RESUMO
Demystifying the sulfation code of glycosaminoglycans (GAGs) to induce precise homing of nanoparticles in tumor cells or neurons influences the development of a potential drug- or gene-delivery system. However, GAGs, particularly heparan sulfate (HS) and chondroitin sulfate (CS), are structurally highly heterogeneous, and synthesizing well-defined HS/CS composed nanoparticles is challenging. Here, we decipher how specific sulfation patterns on HS and CS regulate receptor-mediated homing of nanoprobes in primary and secondary cells. We discovered that aggressive cancer cells such as MDA-MB-231 displayed a strong uptake of GAG-nanoprobes compared to mild or moderately aggressive cancer cells. However, there was no selectivity towards the GAG sequences, thus indicating the presence of more than one form of receptor-mediated uptake. However, U87 cells, olfactory bulb, and hippocampal primary neurons showed selective or preferential uptake of CS-E-coated nanoprobes compared to other GAG-nanoprobes. Furthermore, mechanistic studies revealed that the 4,6-O-disulfated-CS nanoprobe used the CD44 and caveolin-dependent endocytosis pathway for uptake. These results could lead to new opportunities to use GAG nanoprobes in nanomedicine.
Assuntos
Sulfatos de Condroitina , Glicosaminoglicanos , Glicosaminoglicanos/metabolismo , Heparitina Sulfato/metabolismoRESUMO
Toxoplasma gondii is a ubiquitous eukaryotic pathogen responsible for toxoplasmosis in humans and animals. This parasite is an obligate intracellular pathogen and actively invades susceptible host cells, a process which is mediated by specific receptor-ligand interactions. Here, we have identified an unnatural 2,4-disulfated d-glucuronic acid (Di-S-GlcA), a hexuronic acid composed of heparin/heparan sulfate, as a potential carbohydrate ligand that can selectively bind to T. gondii parasites. More importantly, the gelatin conjugated Di-S-GlcA multivalent probe displayed strong inhibition of parasite entry into host cells. These results open perspective for the future use of Di-S-GlcA epitopes in biomedical applications against toxoplasmosis.
Assuntos
Glucuronatos/farmacologia , Toxoplasma/efeitos dos fármacos , Adesão Celular/efeitos dos fármacos , Fibroblastos/microbiologia , Glucuronatos/síntese química , Glucuronatos/metabolismo , Humanos , Ligantes , Toxoplasma/metabolismo , Toxoplasma/patogenicidadeRESUMO
Sialic acids (Sias) are important terminal sugars on cell surfaces involved in a wide range of protein-carbohydrate interactions. Hence, agents modulating sias-mediated protein interactions are promising inhibitors or vaccine candidates. Here, we report the synthesis of Neu5Acα(2-6)Gal structural analogs and their binding to a series of siglecs. The results showed distinct binding patterns with conserved siglecs (hCD22 and mCD22) compared to rapid evolving siglecs (Siglecs -3 & -10).
RESUMO
Nanotechnology-based vaccine development necessitates understanding the crucial biophysical properties of nanostructures that alter immune responses. In this study, we demonstrate the synergistic effect of gold nanoparticles (AuNPs) shapes with toll-like receptor (TLR) agonists in immune modulation activity. Our results showed that CpG- and imidazoquinoline-conjugated rod-shaped AuNPs display relatively fast uptake by bone marrow-derived macrophage cells but exhibit poor immunogenic responses compared to their spherical and star-shaped AuNP counterparts. Surprisingly, star-shaped AuNPs exhibited intense pro-inflammatory cytokine secretion. Further mechanistic studies showed that star-shaped AuNPs were abundantly localized in the late endosome and lysosomal regions, whereas rod-shaped AuNPs were majorly sequestered in the mitochondrial region. These findings reveal that the shape of the nanostructures plays a pivotal role in driving the adjuvant molecules toward their receptors and altering immune responses.
