Structural Modifications of siRNA Improve Its Performance In Vivo

Chernikov, Ivan V.; Ponomareva, Ulyana A.; Chernolovskaya, E. L.

doi:10.3390/ijms24020956

Cited by 17 publications

(5 citation statements)

References 91 publications

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“…Still, in complex neurodegenerative diseases, single-gene targeting may be insufficient. Further, multi-targeting has been studied extensively systemically (37) but has not yet been successful in the CNS. To address the need for a multi-targeting scaffold in the CNS, we used a linear synthesis scheme for di-valent siRNA.…”

Section: Discussionmentioning

confidence: 99%

A programmable dual-targeting di-valent siRNA scaffold supports potent multi-gene modulation in the central nervous system

Belgrad,

Tang,

Hildebrand

et al. 2023

Preprint

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Di-valent short interfering RNA (siRNA) is a promising therapeutic modality that enables sequence-specific modulation of a single target gene in the central nervous system (CNS). To treat complex neurodegenerative disorders, where pathogenesis is driven by multiple genes or pathways, di-valent siRNA must be able to silence multiple target genes simultaneously. Here we present a framework for designing unimolecular “dual-targeting” di-valent siRNAs capable of co-silencing two genes in the CNS. We reconfigured di-valent siRNA – in which two identical, linked siRNAs are made concurrently – to create linear di-valent siRNA – where two siRNAs are made sequentially attached by a covalent linker. This linear configuration, synthesized using commercially available reagents, enables incorporation of two different siRNAs to silence two different targets. We demonstrate that this dual-targeting di-valent siRNA is fully functional in the CNS of mice, supporting at least two months of maximal target silencing. Dual-targeting di-valent siRNA is highly programmable, enabling simultaneous modulation of two different disease-relevant gene pairs (e.g., Huntington’s disease:MSH3andHTT; Alzheimer’s disease:APOEandJAK1) with similar potency to a mixture of single-targeting di-valent siRNAs against each gene. This work potentiates CNS modulation of virtually any pair of disease-related targets using a simple unimolecular siRNA.

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Section: Discussionmentioning

confidence: 99%

A programmable dual-targeting di-valent siRNA scaffold supports potent multi-gene modulation in the central nervous system

Belgrad,

Tang,

Hildebrand

et al. 2023

Preprint

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show abstract

“…2'OMe modification of the ribose provides a bulkier option than 2'F modification and provides better resistance to degradation by nucleases [21]. Too many of the natural base 2'OMe modifications however can diminish siRNA activity against its target, but the modifications can also suppress siRNA-driven innate immune activation, enhance specificity for the target (increasing activity), and reduce toxicity due to off-target mediated effects [22].…”

Section: Rationale For Chemically Modified Bases In Sirnamentioning

confidence: 99%

“…We didn't perform a thorough evaluation of base changes but made rational changes to the siRNA to incorporate these modifications. For example, we selected bases 9-11 on the sense strand to be modified with 2'F (since this corresponds to the cleavage location of the cognate mRNA [21,23]). We also made the 14th base in the antisense strand a 2'F since this position does not tolerate 2'OMe changes [24].…”

Section: Rationale For Chemically Modified Bases In Sirnamentioning

confidence: 99%

CHK1 siRNA Containing Gemcitabine and Chemically Modified Nucleotides Shows Antitumoral Activity in Vitro and in Vivo

2024

J Oncol Res Ther

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Gemcitabine was shown to augment anti-cancer activity when combined with small molecule CHK1 inhibitors. We had previously demonstrated that we could incorporate gemcitabine molecules into the backbone of an siRNA targeting CHK1 and studied this drugs effects in pancreatic cancer cells [1]. We had optimized the number of gemcitabines to improve efficacy in the combination where the CHK1 siRNA was unmodified [1]. In this manuscript, we extend these studies by using chemically modified siRNAs using the same siRNA sequence targeting CHK1. We varied the 2'-Fluoro and 2'-O-Methyl modification patterns within the sequence, with gemcitabines in the same locations within the same siRNA sequence. We show that adding gemcitabines into the siRNA can demonstrate improved potency, not only against pancreatic cancer cell models but also against lung cancer, ovarian cancer, triple negative breast cancer and colon cancer.We formulated the different versions of CHK1 siRNA + gemcitabine into polypeptide nanoparticles with good control over size (~82-88nm) and zeta potential (34-36mV). When delivered intravenously to mice bearing a pancreatic xenograft tumor, the cocktail showed a reduction in tumor burden. The product demonstrated inhibitory effects in vivo, with no effect on the body weights of the animals.We further examined the ability to improve on the potency of this reagent by inclusion of a second siRNA (targeting WEE1) and demonstrate that, in vitro, the pancreatic cancer cell line, MiaPaca2, is particularly sensitive to a combo of CHK1 siRNA plus gemcitabine together with a second siRNA against WEE1.

