Methotrexate-plasmid DNA polyplexes for cancer therapy: Characterization, cancer cell targeting ability and tuned in vitro transfection

Faria, Rúben; Sousa, Ângela; Neves, Ana R.; Queiroz, João A.; Costa, Diana

doi:10.1016/j.molliq.2019.111391

Cited by 20 publications

(21 citation statements)

References 37 publications

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“…In the last decades, considerable advances have been made in the conception of nucleic acid-based delivery systems to overcome the major drawbacks of payload delivery to eukaryotic cells, namely, cellular uptake/internalization, endosomal escape, targeting a specific subcellular compartment, and ultimately, the induction of therapeutic action [19,[21][22][23][24]. In line with this aim, micelles, polymers, lipid-and peptide-based nanoparticles are among the most studied systems for gene release [25][26][27][28].…”

Section: Introductionmentioning

confidence: 99%

Development of Peptide-Based Nanoparticles for Mitochondrial Plasmid DNA Delivery

et al. 2021

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A mitochondrion is a cellular organelle able to produce cellular energy in the form of adenosine triphosphate (ATP). As in the nucleus, mitochondria contain their own genome: the mitochondrial DNA (mtDNA). This genome is particularly susceptible to mutations that are at the basis of a multitude of disorders, especially those affecting the heart, the central nervous system and muscles. Conventional clinical practice applied to mitochondrial diseases is very limited and ineffective; a clear need for innovative therapies is demonstrated. Gene therapy seems to be a promising approach. The use of mitochondrial DNA as a therapeutic, optimized by peptide-based complexes with mitochondrial targeting, can be seen as a powerful tool in the reestablishment of normal mitochondrial function. In line with this requirement, in this work and for the first time, a mitochondrial-targeting sequence (MTS) has been incorporated into previously researched peptides, to confer on them a targeting ability. These peptides were then considered to complex a plasmid DNA (pDNA) which contains the mitochondrial gene ND1 (mitochondrially encoded NADH dehydrogenase 1 protein), aiming at the formation of peptide-based nanoparticles. Currently, the ND1 plasmid is one of the most advanced bioengineered vectors for conducting research on mitochondrial gene expression. The formed complexes were characterized in terms of pDNA complexation capacity, morphology, size, surface charge and cytotoxic profile. These data revealed that the developed carriers possess suitable properties for pDNA delivery. Furthermore, in vitro studies illustrated the mitochondrial targeting ability of the novel peptide/pDNA complexes. A comparison between the different complexes revealed the most promising ones that complex pDNA and target mitochondria. This may contribute to the optimization of peptide-based non-viral systems to target mitochondria, instigating progress in mitochondrial gene therapy.

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Section: Introductionmentioning

confidence: 99%

Development of Peptide-Based Nanoparticles for Mitochondrial Plasmid DNA Delivery

et al. 2021

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“…The properties of PEI namely the molecular weight, the architecture of the chains or the degree of branching are fundamental parameters in determining the physicochemical characteristics of nucleic acids-PEI complexes. These properties also strongly influence both the cytotoxicity displayed by the complexes and the efficiency of cellular transfection [ 9 , 46 , 47 ].…”

Section: Resultsmentioning

confidence: 99%

“…Among the disadvantages of viral vectors are the immune-related toxicity with repeated administration, possible replication competence, non-targeting, or the limited insert size [ 7 , 8 ]. Cancer gene therapy has benefit from the crescent development in the design/formulation of non-viral systems for precise and targeted gene delivery [ 9 , 10 , 11 , 12 ]. Non-viral vectors can be easy and tailor-produced, can incorporate a large amount of genetic content, are biocompatible, non-toxic and non-immunogenic.…”

Section: Introductionmentioning

confidence: 99%

“…These synthetic systems have been designed to be stimuli-responsive, tunable and targeted, while promoting controlled and sustained gene release [ 13 , 14 , 15 ]. Moreover, the simultaneous release of genes and anticancer drugs to cancer cells, mediated by non-viral carriers, demonstrated to enhance the anticancer therapeutic effect [ 9 , 16 , 17 , 18 ]. In this sense, micelles, lipid, peptide and polymer-based nanoparticles have been developed to promote cellular uptake, endosomal escape, specific organelle targeting, gene delivery and, ultimately, therapeutic effect [ 19 , 20 , 21 , 22 ].…”