Assuntos
Ouro , Nanopartículas Metálicas , Ouro/química , Nanopartículas Metálicas/uso terapêutico , Adjuvantes Imunológicos/farmacologia , Macrófagos , ImunidadeRESUMO
Nanoparticles (NPs) embedded with bioactive ligands such as carbohydrates, peptides, and nucleic acid have emerged as a potential tool to target biological processes. Traditional inâ vitro assays performed under statistic conditions may result in non-specific outcome sometimes, mainly because of the sedimentation and self-assembly nature of NPs. Inverted cell-culture assay allows for flexible and accurate detection of the receptor-mediated uptake and cytotoxicity of NPs. By combining this technique with glyco-gold nanoparticles, cellular internalization and cytotoxicity were investigated. Regioselective glycosylation patterns and shapes of the NPs could tune the receptors' binding affinity, resulting in precise cellular uptake of gold nanoparticles (AuNPs). Two cell lines HepG2 and HeLa were probed with galactosamine-embedded fluorescent AuNPs, revealing significant differences in cytotoxicity and uptake mechanism in upright and invert inâ vitro cell-culture assay, high-specificity toward uptake, and allowing for a rapid screening and optimization technique.
Assuntos
Antineoplásicos/farmacologia , Técnicas de Cultura de Células , Galactosamina/farmacologia , Ouro/farmacologia , Nanopartículas/química , Antineoplásicos/química , Proliferação de Células/efeitos dos fármacos , Ensaios de Seleção de Medicamentos Antitumorais , Galactosamina/química , Ouro/química , Células HeLa , Células Hep G2 , Humanos , Estrutura Molecular , Tamanho da Partícula , Células Tumorais CultivadasRESUMO
Sialic acid-conjugated nanocarriers have emerged as attractive biomarkers with promising biomedical applications. The translation of these nanocarriers into clinical applications requires in-depth assessment in animal models. However, due to the complexity, ethical concerns, and cost of the high-order animal system, there is an immediate need of information-rich simple animal models to decipher the biological significance. Herein, we performed in vivo head-to-head comparison of Neu5Acα(2-6) and α(2-3)Gal conjugated quantum dots (QDs) toxicity, biodistribution, and sequestration in wild-type zebrafish ( Danio rerio) and mouse model (C57BL). The fluorescent properties and cadmium composition of quantum dots were used to map the blood clearance, biodistribution, and sequestration of the sialylated QDs in major organs of both models. We observed that α(2-6) sialylated QDs preferentially have prolonged circulating half-life and broader biodistribution in both models. On the contrary, α(2-3) sialic acid and galactose-conjugated QDs have shortened blood circulation time and are sequestered in the liver, and cleared after several hours in both models. These results demonstrate the applicability of the zebrafish and sialylated QDs to target specific organs, as well as drug delivery and biomedical diagnostics.
Assuntos
Pontos Quânticos , Animais , Camundongos , Camundongos Endogâmicos C57BL , Ácido N-Acetilneuramínico , Distribuição Tecidual , Peixe-ZebraRESUMO
Understanding blood group antigen binding preferences for C-type lectin receptors holds promise for modulating immune responses, since several Gram-negative bacteria express blood group antigens as molecular mimicry to evade immune responses. Herein, we report the synthesis of ABO blood group antigen active tri and disaccharides to investigate the binding specificity with various C-type lectin receptors using glycan microarray. The results of binding preferences show that distinct glycosylation on the galactose and fucose motifs are key for C-type lectin receptor binding and that these interactions occur in a Ca2+-dependent fashion.
Assuntos
Sistema ABO de Grupos Sanguíneos/imunologia , Sistema ABO de Grupos Sanguíneos/metabolismo , Lectinas Tipo C/metabolismo , Polissacarídeos/imunologia , Animais , Dissacarídeos/química , Dissacarídeos/imunologia , Glicosilação , Humanos , Camundongos , Estrutura Molecular , Polissacarídeos/química , Análise Serial de Proteínas , Ligação Proteica/imunologiaRESUMO
Glyconanotechnology offers a broad range of applications across basic and translation research. Despite the tremendous progress in glyco-nanomaterials, there is still a huge gap between the basic research and therapeutic applications of these molecules. It has been reported that complexity and the synthetic challenges in glycans synthesis, the cost of the high order in vivo models and large amount of sample consumptions limited the effort to translate the glyco-nanomaterials into clinical applications. In this regards, several promising simple animal models for preliminary, quick analysis of the nanomaterials activities has been proposed. Herein, we have studied a systematic evaluation of the toxicity, biodistribution of fluorescently tagged PEG and mannose-capped gold nanoparticles (AuNPs) of three different shapes (sphere, rod, and star) in the adult zebrafish model, which could accelerate and provide preliminary results for further experiments in the higher order animal system. ICP-MS analysis and confocal images of various zebrafish organs revealed that rod-AuNPs exhibited the fast uptake. While, star-AuNPs displayed prolong sequestration, demonstrating its potential therapeutic efficacy in drug delivery.