Navegando por Palavras-chave "Cell penetrating peptide"
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- ItemAcesso aberto (Open Access)Aggregation limiting cell-penetrating peptides derived from protein signal sequences(MDPI, 2023-02-16) da Silva, Emerson Rodrigo [UNIFESP]; Bicev, Renata Naporano [UNIFESP]; Porosk, Ly; Härk, Heleri; Armolik, Eger-Jasper; Langel, Ülo; Nebogatova, Jekaterina; Gaidutsik, Ilja; Arukuust, Piret; http://lattes.cnpq.br/7800589206457326Alzheimer’s disease (AD) is the most common neurodegenerative disease (ND) and the leading cause of dementia. It is characterized by non-linear, genetic-driven pathophysiological dynamics with high heterogeneity in the biological alterations and the causes of the disease. One of the hallmarks of the AD is the progression of plaques of aggregated amyloid-β (Aβ) or neurofibrillary tangles of Tau. Currently there is no efficient treatment for the AD. Nevertheless, several breakthroughs in revealing the mechanisms behind progression of the AD have led to the discovery of possible therapeutic targets. Some of these include the reduction in inflammation in the brain, and, although highly debated, limiting of the aggregation of the Aβ. In this work we show that similarly to the Neural cell adhesion molecule 1 (NCAM1) signal sequence, other Aβ interacting protein sequences, especially derived from Transthyretin, can be used successfully to reduce or target the amyloid aggregation/aggregates in vitro. The modified signal peptides with cell-penetrating properties reduce the Aβ aggregation and are predicted to have anti-inflammatory properties. Furthermore, we show that by expressing the Aβ-EGFP fusion protein, we can efficiently assess the potential for reduction in aggregation, and the CPP properties of peptides in mammalian cells.
- ItemAcesso aberto (Open Access)Amyloid-like self-assembly of a hydrophobic cell-penetrating peptide and its use as a carrier for nucleic acids(American Chemical Society, 2021-08-04) da Silva, Emerson Rodrigo [UNIFESP]; Mello, Lucas Rodrigues de [UNIFESP]; Porosk, Ly; Lourenço, Thiago da Costa [UNIFESP]; Garcia, Bianca Bonetto Moreno [UNIFESP]; Costa, Carlos Alberto Rodrigues; Han, Sang Won [UNIFESP]; Souza, Juliana dos Santos de; Langel, Ülo; http://lattes.cnpq.br/7800589206457326; http://lattes.cnpq.br/4209242593570615; http://lattes.cnpq.br/4726216622634724; http://lattes.cnpq.br/7341563736516694; http://lattes.cnpq.br/2833950143696339; http://lattes.cnpq.br/0069955147703693; http://lattes.cnpq.br/7929949468269206Cell-penetrating peptides (CPPs) are a topic subject potentially exploitable for creating new nanotherapeutics for the delivery of bioactive loads. These compounds are often classified into three major categories according to their physicochemical characteristics: cationic, amphiphilic, and hydrophobic. Among them, the group of hydrophobic CPPs has received increasing attention in recent years due to toxicity concerns posed by highly cationic CPPs. The hexapeptide PFVYLI (P: proline, F: phenylalanine, V: valine, Y: tyrosine, L: leucine and I: isoleucine), a fragment derived from the C-terminal portion of α1-antitrypsin, is a prototypal example of hydrophobic CPP. This sequence shows reduced cytotoxicity, capacity of nuclear localization, and its small size readily hints suitability as a building block to construct nanostructured materials. In this study, we examine the self-assembling properties of PFVYLI and investigate its ability to form non-covalent complexes with nucleic acids. By using a combination of biophysical tools including synchrotron small-angle X-ray scattering and atomic force microscopy-based infrared spectroscopy, we discovered that this CPP self-assembles into discrete nanofibrils with remarkable amyloidogenic features. Over the course of days, these fibrils coalesce into rod-like crystals that easily reach the micrometer range. Despite lacking cationic residues in the composition, PFVYLI forms non-covalent complexes with nucleic acids that retain -sheet pairing found in amyloid aggregates. In vitro vectorization experiments performed with double-stranded DNA fragments indicate that complexes promote the internalization of nucleic acids, revealing that tropism toward cell membranes is preserved upon complexation. On the other hand, transfection assays with splice-correction oligonucleotides (SCOs) for luciferase expression show limited bioactivity across a narrow concentration window, suggesting that propensity to form amyloidogenic aggregates may trigger endosomal entrapment. We anticipate that the findings presented here open perspectives for using this archetypical hydrophobic CPP in the fabrication of nanostructured scaffolds, which potentially integrate properties of amyloids and translocation capabilities of CPPs.
