Nuclease‐resistant 63‐bp trimeric si<scp>RNA</scp>s simultaneously silence three different genes in tumor cells

Gvozdeva, Olga V.; Gladkih, Daniil V.; Chernikov, Ivan V.; Meschaninova, Mariya I.; Venyaminova, Alya G.; Zenkova, Marina A.; Vlassov, Valentin V.; Chernolovskaya, E. L.

doi:10.1002/1873-3468.12927

Cited by 7 publications

(6 citation statements)

References 18 publications

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“…The observed decrease in activity can be associated with both the influence of cholesterol attachment on the thermodynamics of the duplexes and on recognition of the conjugate by proteins of the RNA-interference machinery. A more significant decrease in the activity of TsiRNA-1 may be related to the fact that this trimer, as we have demonstrated previously [8], is not processed in the cell by Dicer due to the presence of 2 OMe modifications in the vicinity of expected sites of Dicer cleavage and TsiRNA-1 acts via a Dicer-independent mechanism, which may be more sensitive to modifications of the 5 end of siRNA such as the attachment of cholesterol [9]. It was demonstrated [14] that 25-27 bp dsRNAs can be directly loaded into Ago2 and show better efficacy as compared with canonical 21-bp siRNAs; in this case, Ago2 protein may be more sensitive to the presence of chemical modifications, or it interacts with the 5 end of the duplex sense strand.…”

Section: In Vitromentioning

confidence: 58%

“…Anti-MDR1 monomeric and trimeric siRNAs and their conjugates with cholesterol connected though C6 linker (Tables 1 and 2) were synthesized as described previously [9,10] The C6 linker was selected because the monomeric siRNA conjugate with this linker showed the highest biological activity compared to the conjugates with other linkers [10]. 2'-O-Methyl modifications were introduced into nuclease-sensitive sites according to the previously developed algorithm [11] in order to prevent degradation of carrier-free siRNA in the presence of serum and in the bloodstream.…”

Section: Resultsmentioning

confidence: 99%

“…Previously, we found that 42-and 63-bp siRNAs containing 2 -OMe modifications in nuclease-sensitive sites induced more effective RNAi outcomes at lower concentrations than the classical 21-bp siRNAs without nonspecific immune effects acting in a Dicer-independent mode in cell culture after transfection [8]. Later, we designed a multimeric nuclease-resistant 63-bp trimeric small-interfering RNA (TsiRNA) comprising in one duplex the sequences of siRNAs targeting the mRNAs of the MDR1, LMP2, and LMP7 genes [9]. The results showed that such TsiRNAs are able to suppress the expression of all their target genes independently and with high efficiency, acting via a Dicer-dependent mechanism.…”

Section: Introductionmentioning

confidence: 99%

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Trimeric Small Interfering RNAs and Their Cholesterol-Containing Conjugates Exhibit Improved Accumulation in Tumors, but Dramatically Reduced Silencing Activity

et al. 2020

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Cholesterol derivatives of nuclease-resistant, anti-MDR1 small-interfering RNAs were designed to contain a 2’-OMe-modified 21-bp siRNA and a 63-bp TsiRNA in order to investigate their accumulation and silencing activity in vitro and in vivo. The results showed that increasing the length of the RNA duplex in such a conjugate increases its biological activity when delivered using a transfection agent. However, the efficiency of accumulation in human drug-resistant KB-8-5 cells during delivery in vitro in a carrier-free mode was reduced as well as efficiency of target gene silencing. TsiRNAs demonstrated a similar biodistribution in KB-8-5 xenograft tumor-bearing SCID mice with more efficient accumulation in organs and tumors than cholesterol-conjugated canonical siRNAs; however, this accumulation did not provide a silencing effect. The lack of correlation between the accumulation in the organ and the silencing activity of cholesterol conjugates of siRNAs of different lengths can be attributed to the fact that trimeric Ch-TsiRNA lags mainly in the intercellular space and does not penetrate sufficiently into the cytoplasm of the cell. Increased accumulation in the organs and in the tumor, by itself, shows that using siRNA with increased molecular weight is an effective approach to control biodistribution and delivery to the target organ.

