A Cancer Specific Cell-Penetrating Peptide, BR2, for the Efficient Delivery of an scFv into Cancer Cells

Lim, Kyung Taek; Sung, Bong Hyun; Shin, Jae Yong; Lee, Young Woong; Kim, Da Jung; Yang, Kyung Seok; Kim, Sun Chang

doi:10.1371/journal.pone.0066084

Cited by 103 publications

(79 citation statements)

References 43 publications

(38 reference statements)

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“…E.g. the efficiency of cargo introduction by TAT into HT29 [34] and BR2 into HCT116 cells was reported [35]. Our study is the first focused on the effects of TP, TP10 and their biotinylated derivates on cell viability and large protein cargo delivery into early-staged HT29 and metastatic colorectal cancer HCT116 cells.…”

Section: Discussionmentioning

confidence: 79%

Protein and siRNA delivery by transportan and transportan 10 into colorectal cancer cell lines

Wierzbicki

Kogut-Wierzbicka

Ruczyński

et al. 2015

Folia Histochem Cytobiol.

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Introduction. Cell penetrating peptides (CPPs) have the ability to translocate through cell membranes with high efficiency and therefore can introduce biological agents with pharmaceutical properties into the cell. Transportan (TP) and its shorter analog transportan 10 (TP10) are among the best studied CPPs, however, their effects on viability of and cargo introduction into colorectal cancer (CRC) cells have yet not been investigated. The aim of our study was to evaluate the cytotoxic effects of TP and TP10 on representative CRC lines and the efficiency of protein (streptavidin) and siRNA cargo delivery by TP-biotinylated derivatives (TP-biot). Material and methods. HT29 (early stage CRC model) and HCT116 (metastatic CRC model) cell lines were incubated with TP, TP10, TP-biot1, TP-biot13 and TP10-biot1. The effects of studied CPPs on cell viability and cell cycle were assessed by MTT and annexin V assays. The uptake of streptavidin-FITC complex into cells was determined by flow cytometry and fluorescence microscopy, with the inhibition of cellular vesicle trafficking by brefeldin A. The efficiency of siRNA for SASH1 gene delivery was measured by quantitative PCR (qPCR). Results. Since up to 10 µM concentrations of each CPP showed no significant cytotoxic effect, the concentrations of 0.5-5 µM were used for further analyses. Within this concentration range none of the studied CPPs affected cell viability and cell cycle. The efficient and endocytosis-independent introduction of streptavidin-FITC complex into cells was observed for TP10-biot1 and TP-biot1 with the cytoplasmic location of the fluorescent cargo; decreased SASH1 mRNA level was noticed with the use of siRNA and analyzed CPPs. Conclusions. We conclude that TP, TP10 and their biotinylated derivatives can be used as efficient delivery vehicles of small and large cargoes into CRC cells.

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Section: Discussionmentioning

confidence: 79%

Protein and siRNA delivery by transportan and transportan 10 into colorectal cancer cell lines

Wierzbicki

Kogut-Wierzbicka

Ruczyński

et al. 2015

Folia Histochem Cytobiol.

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“…For example, penetratin peptides infused into mice brain have been shown to preferentially bind to a-2,8-polysialic acid on the neuronal cell surface, with selective uptake in neurons [29]. In another study, Lim et al [30] showed a specific binding of CPPs to cancer cells. These researchers showed that the cell-penetrating motif of the anticancer peptide bufforin IIb targets cancer cells by specifically interacting with gangliosides, with subsequent internalization by lipid-mediated pinocytosis.…”

Section: The Inverted Micelle Modelmentioning

confidence: 99%

Cell-penetrating peptides: strategies for anticancer treatment

Raucher

Ryu

2015

Trends in Molecular Medicine

196

159

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“…From this perspective and despite the difficulties in targeting and delivering these molecules into specific cell cytosols, we must at least consider the therapeutic potential of antibodies targeting cytosolic domains. Although delivery of these molecules is a major obstacle at this time for their use as therapeutic agents, development of technologies for the cytosolic delivery of large protein molecules is an important focus of research (35)(36)(37)(38), and it is reasonable to expect that, in the future, solutions to this problem will be found.…”

Section: Discussionmentioning

confidence: 99%

Enhancement of hERG channel activity by scFv antibody fragments targeted to the PAS domain

Harley

Starek

Jones

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

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The human human ether-à-go-go-related gene (hERG) potassium channel plays a critical role in the repolarization of the cardiac action potential. Changes in hERG channel function underlie long QT syndrome (LQTS) and are associated with cardiac arrhythmias and sudden death. A striking feature of this channel and KCNH channels in general is the presence of an N-terminal Per-Arnt-Sim (PAS) domain. In other proteins, PAS domains bind ligands and modulate effector domains. However, the PAS domains of KCNH channels are orphan receptors. We have uncovered a family of positive modulators of hERG that specifically bind to the PAS domain. We generated two single-chain variable fragments (scFvs) that recognize different epitopes on the PAS domain. Both antibodies increase the rate of deactivation but have different effects on channel activation and inactivation. Importantly, we show that both antibodies, on binding to the PAS domain, increase the total amount of current that permeates the channel during a ventricular action potential and significantly reduce the action potential duration recorded in human cardiomyocytes. Overall, these molecules constitute a previously unidentified class of positive modulators and establish that allosteric modulation of hERG channel function through ligand binding to the PAS domain can be attained.he human ether-à-go-go-related gene (hERG; or KCNH2) encodes the voltage-gated potassium channel that conducts I Kr (delayed rectifier potassium current), a critical cardiac repolarizing current (1, 2). Mutations in hERG (3) or channel block can cause long QT syndrome (LQTS) and catastrophic ventricular arrhythmias (reviewed in ref. 4). Because most drugs causing acquired LQTS block I Kr channels (5), lead compounds in development are counterscreened using a hERG cell-based safety test (6) to reduce the incidence of sudden cardiac death caused by off-target drug effects (7).A defining feature of hERG and its relatives in the potassium channel family, KCNH (8), is a large cytoplasmic region with an N-terminal Per-Arnt-Sim (PAS) domain (9). Within the hERG PAS domain, there are two functionally and structurally distinct regions: the PAS-Cap (residues 1-25) and the globular region (residues 26-135). NMR and crystallographic structures of the isolated PAS domain show that the PAS-Cap region is partially unstructured (10-12). The globular region interacts with a C-terminal domain, which, because of sequence homology with cyclic nucleotide binding domains, is termed the cyclic nucleotide binding homology (CNBh) domain (13,14). Truncations and mutations of the PAS-Cap and PAS clearly affect the gating mechanisms of the channel, which is particularly apparent in the deactivation kinetics (9,(15)(16)(17)(18). PAS domains within the tetrameric hERG channels exert a suppressive effect by slowing channel activation and recovery from inactivation, the two parameters that predominantly determine current amplitude during an action potential (AP) (19). These properties are genetically tuned in native I Kr produce...

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A Cancer Specific Cell-Penetrating Peptide, BR2, for the Efficient Delivery of an scFv into Cancer Cells

Cited by 103 publications

References 43 publications

Protein and siRNA delivery by transportan and transportan 10 into colorectal cancer cell lines

Protein and siRNA delivery by transportan and transportan 10 into colorectal cancer cell lines

Cell-penetrating peptides: strategies for anticancer treatment

Enhancement of hERG channel activity by scFv antibody fragments targeted to the PAS domain

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