Highly Efficient Delivery of Functional Cargoes by a Novel Cell-Penetrating Peptide Derived from SP140-Like Protein

Hu, Wang; Ma, Jie-Lan; Yang, Ying; Zeng, Fanhui; Liu, Changbai

doi:10.1021/acs.bioconjchem.6b00161

Cited by 28 publications

(27 citation statements)

References 37 publications

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“…To further investigate the penetration of Dot1l peptide, the interaction between Dot1l peptide and membrane was predicted using PPM web server and MCPep server. As our previous paper has shown, well-known CPP-TAT [9], hPP3 [19], and hPP10 [20,26] can partially be inserted into the membrane predicted by the PPM web server [27], similar to Dot1l peptide's weak property of membrane insertion ( Figure 4A). The MCPep server, a computational tool for the prediction of peptide (secondary structure) occurrence in lipid bilayers and aqueous environments, was used for further prediction of interactions between Dot1l peptide and membrane.…”

Section: Dot1l Peptide-membrane Interaction Predictionsupporting

confidence: 53%

“…To improve CPP-based DNA delivery ability, various approaches have been developed, such as optimization of delivery conditions and peptide modification [14,15]. While poor efficiency is still the major disadvantage of their application, some penetration enhancers have been identified [9,16,17], creating interest in identifying CPPs and attracting the attention of researchers in the field of drug delivery [6,[18][19][20].…”

Section: Introductionmentioning

confidence: 99%

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Intracellular Delivery of DNA and Protein by a Novel Cell-Permeable Peptide Derived from DOT1L

et al. 2020

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Cellular uptake and intracellular release efficiency of biomacromolecules is low because of hurdles in the cell membrane that result in limited access to intra-cellular targets with few functional effects. Cell-penetrating peptides (CPPs) act as cargo delivery vehicles to promote therapeutic molecule translocation. Here, we describe the novel CPP-Dot1l that not only penetrates by itself, but also mediates cargo translocation in cultured cells, as confirmed by fluorescence microscopy and fluorescence spectrophotometry. We conducted cytotoxicity assays and safety evaluations, and determined peptide-membrane interactions to understand the possible pathway for cargo translocation. Additional nucleic acid and covalently conjugated green fluorescence protein (GFP) studies mediated by CPP-Dot1l were conducted to show functional delivery potential. Results indicate that CPP-Dot1l is a novel and effective CPP due to its good penetrating properties in different cell lines and its ability to enter cells in a concentration-dependent manner. Its penetration efficiency can be prompted by DMSO pretreatment. In addition, not only can it mediate plasmid delivery, but CPP-Dot1l can also deliver GFP protein into cytosol. In conclusion, the findings of this study showed CPP-Dot1l is an attractive pharmaceutical and biochemical tool for future drug, regenerative medicine, cell therapy, gene therapy, and gene editing-based therapy development.

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Section: Dot1l Peptide-membrane Interaction Predictionsupporting

confidence: 53%

Section: Introductionmentioning

confidence: 99%

Intracellular Delivery of DNA and Protein by a Novel Cell-Permeable Peptide Derived from DOT1L

et al. 2020

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“…Earlier studies have reported the designing of CPP from proteins such as heparin-binding motif of human eosinophil cationic protein, human nuclear body protein, SP140-like protein, and 12 isoforms of annexin, a family of membraneinteracting human proteins. [22][23][24] However, there is no report of using the total proteome to design a CPP which would be tissue specific. In this context, we have attempted to design a corneal-specific CPP using subtractive proteomic approach.…”

Section: Discussionmentioning

confidence: 99%

Designing and enhancing the antifungal activity of corneal specific cell penetrating peptide using gelatin hydrogel delivery system

