The impact of microfluidic mixing of triblock micelleplexes on <i>in vitro</i> $/$ <i>in vivo</i> gene silencing and intracellular trafficking

Feldmann, Daniel P.; Xie, Yuran; Jones, Steven K.; Yu, Dongyue; Moszczyńska, Anna; Merkel, Olivia M.

doi:10.1088/1361-6528/aa6d15

Cited by 21 publications

(24 citation statements)

References 39 publications

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“…Additionally, impact on mitochondrial activity was measured in MTT assays (see Figure S4, Supplementary Material). As discussed in other work, forming nanoparticles by microfluidics leads to an increased control over the production process which in turn generates decreased hydrodynamic diameters and narrow dispersity [38,42,43]. Loy et al produced well-defined and reproducible polyplexes with controlled surface characteristics with the help of microfluidic a self-assembly based on electrostatic interaction [44].…”

Section: Discussionmentioning

confidence: 99%

“…With the previous optimization of micelleplex microfluidic mixing and in vitro/in vivo proof-ofconcept of siRNA delivery with <200 nm nanoparticles in our past study, we set out to optimize the knockdown of the proteins ERCC1 and XPF [38]. For preliminary experiments, A549 lung cancer cells were transfected with micelleplexes and Lipofectamine (LF) that contained 50 pmol of ERCC1-XPF siRNA, and protein knockdown was determined after 72 h via Western blot analysis with β-actin as a loading control.…”

Section: In Vitro Ercc1-xpf Protein Knockdownmentioning

confidence: 99%

“…Finally, the hydrophobic polymer polycaprolactone (PCL) is used to drive micelle formation and to aid in polymer cleavage due to its susceptibility toward hydrolytic degradation [37]. Previous studies performed with PEI-PCL-PEG micelleplexes demonstrated their ability to efficiently deliver siRNA in vitro to mediate sustained protein knockdown and display long-circulation profiles in vivo [38][39][40][41].…”

Section: Introductionmentioning

confidence: 99%

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Nanoparticle-Mediated Gene Silencing for Sensitization of Lung Cancer to Cisplatin Therapy

et al. 2020

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Platinum-based chemotherapy remains a mainstay treatment for the management of advanced non-small cell lung cancer. A key cellular factor that contributes to sensitivity to platinums is the 5′-3′ structure-specific endonuclease excision repair cross-complementation group 1 (ERCC1)/ xeroderma pigmentosum group F (XPF). ERCC1/XPF is critical for the repair of platinum-induced DNA damage and has been the subject of intense research efforts to identify small molecule inhibitors of its nuclease activity for the purpose of enhancing patient response to platinum-based chemotherapy. As an alternative to small molecule inhibitors, small interfering RNA (siRNA) has often been described to be more efficient in interrupting protein–protein interactions. The goal of this study was therefore to determine whether biocompatible nanoparticles consisting of an amphiphilic triblock copolymer (polyethylenimine-polycaprolactone-polyethylene glycol (PEI-PCL-PEG)) and carrying siRNA targeted to ERCC1 and XPF made by microfluidic assembly are capable of efficient gene silencing and able to sensitize lung cancer cells to cisplatin. First, we show that our PEI-PCL-PEG micelleplexes carrying ERCC1 and XPF siRNA efficiently knocked down ERCC1/XPF protein expression to the same extent as the standard siRNA transfection reagent, Lipofectamine. Second, we show that our siRNA-carrying nanoparticles enhanced platinum sensitivity in a p53 wildtype model of non-small cell lung cancer in vitro. Our results suggest that nanoparticle-mediated targeting of ERCC1/XPF is feasible and could represent a novel therapeutic strategy for targeting ERCC1/XPF in vivo.

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Section: Discussionmentioning

confidence: 99%

Section: In Vitro Ercc1-xpf Protein Knockdownmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Nanoparticle-Mediated Gene Silencing for Sensitization of Lung Cancer to Cisplatin Therapy

et al. 2020

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“…Still, further improvements are warranted and various approaches have been explored to optimize transfection efficacies of low molecular weight PEIs and/or to decrease the toxicity of high molecular weight PEIs. These include the covalent modification of PEI with polyethylene glycol (PEG) [25,26,27], the cross-linking of small PEIs with hydrolysable linkers, e.g., esters, ketals, or reductive cleavable disulfide groups leading to the reversible formation of higher molecular weights ([28,29,30,31]; see [32] for review), the copolymerization/grafting with other polymers like polycaprolactone (PCL), starch or polyvinylalcohol (PVA) [33,34,35], the modification with lipophilic molecules, e.g., fatty acids, alkanes or cholesterol ([36,37], see [38] for review), or the combination of PEI-based complexes with liposomes [39,40,41,42].…”

