A fusogenic peptide from a sea urchin fertilization protein promotes intracellular delivery of biomacromolecules by facilitating endosomal escape

Niikura, Keisuke; Horisawa, Kenichi; Doi, Nobuhide

doi:10.1016/j.jconrel.2015.06.020

Cited by 22 publications

(16 citation statements)

References 36 publications

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“…The advantage of FP-fused antibodies over FP-fused eGFP is that allows target cell-specific intracellular delivery of various biomacromolecules. However, their activities of these fusion antibodies were somewhat low compared with that of FP-fused eGFP (21). One reason for this disadvantage may be that most of antibodies remain bound to antigens after internalization by endocytosis, likely reducing the endosomal escape efficiency.…”

Section: Discussionmentioning

confidence: 99%

“…1A). The B18 and B55 peptides are FPs that facilitated the endosomal escape of cargo proteins in HeLa cells as shown in our previous study (21). The TAT, a CPP derived from the HIV-1 virus, were also used to compare the functions of the FPs with a CPP.…”

Section: Preparation Of Fp-fused Scfv Proteins and Confirmation Of Thmentioning

confidence: 93%

“…The first is a cis method that involves 'direct (covalent) conjugation' of FPs to cargoes as described above, and the second is a trans method that involves 'simple co-incubation' of FPs with cargoes or other membrane-disruptive reagents (32). We recently reported that an eGFP-fused B55 peptide facilitated the endosomal escape not only of the eGFP-fused protein itself (via the cis method) but also of various biomacromolecules co-administered with the eGFP-fused B55 (via the trans method) (21). Thus, we reasoned that aEGFR[scFv]-B55 might also facilitated the endosomal escape of co-administered molecules in trans.…”

Section: Estimation Of the Endosomal Escape Efficiency Of Fpfused Scfmentioning

confidence: 99%

“…Because FPs exert membrane-disrupting activities through pH-dependent conformational changes (19,20), they are expected to facilitate endosomal escape at acidic pH. We found recently that the FPs, B18 and B55, which are derived from bindin, a sea urchin gamete recognition protein, facilitated the endosomal escape of FP-fused enhanced green fluorescent protein (eGFP) and of co-administered cargos such as dextrans (21). In this study, we produced B18-and B55-fused aEGFR (epidermal growth factor receptor) single-chain Fvs (aEGFR[scFv]-B18 and -B55).…”

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confidence: 99%

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Endosomal escape efficiency of fusogenic B18 and B55 peptides fused with anti-EGFR single chain Fv as estimated by nuclear translocation

2015

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Release 2015;212:85-93]. In this study, we constructed FP-fused anti-epidermal growth factor receptor (EGFR) single-chain Fv (αEGFR[scFv]) proteins and evaluated their endosomal escape efficiency by utilizing a nuclear localization signal). When the FP-fused αEGFR[scFv] proteins were incubated with A431 cells, the estimated endosomal escape efficiency of αEGFR[scFv]-B18 was significantly higher than that of αEGFR[scFv] alone, suggesting that the B18 peptide facilitates endosomal escape of the conjugated scFv in cis. Moreover, αEGFR[scFv]-B55 promoted the intracellular uptake of co-administered eGFP and dextrans in trans. These results imply that B18- and B55-fused antibodies may be useful for the cell-specific intracellular delivery of biomacromolecules.

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Section: Discussionmentioning

confidence: 99%

Section: Preparation Of Fp-fused Scfv Proteins and Confirmation Of Thmentioning

confidence: 93%

Section: Estimation Of the Endosomal Escape Efficiency Of Fpfused Scfmentioning

confidence: 99%

mentioning

confidence: 99%

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Endosomal escape efficiency of fusogenic B18 and B55 peptides fused with anti-EGFR single chain Fv as estimated by nuclear translocation

2015

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“…Endosome is the main bottleneck in intracellular gene delivery path. During the last few decades, several de novo designed or biologically derived endosomal release peptides have been reported 30 37 53 54 . However, the complicated mechanisms underlying endosome release and the inability of these peptides to show their actual activities, makes it more difficult to have a preference for selecting the most efficient and the most effective ones.…”