Amit¹,

Muralikumar²,

Janaki³

et al. 2019

IJN

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BackgroundFungal keratitis is a major cause of corneal blindness accounting for more than one-third of microbiologically proven cases. The management of fungal keratitis is through topical or systemic antifungal medications alone or in combination with surgical treatment. Topical medications such as natamycin and voriconazole pose major challenges due to poor penetration across the corneal epithelium. To address the issue various carrier molecules like nanoparticles, lipid vesicles, and cell penetrating peptides were explored. But the major drawback such as non-specificity and lack of bioavailability remains.PurposeIn this study, we have attempted to design corneal specific cell penetrating peptide using subtractive proteomic approach from the published literature and tried to improve its bioavailability through gelatin hydrogel delivery system.Material and MethodsUsing subtractive proteomic approach two peptides VRF005 and VRF007 were identified on the basis of solubility, cell permeability and amphipathicity. The peptides were modeled for three-dimensional structure and simulated for membrane penetration. The peptides were characterized using circular dichroism spectroscopy, dynamic light scattering and native polyacrylamide gel electrophoresis. Further uptake studies were performed on primary corneal epithelial cells and the stability was analyzed in corneal epithelial tissue lysates. Insilico prediction of peptides showed it to have antifungal activity which was further validated using colony forming assay and time killing kinetics. The duration of antifungal activity of peptide was improved using gelatin hydrogel through sustained delivery.ResultsVRF005 and VRF007 showed α-helical structure and was within the allowed region of Ramachandran plot. The simulation study showed their membrane penetration. The peptide uptake was found to be specific to corneal epithelial cells and also showed intracellular localization in Candida albicans and Fusarium solani. Peptides were found to be stable up to 2 hours when incubated with corneal epithelial tissue lysate. Dynamic light scattering, and native polyacrylamide gel electrophoresis revealed aggregation of peptides. VRF007 showed antifungal activity up to 24 hour whereas VRF005 showed activity up to 4 hours. Hence gelatin hydrogel-based delivery system was used to improve the activity. Actin staining of corneal epithelial cells showed that the cells were attached on gelatin hydrogel.ConclusionWe have designed corneal specific cell penetrating peptides using subtractive proteomic approach. Bioavailability and delivery of peptide was enhanced using gelatin hydrogel system.

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“…Furthermore, hPP10 fused with an RNA-binding domain could deliver small interfering RNA into cells to silence the reporter gene expression [171]. On the other hand, hPP3 (KPKRKRRKKKGHGWSR) derived from human nuclear body protein could enter cells in vitro, at a concentration-, incubation time-, serum-and temperature-dependent manner [172]. It was interesting that a CPP (TIP1) derived from toll/interleukin-1 receptor (TIR) domain-containing adapter protein suppressed toll-like receptor-mediated downstream signaling and showed therapeutic potential for TLR-mediated autoimmune and inflammatory diseases [173].…”

Section: Peptide and Protein Deliverymentioning

confidence: 99%

Cell penetrating peptides: the potent multi-cargo intracellular carriers

Kardani

Milani

Shabani

et al. 2019

Expert Opinion on Drug Delivery

152

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Introduction: Cell penetrating peptides (CPPs) known as protein translocation domains (PTD), membrane translocating sequences (MTS), or Trojan peptides (TP) are able to cross biological membranes without clear toxicity using different mechanisms, and facilitate the intracellular delivery of a variety of bioactive cargos. CPPs could overcome some limitations of drug delivery and combat resistant strains against a broad range of diseases. Despite delivery of different therapeutic molecules by CPPs, they lack cell specificity and have a short duration of action. These limitations led to design of combined cargo delivery systems and subsequently improvement of their clinical applications. Areas covered: This review covers all our studies and other researchers in different aspects of CPPs such as classification, uptake mechanisms, and biomedical applications. Expert opinion: Due to low cytotoxicity of CPPs as compared to other carriers and final degradation to amino acids, they are suitable for preclinical and clinical studies. Generally, the efficiency of CPPs was suitable to penetrate the cell membrane and deliver different cargos to specific intracellular sites. However, no CPP-based therapeutic approach has approved by FDA, yet; because there are some disadvantages for CPPs including short half-life in blood, and nonspecific CPP-mediated delivery to normal tissue. Thus, some methods were used to develop the functions of CPPs in vitro and in vivo including the augmentation of cell specificity by activatable CPPs, specific transport into cell organelles by insertion of corresponding localization sequences, incorporation of CPPs into multifunctional dendrimeric or liposomal nanocarriers to improve selectivity and efficiency especially in tumor cells.

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Highly Efficient Delivery of Functional Cargoes by a Novel Cell-Penetrating Peptide Derived from SP140-Like Protein

Cited by 28 publications

References 37 publications

Intracellular Delivery of DNA and Protein by a Novel Cell-Permeable Peptide Derived from DOT1L

Intracellular Delivery of DNA and Protein by a Novel Cell-Permeable Peptide Derived from DOT1L

Designing and enhancing the antifungal activity of corneal specific cell penetrating peptide using gelatin hydrogel delivery system

Cell penetrating peptides: the potent multi-cargo intracellular carriers

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