A novel chimeric cell-penetrating peptide with membrane-disruptive properties for efficient endosomal escape

Salomone, Fabrizio; Cardarelli, Francesco; Luca, Mariagrazia Di; Boccardi, Claudia; Nifosı̀, Riccardo; Bardi, Giuseppe; Bari, Lorenzo Di; Serresi, Michela; Beltram, Fabio

doi:10.1016/j.jconrel.2012.09.019

Cited by 120 publications

(130 citation statements)

References 46 publications

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“…S1-S7). This is consistent with reports that show that endosomal release of CPP-cargo conjugates is sometimes very inefficient, one factor that currently limits their utility for macromolecular cargo delivery (19,25,29,30). Another factor that can limit the utility of cationic CPPs is that they do not enter cells that are not capable of endocytosis.…”

Section: Discussionsupporting

confidence: 90%

“…Once endocytosed, there is the complicating question of cargo release from endosomes that has sometimes been attributed to spontaneous membrane translocation (16). As we show below, endosomal release can be very inefficient (19,25,29,30) and can effectively prevent the delivery of some endocytosed cargos to the cytosol. Given this roadblock to delivery, various approaches based on physical or osmotic lysis of endosomes have been tested to increase the efficiency of endosomal release postuptake (22,29,(31)(32)(33)(34).…”

mentioning

confidence: 82%

“…As we show below, endosomal release can be very inefficient (19,25,29,30) and can effectively prevent the delivery of some endocytosed cargos to the cytosol. Given this roadblock to delivery, various approaches based on physical or osmotic lysis of endosomes have been tested to increase the efficiency of endosomal release postuptake (22,29,(31)(32)(33)(34). Still, despite significant efforts over the last two decades, the highly cationic cell-penetrating peptides have yet to achieve their potential as delivery vehicles in the laboratory or in the clinic.…”

mentioning

confidence: 94%

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Direct Cytosolic Delivery of Polar Cargo to Cells by Spontaneous Membrane-translocating Peptides

He¹,

Kauffman²,

Fuselier³

et al. 2013

Journal of Biological Chemistry

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Background: Spontaneous membrane-translocating peptides were discovered by screening in synthetic lipid vesicles. Results: The translocating peptides carry membrane-impermeant cargos directly across cell membranes and drive systemic biodistribution in small animals. Conclusion: These peptides constitute a new class of delivery vehicle for membrane-impermeant cargos. Significance: Spontaneous membrane-translocating peptides could expand the universe of useful drugs.

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Section: Discussionsupporting

confidence: 90%

mentioning

confidence: 82%

mentioning

confidence: 94%

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Direct Cytosolic Delivery of Polar Cargo to Cells by Spontaneous Membrane-translocating Peptides

He¹,

Kauffman²,

Fuselier³

et al. 2013

Journal of Biological Chemistry

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“…Calcein, a membrane-impermeable fluorophore, was employed as a cargo tracer for cytosolic release from endosomes after endocytic uptakes by co-treated internalizing agents, because it cannot penetrate into cytosol from endosomes for itself. 35 Treatment of calcein alone with live HeLa cells did not display any significant fluorescence in the cytosol while showing very dim fluorescence in the punctate endosomal pattern (Fig. 4D).…”

Section: Intracellular Trafficking and Stability Of Internalized Cytomentioning

confidence: 90%

“…Subsequently, 50 mM of calcein was added for 2 h at 37 C. 35 Cells were then washed 3 times with PBS and fixed. Calcein fluorescence was analyzed by confocal microscopy.…”

Section: Analysis Of Calcein Release From Endosomesmentioning

confidence: 99%

A general strategy for generating intact, full-length IgG antibodies that penetrate into the cytosol of living cells

Choi¹,

Bae

Shin

et al. 2014

mAbs

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These authors contributed equally to this work.Keywords: cell-penetrating antibody, cytosolic protein targeting, cellular internalization, endosomal release, IgG antibody, intracellular trafficking, next-generation antibodyAbbreviations: IgG, immunoglobulin G; VL, light chain variable domain; VH, heavy chain variable domain; LC, light chain; HC, heavy chain; CDR, complementarity-determining region.Full-length IgG antibodies cannot cross cell membranes of living cells; this limits their use for direct targeting of cytosolic proteins. Here, we describe a general strategy for the generation of intact, full-length IgG antibodies, herein called cytotransmabs, which internalize into living cells and localize in the cytosol. We first generated a humanized light chain variable domain (VL) that could penetrate into the cytosol of living cells and was engineered for association with various subtypes of human heavy chain variable domains (VHs). When light chains with humanized VL were coexpressed with 3 heavy chains (HCs), including 2 HCs of the clinically approved adalimumab (Humira Ò ) and bevacizumab (Avastin Ò ), all 3 purified IgG antibodies were internalized into the cytoplasm of living cells. Cytotransmabs primarily internalized into living cells by the clathrin-mediated endocytic pathway through interactions with heparin sulfate proteoglycan that was expressed on the cell surface. The cytotransmabs escaped into the cytosol from early endosomes without being further transported into other cellular compartments, like the lysosomes, endoplasmic reticulum, Golgi apparatus, and nucleus. Furthermore, we generated a cytotransmab that co-localized with the targeted cytosolic protein when it was incubated with living cells, demonstrating that the cytotransmab can directly target cytosolic proteins. Internalized cytotransmabs did not show any noticeable cytotoxicity and remained in the cytosol for more than 6 h before being degraded by proteosomes. These results suggest that cytotransmabs, which efficiently enter living cells and reach the cytosolic space, will find widespread uses as research, diagnostic, and therapeutic agents.

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Interactions of Nanomaterials with Biological Systems

2018

Behaviors and Persistence of Nanomaterials in Biomedical Applications

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A novel chimeric cell-penetrating peptide with membrane-disruptive properties for efficient endosomal escape

Cited by 120 publications

References 46 publications

Direct Cytosolic Delivery of Polar Cargo to Cells by Spontaneous Membrane-translocating Peptides

Direct Cytosolic Delivery of Polar Cargo to Cells by Spontaneous Membrane-translocating Peptides

A general strategy for generating intact, full-length IgG antibodies that penetrate into the cytosol of living cells

Interactions of Nanomaterials with Biological Systems

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