RNA Targeting Therapeutics: Molecular Mechanisms of Antisense Oligonucleotides as a Therapeutic Platform

Bennett, C. Frank; Swayze, Eric E.

doi:10.1146/annurev.pharmtox.010909.105654

Cited by 1,181 publications

(1,134 citation statements)

References 217 publications

(246 reference statements)

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“…The short-lived ASO Opn effect in this study may be improved by suited oligonucleotide chemistries such as 2 0 -OH modifications, or the use of locked or peptide nucleic acids. 32,33 In variance to the ASO Opn , its complementary senseoligomer showed an unexpected antitumor effect. The anti-proliferative effect of the SO in vitro was only marginally better, but surpassed that of the ASO Opn significantly in vivo.…”

Section: Discussionmentioning

confidence: 99%

Influence of osteopontin expression on the metastatic growth of CC531 rat colorectal carcinoma cells in rat liver

et al. 2011

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Development of hepatic metastasis is responsible for most of colorectal cancer-related deaths. Osteopontin (OPN) is a small integrin-binding N-linked glycoprotein, which plays a crucial role in the formation of hepatic metastasis. This study aimed to suppress Opn expression by an antisense-oligonucleotide (ASO Opn ) to decrease liver metastasis in vivo. The effect of ASO Opn was investigated in vitro in CC531 lacZ colorectal cancer cells in comparison to sense (SO) or nonsense (NSO) oligomers, by determining mRNA and protein expression levels, as well as cell survival. For in vivo treatment, CC531 lacZ cells were intraportally inoculated into rats to compare the effects of ASO, SO and NSO oligomers, following prolonged subcutaneous administration by osmotic minipumps. The resulting CC531 lacZ tumor cell load in the liver was measured by a b-galactosidase assay. Proliferation of CC531 lacZ cells in vitro was significantly decreased after ASO Opn and SO treatment (Po0.001). Liver metastasis development was reduced as long as ASO Opn was administered, but this effect was rapidly blunted following the end of the ASO Opn administration. In contrast, administration of the SO resulted in a tumor load reduction, which surprisingly surpassed the ASO Opn effect in vivo in terms of a long-lasting metastasis suppression, which was accompanied with increased survival of the animals. Administration of the ASO Opn in rats was effective in decreasing their liver metastasis. The short-lived effect might be extended by modifications suited to increase the ASOs' half-life. In addition, there was a superior anti-metastatic effect caused by the SO, which has not been reported previously.

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Section: Discussionmentioning

confidence: 99%

Influence of osteopontin expression on the metastatic growth of CC531 rat colorectal carcinoma cells in rat liver

et al. 2011

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“…54 The ASOs used in this study were fully modified with the phosphorothioate backbone modification and had a central gap region of ten DNA nucleotides flanked on either end with three constrained ethyl (cEt) nucleotides. 32,34 The phosphorothioate modification enhances ASO metabolic stability and promotes binding to plasma and cell surface proteins, 55 which helps the ASO distribute from the site of injection to peripheral tissues.…”

Section: Discussionmentioning

confidence: 99%

Antisense Reduction of Mutant COMP Reduces Growth Plate Chondrocyte Pathology

et al. 2017

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Mutations in cartilage oligomeric matrix protein cause pseudoachondroplasia, a severe disproportionate short stature disorder. Mutant cartilage oligomeric matrix protein produces massive intracellular retention of cartilage oligomeric matrix protein, stimulating ER and oxidative stresses and inflammation, culminating in post-natal loss of growth plate chondrocytes, which compromises linear bone growth. Treatments for pseudoachondroplasia are limited because cartilage is relatively avascular and considered inaccessible. Here we report successful delivery and treatment using antisense oligonucleotide technology in our transgenic pseudoachondroplasia mouse model. We demonstrate delivery of human cartilage oligomeric matrix protein-specific antisense oligonucleotides to cartilage and reduction of cartilage oligomeric matrix protein expression, which largely alleviates pseudoachondroplasia growth plate chondrocyte pathology. One antisense oligonucleotide reduced steady-state levels of cartilage oligomeric matrix protein mRNA and dampened intracellular retention of mutant cartilage oligomeric matrix protein, leading to a reduction of inflammatory markers and cell death and partial restoration of proliferation. This novel and exciting work demonstrates that antisense-based therapy is a viable approach for treating pseudoachondroplasia and other human cartilage disorders. INTRODUCTIONTreatments of skeletal dysplasia/dwarfing conditions, including pseudoachondroplasia (PSACH), are few and fraught with problems because of the inaccessibility of chondrocytes within the cartilage matrix and the relative avascular nature of the cartilage environment. PSACH is clinically characterized by disproportionate short stature, rhizomelic shortening of the long bones, brachydactyly, and extreme joint laxity. 1,2 Joint pain in childhood and osteoarthritis (OA) in early adulthood are the most debilitating features of PSACH; only symptomatic treatments are available and have limited efficacy. 2,3 The diagnosis is made between 2-3 years of age when linear growth slows and a waddling gait develops. 1,2 Because the diagnosis, in most cases, is made post-natally, treatments aimed at resolving the pathology will have to be started immediately after diagnosis.PSACH is caused by mutations in cartilage oligomeric matrix protein (COMP), a pentameric extracellular matrix (ECM) glycoprotein abundant in musculoskeletal tissues. 4,5 Functionally, COMP facilitates type II collagen fibril assembly, enhances chondrocyte attachment in vitro, and interacts with other ECM proteins, including type II and IX collagens, matrilin 3, and SPARC. [6][7][8][9][10][11][12] In contrast, mutations in COMP cause misfolding of the protein, preventing export from the chondrocyte and resulting in massive retention within the endoplasmic reticulum (ER). 10,[13][14][15] Although this finding has long been appreciated, there was little understanding of the pathologic molecular mechanisms, which are critical to develop mechanism-driven therapeutics. To circumvent this proble...