Section: Introductionmentioning

confidence: 99%

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Development of Tailor-Made Dendrimer Ternary Complexes for Drug/Gene Co-Delivery in Cancer

et al. 2021

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Cancer gene therapy, mediated by non-viral systems, remains a major research focus. To contribute to this field, in this work we reported on the development of dendrimer drug/gene ternary complexes. This innovative approach explored the great capacity of both polyamidoamine (PAMAM)-paclitaxel (PTX) conjugate and polyethylenimine (PEI) polymers to complex a p53-encoding plasmid DNA (pDNA), highlighting the utility of considering two compacting agents. The pDNA complexation capacity has been investigated as function of the nitrogen to phosphate groups ratio (N/P), which revealed to be a tailoring parameter. The physicochemical properties of the conceived ternary complexes were revealed and were found to be promising for cellular transfection. Furthermore, the formulated co-delivery systems demonstrated to be biocompatible. The ternary systems were able of cellular internalization and payload intracellular release. Confocal microscopy studies showed the co-localization of stained pDNA with the nucleus of cancer cells, after transfection mediated by these carriers. From this achievement, p53 gene expression occurred with the production of protein. Moreover, the activation of caspase-3 indicated apoptosis of cancer cells. This work represents a great progress on the design of dendrimer drug/gene co-delivery systems towards a more efficient cancer therapy. In this way, it instigates further in vitro studies concerning the evaluation of their therapeutic potential, expectedly supported by the synergistic effect, in tumoral cells.

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“…In comparison to conventional approaches, gene therapy allows for controlled gene release, reducing the need for multiple interventions and promotes efficient release of therapeutic genes to the tumor tissue, decreasing toxicity to healthy cells [21]. Moreover, gene therapy can be used alone or as an adjuvant to chemotherapy, since certain genes can sensitize tumor cells to drugs enhancing therapeutic outcomes by synergistic effect [22][23][24][25].…”

Section: Introductionmentioning

confidence: 99%

The Influence of Circadian Rhythm on Cancer Cells Targeting and Transfection Efficiency of a Polycation-Drug/Gene Delivery Vector

et al. 2022

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The conception of novel anticancer delivery systems and the combination of chronobiology with nanotechnology may provide a powerful tool to optimize cancer therapy. In this work, polyethylenimine (PEI) has been used to complex p53 encoded plasmid DNA (pDNA), and the anticancer drug methotrexate (MTX) has also been loaded into the vectors. To investigate the influence of circadian clock on drug/gene delivery efficiency, HeLa, C33A and fibroblast cells have been transfected with developed PEI/pDNA/MTX delivery vectors at six different time points. Phenomena as the cellular uptake/internalization, drug/gene delivery and p53 protein production have been evaluated. The cell-associated MTX fluorescence have been monitored, and p53 protein levels quantified. In HeLa and C33A cancer cells, significant levels of MTX were found for T8 and T12. For these time points, a high amount of p53 protein was quantified. Confocal microscopy images showed successful HeLa cell’s uptake of PEI/pDNA/MTX particles, at T8. In comparison, poor levels of MTX and p53 protein were found in fibroblasts; nevertheless, results indicated rhythmicity. Data demonstrate the influence of circadian rhythm on both cancer-cells targeting ability and transfection performance of PEI/pDNA/MTX carriers and seemed to provide the optimum time for drug/gene delivery. This report adds a great contribution to the field of cancer chronobiology, highlighting the relationship between circadian rhythm and nanodelivery systems, and charting the path for further research on a, yet, poorly explored but promising topic.

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Methotrexate-plasmid DNA polyplexes for cancer therapy: Characterization, cancer cell targeting ability and tuned in vitro transfection

Cited by 20 publications

References 37 publications

Development of Peptide-Based Nanoparticles for Mitochondrial Plasmid DNA Delivery

Development of Peptide-Based Nanoparticles for Mitochondrial Plasmid DNA Delivery

Development of Tailor-Made Dendrimer Ternary Complexes for Drug/Gene Co-Delivery in Cancer

The Influence of Circadian Rhythm on Cancer Cells Targeting and Transfection Efficiency of a Polycation-Drug/Gene Delivery Vector

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