Section: Discussionmentioning

confidence: 99%

Nano-biomimetic carriers are implicated in mechanistic evaluation of intracellular gene delivery

Alipour

Hosseinkhani

Sheikhnejad³

et al. 2017

Sci Rep

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Several tissue specific non-viral carriers have been developed for gene delivery purposes. However, the inability to escape endosomes, undermines the efficacy of these carriers. Researchers inspired by HIV and influenza virus, have randomly used Gp41 and H5WYG fusogenic peptides in several gene delivery systems without any rational preference. Here for the first time, we have genetically engineered two Nano-biomimetic carriers composed of either HWYG (HNH) or Gp41 (GNH) that precisely provide identical conditions for the study and evaluation of these fusogenic peptides. The luciferase assay demonstrated a two-fold higher transfection efficiency of HNH compared to GNH. These nanocarriers also displayed equivalent properties in terms of DNA binding ability and DNA protection against serum nucleases and formed similar nanoparticles in terms of surface charge and size. Interestingly, hemolysis and cellular analysis demonstrated both of nanoparticles internalized into cells in similar rate and escaped from endosome with different efficiency. Furthermore, the structural analysis revealed the mechanisms responsible for the superior endosomal escaping capability of H5WYG. In conclusion, this study describes the rationale for using H5WYG peptide to deliver nucleic acids and suggests that using nano-biomimetic carriers to screen different endosomal release peptides, improves gene delivery significantly.

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New generation of cell‐penetrating peptides: Functionality and potential clinical application

Reißmann

Filatova

2021

Journal of Peptide Science

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Cell‐penetrating peptides (CPPs) can transport various cargoes through membranes of live cells. Since the first generations of CPPs suffered from insufficient cell and tissue selectivity, stability against proteases, and escape from endosomes, a new generation of peptides, with optimized properties, was developed. These are either derived from natural sources or created through the combination of multivalent structures. The second method allows achieving high internalization efficiency, high cell and tissue selectivity, and release from endosomes via hybrid structures, combining sequences for endosomal release, homing sequences, and sequences for activation at the target tissue and for local delivery of cargoes. CPPs with innate tumor selectivity include azurin, crotamine, maurocalcine, lycosin‐I, buffalo cathelicidin, and peptide CB5005. Some of them can penetrate the membranes of live cells and influence intracellular signaling pathways, thereby exerting cytotoxic effects against tumor cells. To obtain multilayer penetration and stabilization against proteolytic degradation, as well as for better handling, CPPs are often conjugated to nanoparticles. A special problem for tumor treatment is the efficiency of drug transport through three‐dimensional cell cultures. Therefore, the capability of CPPs to deliver the drug even to the innermost tissues is of crucial importance. Notably, the ability of certain CPPs to penetrate barriers such as skin, the blood‐brain barrier (BBB), and cornea or conjunctiva of eyes enabled the replacement of dangerous and painful injections with soothing sprays, creams, and drops. However, it is difficult to rank the efficacy of CPPs because transport efficiency and tissue selectivity depend not only on the CPP itself but also on the target tissue or organ, as well as on the cargo and method of CPP‐cargo coupling. Therefore, the present review describes some examples of new‐generation CPPs and aims to provide advice on how to find or create the right CPP for a given task.

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A fusogenic peptide from a sea urchin fertilization protein promotes intracellular delivery of biomacromolecules by facilitating endosomal escape

Cited by 22 publications

References 36 publications

Endosomal escape efficiency of fusogenic B18 and B55 peptides fused with anti-EGFR single chain Fv as estimated by nuclear translocation

Endosomal escape efficiency of fusogenic B18 and B55 peptides fused with anti-EGFR single chain Fv as estimated by nuclear translocation

Nano-biomimetic carriers are implicated in mechanistic evaluation of intracellular gene delivery

New generation of cell‐penetrating peptides: Functionality and potential clinical application

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