- ItemSomente MetadadadosDesign and characterization of crotamine-functionalized gold nanoparticles(Elsevier Science Bv, 2018) Karpel, Richard L.; Liberato, Michelle da Silva; Campeiro, Joana Darc [UNIFESP]; Bergeon, Lorna; Szychowski, Brian; Butler, Andrew; Marino, Giovanni; Cusic, Joelle F.; Oliveira, Lilian Caroline Goncalves de [UNIFESP]; Oliveira, Eduardo B.; Farias, Marcelo Alexandre de; Portugal, Rodrigo Villares; Alves, Wendel Andrade; Daniel, Marie-Christine; Hayashi, Mirian Akemi Furuie [UNIFESP]This paper describes the development of a facile and environmentally friendly strategy for supporting crotamine on gold nanoparticles (GNPs). Our approach was based on the covalent binding interaction between the cell penetrating peptide crotamine, which is a snake venom polypeptide with preference to penetrate dividing cells, and a polyethylene glycol (PEG) ligand, which is a nontoxic, water-soluble and easily obtainable commercial polymer. Crotamine was derivatized with ortho-pyridyldisulfide-polyethyleneglycol-N-hydroxysuccinimide (OPSS-PEG-SVA) cross-linker to produce OPSS-PEG-crotamine as the surface modifier of GNP. OPSS-PEG-SVA can serve not only as a surface modifier, but also as a stabilizing agent for GNPs. The successful PEGylation of the nanoparticles was demonstrated using different physicochemical techniques, while the grafting densities of the PEG ligands and crotamine on the surface of the nanoparticles were estimated using a combination of electron microscopy and mass spectrometry analysis. In vitro assays confirmed the internalization of these GNPs, into living HeLa cells. The results described herein suggest that our approach may serve as a simple platform for the synthesis Of GNPs decorated with crotamine with well-defined morphologies and uniform dispersion, opening new roads for crotamine biomedical applications. (C) 2017 Elsevier B.V. All rights reserved.
- ItemAcesso aberto (Open Access)Modification of the linker amino acid in the cell-penetrating peptide NickFect55 leads to enhanced pDNA transfection for in vivo applications(MDPI, 2023-02-20) Porosk, Ly; Mello, Lucas; da Silva, Emerson Rodrigo [UNIFESP]; Härk, Heleri; Kurrikoff, Kaido; Arukuust, Piret; http://lattes.cnpq.br/7800589206457326Despite numerous efforts over the last three decades, nucleic acid-based therapeutics still lack delivery platforms in the clinical stage. Cell-penetrating peptides (CPPs) may offer solutions as potential delivery vectors. We have previously shown that designing a “kinked” structure in the peptide backbone resulted in a CPP with efficient in vitro transfection properties. Further optimization of the charge distribution in the C-terminal part of the peptide led to potent in vivo activity with the resultant CPP NickFect55 (NF55). Currently, the impact of the linker amino acid was further investigated in the CPP NF55, with the aim to discover potential transfection reagents for in vivo application. Taking into account the expression of the delivered reporter in the lung tissue of mice, and the cell transfection in the human lung adenocarcinoma cell line, the new peptides NF55-Dap and NF55-Dab* have a high potential for delivering nucleic acid-based therapeutics to treat lung associated diseases, such as adenocarcinoma.