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Section: In Vitromentioning

confidence: 58%

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Trimeric Small Interfering RNAs and Their Cholesterol-Containing Conjugates Exhibit Improved Accumulation in Tumors, but Dramatically Reduced Silencing Activity

et al. 2020

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“…The cationic liposomes 2X3-DOPE was chosen as the transfection agent for in vivo experiments, since its high transfection efficiency and low toxicity have been previously shown [ 94 , 95 ]. Time points of 4–14 days after siRNA administration and before LPS challenge were chosen based on the literature data and our previously obtained results [ 96 ]. As depicted in Figure 5 B, siTIMP1_2m decreased the level of Timp1 by 25% 8–10 days after i.n.…”

Section: Resultsmentioning

confidence: 99%

siRNA-Mediated Timp1 Silencing Inhibited the Inflammatory Phenotype during Acute Lung Injury

Chernikov

Staroseletz

Татарникова

et al. 2023

IJMS

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Acute lung injury is a complex cascade process that develops in response to various damaging factors, which can lead to acute respiratory distress syndrome. Within this study, based on bioinformatics reanalysis of available full-transcriptome data of acute lung injury induced in mice and humans by various factors, we selected a set of genes that could serve as good targets for suppressing inflammation in the lung tissue, evaluated their expression in the cells of different origins during LPS-induced inflammation, and chose the tissue inhibitor of metalloproteinase Timp1 as a promising target for suppressing inflammation. We designed an effective chemically modified anti-TIMP1 siRNA and showed that Timp1 silencing correlates with a decrease in the pro-inflammatory cytokine IL6 secretion in cultured macrophage cells and reduces the severity of LPS-induced acute lung injury in a mouse model.

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“…The extracellular barriers comprise of the degradation by nucleases, renal clearance, and uptake by the reticuloendothelial systems. Many chemical modifications have evidently enhanced the stability of siRNAs and protected against hydrolysis by the nucleases (Ueno et al, 2008(Ueno et al, , 2009Chernikov et al, 2017;Gvozdeva et al, 2018). Small size and molecular weight lower than 40 kDa render the siRNA prone to the kidney filtration process.…”

Section: General Obstacles In Sirna Deliverymentioning

confidence: 99%

Systemic Delivery of Small Interfering RNA Therapeutics: Obstacles and Advances

Chandela

Ueno

2019

RAS

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RNA interference (RNAi) has emerged as a critical tool for genetic therapeutics. Within the last two decades, there have been extensive investigations in the systemic delivery of these drugs but the obstacles in in vivo delivery have possessed a great difficulty in their administration. These developments led to the advances in systemic administration with designing and incorporation of various drug carriers, including bioconjugate systems, polymeric complexes, organic and inorganic nanoparticles. However, successful translation of these drug delivery systems for the clinical application has still been a great setback for these class of pharmaceutics. In order to improve their competency and applicability, systemic evaluation of currently available carrier systems is required. In this review, we focus on the obstacles in the systemic administration of small interfering RNAs (siRNAs) and the advances with various carrier systems to overcome the limitations of these obstacles. General obstacles in siRNA delivery have been discussed with major emphasis on the barriers at different stages of systemic administration. Then, advances in their application for targeted delivery with rationally designed carrier systems such as bioconjugates, polymeric complexes, and nanoparticles have been introduced. Lastly, we discuss the progress and state of clinical studies of siRNA therapeutics with perspectives of clinical potential.

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Nuclease‐resistant 63‐bp trimeric siRNAs simultaneously silence three different genes in tumor cells

Cited by 7 publications

References 18 publications

Trimeric Small Interfering RNAs and Their Cholesterol-Containing Conjugates Exhibit Improved Accumulation in Tumors, but Dramatically Reduced Silencing Activity

Trimeric Small Interfering RNAs and Their Cholesterol-Containing Conjugates Exhibit Improved Accumulation in Tumors, but Dramatically Reduced Silencing Activity

siRNA-Mediated Timp1 Silencing Inhibited the Inflammatory Phenotype during Acute Lung Injury

Systemic Delivery of Small Interfering RNA Therapeutics: Obstacles and Advances

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