Size-Controlled and Shelf-Stable DNA Particles for Production of Lentiviral Vectors

Hu, Yizong; Zhu, Yining; Sutherland, Nolan; Wilson, David R.; Pang, Marion; Liu, Ester; Staub, Jacob R.; Berlinicke, Cynthia; Zack, Donald J.; Green, Jordan J.; Reddy, Sashank K.; Mao, Hai‐Quan

doi:10.1021/acs.nanolett.1c01421

Cited by 22 publications

(26 citation statements)

References 32 publications

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“…Chemical approaches to enhancing the uptake of DNA-based vaccines do not require a secondary physical application, but rather utilize delivery cargos such as chitosan and polyethylenimine (PEI) (Kudsiova et al, 2008;Li et al, 2012). Such delivery cargos have a size range of 10-1,000 nm, which falls within the size range of the materials employed in this study (Khanmohammadi et al, 2015;Hu et al, 2021). Prediction of the lateral distribution of such delivery cargos should follow the power law distribution presented in Table 2.…”

Section: Discussionmentioning

confidence: 95%

Molecular distribution in intradermal injection for transfer and delivery of therapeutics

et al. 2023

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Intradermal (ID) injection is a technique widely used in laboratorial and clinical applications. The boundary of the dome-like bleb formed during injection is assumed to represent the lateral extent of the injected material. This work systematically characterizes cargo molecule distribution (puddle) as a function of injection volume and molecular/particle size in rat skin post ID injection. In general, results indicate that the puddle forms a subdomain laterally contained within the bleb, with an area inversely correlating to the molecular size of the injected material. For 50 μL and 100 µL injections, the average area of the bleb was 40.97 ± 6.30 mm2 and 55.64 ± 8.20 mm2, respectively, regardless of the molecular/particle size. On the other hand, the area of the puddle was dependent on the molecular size and ranged between 45.38 ± 8.29 mm2 and 6.14 ± 4.50 mm2 for 50 µL injections, and 66.64 ± 11.22 mm2 and 11.50 ± 9.67 mm2 for 100 µL injections. The lateral distribution appears to have no time-dependency up to 10 min post injection. The trend in the depth of cargo penetration is also similar, with smaller particles extending deeper into the dermis and subcutaneous fat layers. Because the area of puddle can be significantly less than that of the bleb, establishing base characterization is essential to understand cellular interactions with the injected biological substances.

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Section: Discussionmentioning

confidence: 95%

Molecular distribution in intradermal injection for transfer and delivery of therapeutics

et al. 2023

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“…6 f and g), whereas no significant difference was observed in the transfection efficiencies of DI-6 and FIII-1 compared to the drastic increases for the top four formulations. Furthermore, the endosomal escape capability of selected formulations was examined by quantitative Cellomics high-content analysis on an established B16-Gal8-GFP cell line 33 , 34 . The results showed in Fig.…”

Section: Resultsmentioning

confidence: 99%

Multi-step screening of DNA/lipid nanoparticles and co-delivery with siRNA to enhance and prolong gene expression

et al. 2022

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Lipid nanoparticles hold great potential as an effective non-viral vector for nucleic acid-based gene therapy. Plasmid DNA delivery can result in extended transgene expression compared to mRNA-based technologies, yet there is a lack of systematic investigation into lipid nanoparticle compositions for plasmid DNA delivery. Here, we report a multi-step screening platform to identify optimized plasmid DNA lipid nanoparticles for liver-targeted transgene expression. To achieve this, we analyze the role of different helper lipids and component ratios in plasmid DNA lipid nanoparticle-mediated gene delivery in vitro and in vivo. Compared to mRNA LNPs and in vivo-jetPEI/DNA nanoparticles, the identified plasmid DNA lipid nanoparticles successfully deliver transgenes and mediate prolonged expression in the liver following intravenous administration in mice. By addressing different physiological barriers in a stepwise manner, this screening platform can efficiently down select effective lipid nanoparticle candidates from a lipid nanoparticle library of over 1000 formulations. In addition, we substantially extend the duration of plasmid DNA nanoparticle-mediated transgene expression using a DNA/siRNA co-delivery approach that targets transcription factors regulating inflammatory response pathways. This lipid nanoparticle-based co-delivery strategy further highlights the unique advantages of an extended transgene expression profile using plasmid DNA delivery and offers new opportunities for DNA-based gene medicine applications.

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“…The results are presented in Figure 2 . The pDNA-based complexes were formed by a combination of electrostatic and hydrophobic interactions with the consequent formation of nanoparticles [ 50 , 55 , 56 ]. The arginine residues of WRAP5 mainly contributed to the electrostatic interaction with pDNA while tryptophan residues, most probably, interacted with the minor groove of pDNA by hydrophobic interactions [ 21 ].…”

Section: Resultsmentioning

confidence: 99%

Development of WRAP5 Peptide Complexes for Targeted Drug/Gene Co-Delivery toward Glioblastoma Therapy

et al. 2022

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Despite the great progress over the past few decades in both the diagnosis and treatment of a great variety of human cancers, glioblastoma remains the most lethal brain tumor. In recent years, cancer gene therapy focused on non-viral vectors which emerged as a promising approach to glioblastoma treatment. Transferrin (Tf) easily penetrates brain cells of the blood–brain barrier, and its receptor is highly expressed in this barrier and glioblastoma cells. Therefore, the development of delivery systems containing Tf appears as a reliable strategy to improve their brain cells targeting ability and cellular uptake. In this work, a cell-penetrating peptide (WRAP5), bearing a Tf-targeting sequence, has been exploited to condense tumor suppressor p53-encoding plasmid DNA (pDNA) for the development of nanocomplexes. To increase the functionality of developed nanocomplexes, the drug Temozolomide (TMZ) was also incorporated into the formulations. The physicochemical properties of peptide/pDNA complexes were revealed to be dependent on the nitrogen to phosphate groups ratio and can be optimized to promote efficient cellular internalization. A confocal microscopy study showed the capacity of developed complexes for efficient glioblastoma cell transfection and consequent pDNA delivery into the nucleus, where efficient gene expression took place, followed by p53 protein production. Of promise, these peptide/pDNA complexes induced a significant decrease in the viability of glioblastoma cells. The set of data reported significantly support further in vitro research to evaluate the therapeutic potential of developed complexes against glioblastoma.

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Size-Controlled and Shelf-Stable DNA Particles for Production of Lentiviral Vectors

Cited by 22 publications

References 32 publications

Molecular distribution in intradermal injection for transfer and delivery of therapeutics

Molecular distribution in intradermal injection for transfer and delivery of therapeutics

Multi-step screening of DNA/lipid nanoparticles and co-delivery with siRNA to enhance and prolong gene expression

Development of WRAP5 Peptide Complexes for Targeted Drug/Gene Co-Delivery toward Glioblastoma Therapy

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