial amyotrophic lateral sclerosis and frontotemporal dementia. The mutation leads to an expanded GGGGCC hexanucleotide repeat within the first intron of c9orf72 and an expanded CCCCGG repeat within a corresponding antisense transcript. Both the mutant intronic and antisense RNAs have been implicated in disease. We have previously reported that duplex RNAs complementary to the repeats can recognize disease-causing RNA and block detection of nuclear foci formed by the mutant transcripts. Here, we test the hypothesis that inhibition can also be achieved by single-stranded silencing RNAs (ss-siRNAs). ss-siRNAs are single-stranded antisense oligonucleotides (ASOs) that function through RNAi interference (RNAi) to silence gene expression. ss-siRNAs can block the expanded repeats within both intronic RNA and the antisense transcripts. Inhibition is more potent than by analogous duplex RNAs. Our data suggest that the potent effects on foci are caused by a combination of mechanisms including RNAi and direct binding of the ss-siRNA to the target transcripts. These findings reinforce the suggestion that ss-siRNAs combine the favorable properties of duplex RNA and single-stranded ASOs.

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Systematic chemical modifications of single stranded siRNAs significantly improved CTNNB1 mRNA silencing

Cited by 7 publications

References 23 publications

Oncogenic dependency on β-catenin in liver cancer cell lines correlates with pathway activation

Oncogenic dependency on β-catenin in liver cancer cell lines correlates with pathway activation

Optimization in Chemical Modification of Single-Stranded siRNA Encapsulated by Neutral Cytidinyl/Cationic Lipids

Recognition of c9orf72 Mutant RNA by Single-Stranded Silencing RNAs

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