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“…2,[4][5][6] Although most reported splicing mutations directly abolish an authentic splice site or create a novel one, an increasing number of disease-associated variations that alter splicing enhancers or silencers have been reported. 2,[7][8][9] Each nucleotide modification should be considered a potential candidate for splicing alterations, as not only intronic but also nonsense, missense and silent modifications may impact splicing. 7 Accordingly, constitutive and regulated splicing reactions are considered potential therapeutic targets and novel strategies for their correction are evolving.…”

Section: Introductionmentioning

confidence: 99%

“…Among these, antisense oligonucleotides display an exquisite specificity, being capable of distinguishing a single nucleotide-mismatch. 8 Effectively, spinal muscular atrophy is a candidate for this therapy, with promising results on patient-derived cell lines that have been already described. [9][10][11] Moreover, many antisense oligonucleotides are already in phase II/III clinical trials, namely AVI4658 (Eteplirsen), a morpholino oligonucleotide for duchenne muscular dystrophy.…”

Section: Introductionmentioning

confidence: 99%

Functional correction by antisense therapy of a splicing mutation in the GALT gene

et al. 2014

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In recent years, antisense therapy has emerged as an increasingly important therapeutic approach to tackle several genetic disorders, including inborn errors of metabolism. Intronic mutations activating cryptic splice sites are particularly amenable to antisense therapy, as the canonical splice sites remain intact, thus retaining the potential for restoring constitutive splicing. Mutational analysis of Portuguese galactosemic patients revealed the intronic variation c.820 þ 13A4G as the second most prevalent mutation, strongly suggesting its pathogenicity. The aim of this study was to functionally characterize this intronic variation, to elucidate its pathogenic molecular mechanism(s) and, ultimately, to correct it by antisense therapy. Minigene splicing assays in two distinct cell lines and patients' transcript analyses showed that the mutation activates a cryptic donor splice site, inducing an aberrant splicing of the GALT pre-mRNA, which in turn leads to a frameshift with inclusion of a premature stop codon (p.D274Gfs*17). Functional-structural studies of the recombinant wild-type and truncated GALT showed that the latter is devoid of enzymatic activity and prone to aggregation. Finally, two locked nucleic acid oligonucleotides, designed to specifically recognize the mutation, successfully restored the constitutive splicing, thus establishing a proof of concept for the application of antisense therapy as an alternative strategy for the clearly insufficient dietary treatment in classic galactosemia. European Journal of Human Genetics (2015) 23, 500-506; doi:10.1038/ejhg.2014.149; published online 23 July 2014 INTRODUCTIONOver the last years, splicing mutations emerged as an important pathogenic mechanism, underlying 10-30% of genetic diseases (HGMD Professional 2013.1). 1 Splicing accuracy depends not only on the recognition of exon-intron junctions, defined by intronic ciselements: 5 0 splice site, 3 0 splice site, branch site and poly-pyrimidine tract, 2-4 but also on more discrete elements, entitled splicing regulatory elements, which direct the splicing machinery to use the correct splice sites. Exonic and intronic splicing enhancers stimulate splicing and serve as binding sites mainly for serine/arginine-rich proteins. Exonic and intronic splicing silencers repress splicing, and often function by binding proteins from the heterogenous nuclear ribonucleoprotein family. 2,4-6 Although most reported splicing mutations directly abolish an authentic splice site or create a novel one, an increasing number of disease-associated variations that alter splicing enhancers or silencers have been reported. 2,7-9 Each nucleotide modification should be considered a potential candidate for splicing alterations, as not only intronic but also nonsense, missense and silent modifications may impact splicing. 7 Accordingly, constitutive and regulated splicing reactions are considered potential therapeutic targets and novel strategies for their correction are evolving. Among these, antisense oligonucleotides display an exquisi...

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RNA Targeting Therapeutics: Molecular Mechanisms of Antisense Oligonucleotides as a Therapeutic Platform

Cited by 1,181 publications

References 217 publications

Influence of osteopontin expression on the metastatic growth of CC531 rat colorectal carcinoma cells in rat liver

Influence of osteopontin expression on the metastatic growth of CC531 rat colorectal carcinoma cells in rat liver

Antisense Reduction of Mutant COMP Reduces Growth Plate Chondrocyte Pathology

Functional correction by antisense therapy of a splicing mutation in the GALT gene

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