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“…The in vivo delivery of siRNA requires overcoming blood, tissue, cellular, and intracellular barriers in order to reach the target site and exert gene silencing. Blood stability, targeting, tumor permeability, and endosome escape ability are key properties of siRNA carriers [ 12 ]. Many nanocarrier systems have been developed for siRNA delivery to tumor tissues, and lipid nanoparticles (LNPs) remain one of the most attractive types of siRNA carriers [ 13 ].…”

Section: Introductionmentioning

confidence: 99%

The Efficacy and Safety of Polyethylene Glycol Cholesterol- and Tocopherol Polyethylene Glycol 1000 Succinate-Modified Transforming Growth Factor β1 Small Interfering RNA Lipid Nanoparticles in the Treatment of Paclitaxel-Resistant Non-Small-Cell Lung Cancer

Zeng,

et al. 2024

Pharmaceutics

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The aim of this study was to explore the efficacy and safety of TGFβ1 siRNA lipid nanoparticles (LNPs) modified with different PEG derivatives (PEG5000 cholesterol, abbreviated as CE; tocopherol polyethylene glycol 1000 succinate, abbreviated as TPGS) in the treatment of paclitaxel-resistant non-small-cell lung cancer. Three kinds of TGFβ1 siRNA LNPs were prepared via microfluidics technology, using different PEG derivatives and dosages (CE1.5, CE2.5, TPGS2.5) as variables. Their particle size, zeta potential, contents, and encapsulation efficiencies were determined. The inhibition of TGFβ1 mRNA and protein expression and the effects of the three kinds of LNPs on the proliferation of paclitaxel-resistant non-small-cell lung cancer cells (A549/T cell) were characterized. The distributions of the three siRNA LNPs in nude mice bearing A549/T tumors, especially at the tumor site, were observed using in vivo mouse imaging technology, and their corresponding efficacies were evaluated. The average particle size of the three kinds of TGFβ1 siRNA LNPs was about 70–80 nm, and they were capable of charge flipping. All three siRNA LNPs could effectively inhibit the expression of TGFβ1 mRNA and protein in A549/T cells and inhibit the proliferation of A549/T cells in vitro. The results of in vivo mice imaging showed that the three kinds of siRNA LNPs, when labeled with cypate, retain strong fluorescence in the tumor at 24 h. The pharmacodynamic results, such as for relative tumor volumes and tumor inhibition rates, reveal that TGFβ1 siRNA LNPs modified with CE1.5, CE2.5, or TPGS2.5 can be used to effectively treat paclitaxel-resistant lung adenocarcinoma. The histopathological results showed that the three kinds of LNPs have a certain toxicity but are relatively safe compared to common forms of chemotherapy such as cabazitaxel. TGFβ1 siRNA LNPs modified with CE1.5, CE2.5, and TPGS2.5 can inhibit TGFβ1 mRNA and protein expression in A549/T cells in vitro and can accumulate and play a role in the tumor tissue of nude mice, features that can be exploited for treating paclitaxel-resistant lung adenocarcinoma.

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Structural Modifications of siRNA Improve Its Performance In Vivo

Cited by 17 publications

References 91 publications

A programmable dual-targeting di-valent siRNA scaffold supports potent multi-gene modulation in the central nervous system

A programmable dual-targeting di-valent siRNA scaffold supports potent multi-gene modulation in the central nervous system

CHK1 siRNA Containing Gemcitabine and Chemically Modified Nucleotides Shows Antitumoral Activity in Vitro and in Vivo

The Efficacy and Safety of Polyethylene Glycol Cholesterol- and Tocopherol Polyethylene Glycol 1000 Succinate-Modified Transforming Growth Factor β1 Small Interfering RNA Lipid Nanoparticles in the Treatment of Paclitaxel-Resistant Non-Small-Cell Lung Cancer

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