Cancer gene therapy mediated by RALA/plasmid DNA vectors: Nitrogen to phosphate groups ratio (N/P) as a tool for tunable transfection efficiency and apoptosis

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

doi:10.1016/j.colsurfb.2019.110610

Cited by 29 publications

(25 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]. In particular, cell-penetrating peptides (CPP) offer exceptional properties to be explored as gene delivery vehicles for successful gene therapy [28][29][30][31][32].…”

Section: Introductionmentioning

confidence: 99%

“…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]. In particular, cell-penetrating peptides (CPP) offer exceptional properties to be explored as gene delivery vehicles for successful gene therapy [28][29][30][31][32]. CPPs are short peptides, usually of fewer than 30 amino acids, and can be separated into arginine-rich and amphipathic peptides.…”

Section: Introductionmentioning

confidence: 99%

“…CPPs are short peptides, usually of fewer than 30 amino acids, and can be separated into arginine-rich and amphipathic peptides. The amphipathic ones possess both hydrophilic and hydrophobic domains that confer on these peptides the ability to interact with the genetic material, promoting its membrane translocation, followed by cell entry [28,30,33]. However, the exact mechanism CPPs use to penetrate cells is still not fully understood, with endocytosis and direct penetration being the most probable hypothesis [30,33].…”

Section: Introductionmentioning

confidence: 99%

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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%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Development of Peptide-Based Nanoparticles for Mitochondrial Plasmid DNA Delivery

et al. 2021

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“…Moreover, Neves et al have reached to a decrease in HeLa cell viability by 41% at 48 h after transfection with RALA/ p53-encoding pDNA nanoparticles [27]. The fact that these studies involved the use of nanoparticles for targeted delivery of cancer cells, perhaps leads to an increase in transfection efficiency and p53 expression, affecting the cellular viability in a more pronounced way.…”

Section: Discussionmentioning

confidence: 99%

The Performance of Minicircle DNA Versus Parental Plasmid in p53 Gene Delivery Into HPV-18-Infected Cervical Cancer Cells

Eusébio

Almeida

Alves

et al. 2021

Nucleic Acid Therapeutics

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Minicircle DNA (mcDNA) has been suggested as a vanguard technology for gene therapy, consisting of a nonviral DNA vector devoid of prokaryotic sequences. Unlike conventional plasmid DNA (pDNA), this small vector is able to sustain high expression rates throughout time. Thus, this work describes the construction, production, and purification of mcDNA-p53 and its precursor parental plasmid (PP)-p53 for a comparative study of both DNA vectors in the growth suppression of human papillomavirus (HPV)-18-infected cervical cancer cells. First, live cell imaging and fluorescence microscopy studies allowed to understand that mcDNA-p53 vector was able to enter cell nuclei more rapidly than PP-p53 vector, leading to a transfection efficiency of 68% against 34%, respectively. Then, p53 transcripts and protein expression assessment revealed that both vectors were able to induce transcription and the target protein expression. However, the mcDNA-p53 vector performance stood out, by demonstrating higher p53 expression levels (91.65-2.82 U/mL vs. 74.75-4.44 U/mL). After assuring the safety of both vectors by viability studies, such potential was confirmed by proliferation and apoptosis assays. These studies confirmed the mcDNA-p53 vector function toward cell cycle arrest and apoptosis in HPV-18infected cervical cancer cells. Altogether, these results suggest that the mcDNA vector has a more promising and efficient role as a DNA vector than conventional pDNA, opening new investigation lines for cervical cancer treatment in the future.

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“…have characterized the peptide RALA/p53 encoding pDNA vectors at different nitrogen to phosphate groups (N/P) ratios. They found that, in comparison with N/P as 2, systems with N/P from 5 to 10 had smaller particle size, more positively charged surfaces, and higher pDNA encapsulation capacity ( 72 ), which in turn led to better cell internalization, higher p53 protein expression levels, and large extent of apoptosis when tested using HeLa cells. Sainz-Ramos et al .…”

Section: Effects Of Gene Vector Properties and Formulation Factors On In Vitro And In Vivo Performamentioning

confidence: 99%

Non-viral Vectors in Gene Therapy: Recent Development, Challenges, and Prospects

Gao

2021

AAPS J

240

161

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Gene therapy has been experiencing a breakthrough in recent years, targeting various specific cell groups in numerous therapeutic areas. However, most recent clinical studies maintain the use of traditional viral vector systems, which are challenging to manufacture cost-effectively at a commercial scale. Non-viral vectors have been a fast-paced research topic in gene delivery, such as polymers, lipids, inorganic particles, and combinations of different types. Although non-viral vectors are low in their cytotoxicity, immunogenicity, and mutagenesis, attracting more and more researchers to explore the promising delivery system, they do not carry ideal characteristics and have faced critical challenges, including gene transfer efficiency, specificity, gene expression duration, and safety. This review covers the recent advancement in non-viral vectors research and formulation aspects, the challenges, and future perspectives.

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Cancer gene therapy mediated by RALA/plasmid DNA vectors: Nitrogen to phosphate groups ratio (N/P) as a tool for tunable transfection efficiency and apoptosis

Cited by 29 publications

References 37 publications

Development of Peptide-Based Nanoparticles for Mitochondrial Plasmid DNA Delivery

Development of Peptide-Based Nanoparticles for Mitochondrial Plasmid DNA Delivery

The Performance of Minicircle DNA Versus Parental Plasmid in p53 Gene Delivery Into HPV-18-Infected Cervical Cancer Cells

Non-viral Vectors in Gene Therapy: Recent Development, Challenges, and Prospects

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