Perfluoroaryl Bicyclic Cell‐Penetrating Peptides for Delivery of Antisense Oligonucleotides

Wolfe, Justin M.; Fadzen, Colin M.; Holden, Rebecca L.; Yao, Monica; Hanson, Gunnar J.; Pentelute, Bradley L.

doi:10.1002/ange.201801167

Cited by 22 publications

(20 citation statements)

References 23 publications

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“…However, CPP-decorated peptides are usually internalized by cells via endocytosis pathways and may suffer from entrapment within degradative or recycling vesicles during intracellular trafficking (10). These CPP-modified peptides are still susceptible to enzymatic degradation and need chemical modification such as cyclization to improve proteolytic stability and membrane permeability (11,12).…”

Section: Introductionmentioning

confidence: 99%

Fluoroalkylation promotes cytosolic peptide delivery

et al. 2020

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Cytosolic delivery of peptides remains a challenging task owing to their susceptibility to enzymatic degradation and the existence of multiple intracellular barriers. Here, we report a new strategy to address these issues by decoration of a fluorous tag on the terminal of cargo peptides. The fluorous-tagged peptides were assembled into nanostructures, efficiently internalized by cells via several endocytic pathways and released into the cytosol after endosomal escape. They were relatively stable against enzymatic degradation and showed much higher efficiency than nonfluorinated analogs and cell penetrant peptide–conjugated ones. The proposed strategy also efficiently delivered a proapoptotic peptide into specific sites in the cells and restored the function of cargo peptide after cytosolic delivery. The fluorous-tagged proapoptotic peptide efficiently inhibited tumor growth in vivo. This study provides an efficient fluorination strategy to promote the cytosolic delivery of peptides.

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Section: Introductionmentioning

confidence: 99%

Fluoroalkylation promotes cytosolic peptide delivery

et al. 2020

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“…CPPs typically range from 5 to 40 residues in length, have hydrophobic, hydrophilic, or amphipathic properties, and improve nuclear delivery of PMOs in vitro and in vivo. (17)(18)(19) Several CPPs are currently being investigated in clinical trials for delivery of therapeutic cargo, including PMO. (1,2,20,21) Cellular uptake of PMO…”

Section: Main Textmentioning

confidence: 99%

“…The library exhibited a broad activity spectrum, quantified by a high-throughput functional readout assay. (17) In this assay, HeLa cells stably transfected with an EGFP gene interrupted by a mutated intron of β-globin (IVS2-654) produce a non-fluorescent EGFP protein. Successful delivery of PMO IVS2-654 to the nucleus results in corrective splicing and EGFP synthesis.…”

Section: Assembly Of a Standardized Datasetmentioning

confidence: 99%

Interpretable Deep Learning for De Novo Design of Cell-Penetrating Abiotic Polymers

Schissel

Mohapatra

Wolfe

et al. 2020

Preprint

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There are more amino acid permutations within a 40-residue sequence than atoms on Earth. This vast chemical search space hinders the use of human learning to design functional polymers. Here we couple supervised and unsupervised deep learning with highthroughput experimentation to drive the design of high-activity, novel sequences reaching 10 kDa that deliver antisense oligonucleotides to the nucleus of cells. The models, in which natural and unnatural residues are represented as topological fingerprints, decipher and visualize sequenceactivity predictions. The new variants boost antisense activity by 50-fold, are effective in animals, are nontoxic, and can also deliver proteins into the cytosol. Machine learning can discover functional polymers that enhance cellular uptake of biotherapeutics, with significant implications toward developing therapies for currently untreatable diseases.One sentence summary: Deep learning generates de novo large functional abiotic polymers that deliver antisense oligonucleotides to the nucleus. typically occurs by energy-dependent uptake, meaning that endosomal escape presents an additional challenge. While some peptides can efficiently escape the endosome, designing a novel CPP sequence for this task is nearly impossible. In addition to the diversity of physicochemical properties of CPPs, variation in experimental design has resulted in inconsistent and sometimes contradictory datasets.(22) These inconsistencies preclude establishing sequence-activity relationships to guide the design of next-generation CPPs and can be remedied by testing PMO-CPP conjugates in a nuclear delivery-based assay that provides quantitative activity data and selects for sequences that can escape the endosome. In order to uncover CPP design principles for PMO delivery, it is necessary to have a standardized, biologically relevant dataset with which to train machine learning models.

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“…However, the application of those therapeutic antisense oligonucleotides in bacterial cells is limited by their poor transformation efficiencies without a suitable and effective carrier system. A cell-penetrating peptide is the conventional agent for transformation facilitating to antisense RNA access process 9) . Nevertheless, the inherent toxicity and poor stability of these peptides restricts the further applications 10) .…”

Section: Introductionmentioning

confidence: 99%

Nano-graphene oxide with antisense <i>vicR</i> RNA reduced exopolysaccharide synthesis and biofilm aggregation for <i>Streptococcus mutans </i>

Liu

Zhang

et al. 2020

Dent. Mater. J.

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Streptococcus mutans (S. mutans) has been proved to crucial cariogenic pathogens. Antisense vicR RNA reduced the transcription of virulence genes and lead to a reduction in biofilm formation. In the current study, a graphene-oxide plasmid transformation system was developed using interacted GO-polyethylenimine (PEI) complexes loaded with antisense vicR-expressing plasmid (GO-PEI-ASvicR). The particle size distribution and zeta potential of the GO-PEI-based ASvicR were evaluated. Quantitative real-time PCR assays were used to investigate the expression of S. mutans virulence genes. The exopolysaccharide (EPS) production in biofilm were evaluated by confocal laser scanning microscopy and anthrone method. We showed that GO-PEI could efficiently deliver the ASvicR-expressing plasmid into S. mutans cells and support excellent transcripts of ASvicR. Furthermore, GO-PEI-ASvicR significantly reduced virulent-associated gene expressions, suppressed biofilm aggregation and inhibited EPS accumulation. Our reports demonstrated that preserving nano-graphene oxide with antisense vicR RNA will be a more effective strategy for dental caries management.

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Perfluoroaryl Bicyclic Cell‐Penetrating Peptides for Delivery of Antisense Oligonucleotides

Cited by 22 publications

References 23 publications

Fluoroalkylation promotes cytosolic peptide delivery

Fluoroalkylation promotes cytosolic peptide delivery

Interpretable Deep Learning for De Novo Design of Cell-Penetrating Abiotic Polymers

Nano-graphene oxide with antisense <i>vicR</i> RNA reduced exopolysaccharide synthesis and biofilm aggregation for <i>Streptococcus mutans </i>

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