Section: Introductionmentioning

confidence: 99%

Polymeric Nanoparticles Based on Tyrosine-Modified, Low Molecular Weight Polyethylenimines for siRNA Delivery

et al. 2019

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A major hurdle for exploring RNA interference (RNAi) in a therapeutic setting is still the issue of in vivo delivery of small RNA molecules (siRNAs). The chemical modification of polyethylenimines (PEIs) offers a particularly attractive avenue towards the development of more efficient non-viral delivery systems. Here, we explore tyrosine-modified polyethylenimines with low or very low molecular weight (P2Y, P5Y, P10Y) for siRNA delivery. In comparison to their respective parent PEI, they reveal considerably increased knockdown efficacies and very low cytotoxicity upon tyrosine modification, as determined in different reporter and wildtype cell lines. The delivery of siRNAs targeting the anti-apoptotic oncogene survivin or the serine/threonine-protein kinase PLK1 (polo-like kinase 1; PLK-1) oncogene reveals strong inhibitory effects in vitro. In a therapeutic in vivo setting, profound anti-tumor effects in a prostate carcinoma xenograft mouse model are observed upon systemic application of complexes for survivin or PLK1 knockdown, in the absence of in vivo toxicity. We thus demonstrate the tyrosine-modification of (very) low molecular weight PEIs for generating efficient nanocarriers for siRNA delivery in vitro and in vivo, present data on their physicochemical and biological properties, and show their efficacy as siRNA therapeutic in vivo, in the absence of adverse effects.

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“…One strategy to address this limitation involves the use of microfluidic devices to increase colloidal stability by controlling the interaction of cationic polymer with anionic nucleic acid [10]. This strategy has proven useful in increasing siRNA complexation and uptake of the micelleplexes, which leads to a more efficient gene knockdown in vivo [11].…”

Section: Introductionmentioning

confidence: 99%

Microfluidic Assembly of siRNA-Loaded Micelleplexes for Tumor Targeting in an Orthotopic Model of Ovarian Cancer

Feldmann

Jones

Douglas

et al. 2019

Methods in Molecular Biology

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The use of cationic polymers to interact with negatively charged siRNA via charge complexation to form polyelectrolyte complexes has been widely studied ever since the 1998 report on RNA interference. These polyelectrolyte complex formulations aim to overcome the many pitfalls associated with the use of RNA interference as a potential cancer therapy. The triblock copolymer polyethylenimine-polycaprolactone-polyethylene glycol (PEI-PCL-PEG) contains the cation PEI and has been shown to be an efficient carrier capable of complexing with nucleic acids for gene delivery. This copolymer system also allows for targeting moieties to be linked to the micelleplex, thereby exploiting overexpressed receptors (such as the folate receptor) located within tumors. Additionally, we demonstrated recently that microfluidic mixing of PEI-PCL-PEG nanoparticles allows for the rapid, scaled-up production of micelleplexes while maintaining small and uniform particle distributions. The preparation of small and reproducible particles is imperative for clinical translation of nanomedicine and for tumor targeting via systemic administration. Furthermore, to enable tracing of its deposition in vivo after its administration, micelleplexes can be radiolabeled. To assess tumor targeting over time, the noninvasive imaging technique single-photon emission computed tomography (SPECT) offers the ability to examine the same subject at multiple time points and generate biodistribution profiles. Since the biodistribution and tumor targeting of the therapeutic load of micelleplexes is of foremost interest, we recently described an approach to modify siRNA with a DTPA (diethylenetriaminepentaacetic acid) chelator. Herein, we explain the details of encapsulating indium-labeled siRNA via microfluidic mixing in PEI-PCL-PEG nanoparticles with a folic acid targeting ligand for assessment of their in vivo tumor targeting in an orthotopic ovarian cancer model.

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The impact of microfluidic mixing of triblock micelleplexes on in vitro $/$ in vivo gene silencing and intracellular trafficking

Cited by 21 publications

References 39 publications

Nanoparticle-Mediated Gene Silencing for Sensitization of Lung Cancer to Cisplatin Therapy

Nanoparticle-Mediated Gene Silencing for Sensitization of Lung Cancer to Cisplatin Therapy

Polymeric Nanoparticles Based on Tyrosine-Modified, Low Molecular Weight Polyethylenimines for siRNA Delivery

Microfluidic Assembly of siRNA-Loaded Micelleplexes for Tumor Targeting in an Orthotopic Model of Ovarian Cancer

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