Mechanisms of cellular uptake and endosomal escape of calcium-siRNA nanocomplexes

Goldshtein, Matan; Forti, Efrat; Ruvinov, Emil; Cohen, Smadar

doi:10.1016/j.ijpharm.2016.10.009

Cited by 32 publications

(46 citation statements)

References 53 publications

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“…The excess of calcium ions inside the cell could then be promptly removed by calcium pumps on the cell membrane [124]. Goldshtein et al, have also confirmed that Ca 2+ ions detach from the siRNA once in the endosomal acidic environment, escaping from the endosome and allowing protons to enter, leading to swelling and eventual rupture of the early endosome [125].…”

Section: Calcium Phosphates Nanoparticles As Non-viral Vectorsmentioning

confidence: 94%

“…Chowdhury report a ten-fold increase in transfection following the incorporation of Mg + into calcium phosphate particles when compared to calcium phosphate alone [129]. However, contradictory results were reported by Goldshtein et al, who suggested that the higher intracellular concentration of Mg 2+ would impede the cellular uptake of Mg 2+ doped calcium phosphate [125].…”

Section: Calcium Phosphates Nanoparticles As Non-viral Vectorsmentioning

confidence: 99%

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Calcium Phosphate Nanoparticles-Based Systems for RNAi Delivery: Applications in Bone Tissue Regeneration

et al. 2020

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Bone-related injury and disease constitute a significant global burden both socially and economically. Current treatments have many limitations and thus the development of new approaches for bone-related conditions is imperative. Gene therapy is an emerging approach for effective bone repair and regeneration, with notable interest in the use of RNA interference (RNAi) systems to regulate gene expression in the bone microenvironment. Calcium phosphate nanoparticles represent promising materials for use as non-viral vectors for gene therapy in bone tissue engineering applications due to their many favorable properties, including biocompatibility, osteoinductivity, osteoconductivity, and strong affinity for binding to nucleic acids. However, low transfection rates present a significant barrier to their clinical use. This article reviews the benefits of calcium phosphate nanoparticles for RNAi delivery and highlights the role of surface functionalization in increasing calcium phosphate nanoparticles stability, improving cellular uptake and increasing transfection efficiency. Currently, the underlying mechanistic principles relating to these systems and their interplay during in vivo bone formation is not wholly understood. Furthermore, the optimal microRNA targets for particular bone tissue regeneration applications are still unclear. Therefore, further research is required in order to achieve the optimal calcium phosphate nanoparticles-based systems for RNAi delivery for bone tissue regeneration.

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Section: Calcium Phosphates Nanoparticles As Non-viral Vectorsmentioning

confidence: 94%

Section: Calcium Phosphates Nanoparticles As Non-viral Vectorsmentioning

confidence: 99%

Calcium Phosphate Nanoparticles-Based Systems for RNAi Delivery: Applications in Bone Tissue Regeneration

et al. 2020

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“…One approach is to treat cells with small chemicals, such as Ca 2+ [ 13 ], chloroquine [ 14 ], and sucrose [ 15 ]. Ca 2+ may induce endosomal escape via the proton sponge effect [ 16 ], which works by causing osmotic swelling and eventual rupture of the vesicles [ 11 , 17 – 20 ]. Similarly, chloroquine can induce the vesicle rupture through protonation in acidic environment of late endosomes and lysosomes [ 11 ].…”

Section: Introductionmentioning

confidence: 99%

Distinct effects of endosomal escape and inhibition of endosomal trafficking on gene delivery via electrotransfection

et al. 2017

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A recent theory suggests that endocytosis is involved in uptake and intracellular transport of electrotransfected plasmid DNA (pDNA). The goal of the current study was to understand if approaches used previously to improve endocytosis of gene delivery vectors could be applied to enhancing electrotransfection efficiency (eTE). Results from the study showed that photochemically induced endosomal escape, which could increase poly-L-lysine (PLL)-mediated gene delivery, decreased eTE. The decrease could not be blocked by treatment of cells with endonuclease inhibitors (aurintricarboxylic acid and zinc ion) or antioxidants (L-glutamine and ascorbic acid). Chemical treatment of cells with an endosomal trafficking inhibitor that blocks endosome progression, bafilomycin A1, resulted in a significant decrease in eTE. However, treatment of cells with lysosomotropic agents (chloroquine and ammonium chloride) had little effects on eTE. These data suggested that endosomes played important roles in protecting and intracellular trafficking of electrotransfected pDNA.

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“…As efforts to develop superior targeting strategies for oligo-based therapeutics intensify, researchers are increasingly looking toward protein conjugates as delivery vehicles. Given their high affinity to cell-or tissue-specific target receptors resulting in receptor-mediated endocytosis and en-dosomal release of siRNA into the cytoplasm, mAb-siRNA conjugates present a potential delivery solution [6,[15][16][17][18][19][20]. Therapeutic development of mAb-siRNA conjugates will require sensitive and specific bioanalytical assays.…”

Section: Resultsmentioning

confidence: 99%

Quantification of siRNA-Antibody Conjugates in Biological Matrices by Triplex-Forming Oligonucleotide ELISA

Humphreys

Thayer

Campuzano

et al. 2019

Nucleic Acid Therapeutics

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The potential repertoire of short interfering RNA (siRNA) therapeutics is expanding as targeting strategies evolve. One approach to enable organ-specific delivery has been to directly conjugate siRNA to a monoclonal antibody (siRNA-mAb), analogous to antibody-drug conjugates. Detection of intact siRNA-mAb conjugates presents a bioanalytical challenge given that certain synthetic nucleotide chemical modifications and lowtemperature requirements render common oligonucleotide detection assays, such as reverse transcriptionpolymerase chain reaction, incompatible with the immunoassay component. To circumvent these issues, we developed a triplex-forming oligonucleotide ELISA using locked nucleic acid (LNA) containing oligonucleotide probes. We demonstrate that the incorporation of these LNAs allow for an enrichment and immobilization of siRNA directly conjugated to an antibody at nondenaturing temperatures. Without further requirement for extraction or amplification, we can sensitively and specifically detect intact siRNA-mAb conjugates in complex matrices such as serum and tissue homogenate.

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Mechanisms of cellular uptake and endosomal escape of calcium-siRNA nanocomplexes

Cited by 32 publications

References 53 publications

Calcium Phosphate Nanoparticles-Based Systems for RNAi Delivery: Applications in Bone Tissue Regeneration

Calcium Phosphate Nanoparticles-Based Systems for RNAi Delivery: Applications in Bone Tissue Regeneration

Distinct effects of endosomal escape and inhibition of endosomal trafficking on gene delivery via electrotransfection

Quantification of siRNA-Antibody Conjugates in Biological Matrices by Triplex-Forming Oligonucleotide ELISA

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