通过在单个纳米载体上共同递送信使 RNA 和质粒 DNA,实现核酸编码蛋白质的时间交错表达。

IF 5.7 3区 医学 Q1 MEDICINE, RESEARCH & EXPERIMENTAL
Drug Delivery and Translational Research Pub Date : 2024-12-01 Epub Date: 2024-07-15 DOI:10.1007/s13346-024-01668-w
Sarah S Nasr, Pascal Paul, Brigitta Loretz, Claus-Michael Lehr
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引用次数: 0

摘要

在未来几年里,基于核酸(NA)的联合疗法领域中的一个突出要求是同时递送不同的蛋白质编码多核苷酸种类,而这些多核苷酸种类的治疗产物的表达动力学各不相同。目前的研究探讨了通过在同一纳米载体的内核中同时递送质粒 DNA(pDNA)和外壳上的 mRNA 来实现编码蛋白质的时间交错表达的能力。内核以 A 型明胶-pDNA 共蒸物为基础,经过热稳定处理后形成不可逆的纳米凝胶,其稳定性足以沉积阳离子涂层(即硫酸原胺或 LNP 相关脂质混合物)。只有原胺包被的纳米载体在装载 mRNA 后仍保持胶体稳定,并能成功地将荧光报告基因 mRNA(mCherry)和 pDNA(pAmCyan1)共转染小鼠树突状细胞系 DC2.4。仅对原胺包被的纳米系统进行了进一步研究,比较了同一纳米载体同时转染 mRNA 和 pDNA 的转染效率(转染细胞百分比)和蛋白表达水平(平均荧光强度,MFI),并使用流式细胞仪对 DC2.4 在 48 小时内的转染情况进行了动力学评估。两种核苷酸的转染起始时间都有所延迟,转染水平为
本文章由计算机程序翻译,如有差异,请以英文原文为准。

Realizing time-staggered expression of nucleic acid-encoded proteins by co-delivery of messenger RNA and plasmid DNA on a single nanocarrier.

Realizing time-staggered expression of nucleic acid-encoded proteins by co-delivery of messenger RNA and plasmid DNA on a single nanocarrier.

Co-delivery of different protein-encoding polynucleotide species with varying expression kinetics of their therapeutic product will become a prominent requirement in the realm of combined nucleic acid(NA)-based therapies in the upcoming years. The current study explores the capacity for time-staggered expression of encoded proteins by simultaneous delivery of plasmid DNA (pDNA) in the core and mRNA on the shell of the same nanocarrier. The core is based on a Gelatin Type A-pDNA coacervate, thermally stabilized to form an irreversible nanogel stable enough for the deposition of cationic coats namely, protamine sulfate or LNP-related lipid mixtures. Only the protamine-coated nanocarriers remained colloidally stable following mRNA loading and could successfully co-transfect murine dendritic cell line DC2.4 with fluorescent reporter mRNA(mCherry) and pDNA (pAmCyan1). Further investigation of the protamine-coated nanosystem only, the transfection efficiency (percentage of transfected cells) and level of protein expression (mean fluorescence intensity, MFI) of mRNA and pDNA, simultaneously delivered by the same nanocarrier, were compared and kinetically assessed over 48 h in DC2.4 using flow cytometry. The onset of transfection for both nucleotides was initially delayed, with levels < 5% at 6 h. Thereafter, mRNA transfection reached 90% after 24 h and continued to slightly increase until 48 h. In contrast, pDNA transfection was clearly slower, reaching approximately 40% after 24 h, but continuing to increase to reach 94% at 48 h. The time course of protein expression (represented by MFI) for both NAs essentially followed that of transfection. Model-independent as well as model-dependent kinetic parameters applied to the data further confirmed such time-staggered expression of the two NA's where mRNA's rate of transfection and protein expression initially exceeded those of pDNA in the first 24 h of the experiment whereas the opposite was true during the second 24 h of the experiment where pDNA displayed the higher response rates. We expect that innovative nanocarriers capable of time-staggered co-delivery of different nucleotides could open new perspectives for multi-dosing, pulsatile or sustained expression of nucleic acid-based therapeutics in protein replacement, vaccination, and CRISPR-mediated gene editing scenarios.

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来源期刊
Drug Delivery and Translational Research
Drug Delivery and Translational Research MEDICINE, RESEARCH & EXPERIMENTALPHARMACOL-PHARMACOLOGY & PHARMACY
CiteScore
11.70
自引率
1.90%
发文量
160
期刊介绍: The journal provides a unique forum for scientific publication of high-quality research that is exclusively focused on translational aspects of drug delivery. Rationally developed, effective delivery systems can potentially affect clinical outcome in different disease conditions. Research focused on the following areas of translational drug delivery research will be considered for publication in the journal. Designing and developing novel drug delivery systems, with a focus on their application to disease conditions; Preclinical and clinical data related to drug delivery systems; Drug distribution, pharmacokinetics, clearance, with drug delivery systems as compared to traditional dosing to demonstrate beneficial outcomes Short-term and long-term biocompatibility of drug delivery systems, host response; Biomaterials with growth factors for stem-cell differentiation in regenerative medicine and tissue engineering; Image-guided drug therapy, Nanomedicine; Devices for drug delivery and drug/device combination products. In addition to original full-length papers, communications, and reviews, the journal includes editorials, reports of future meetings, research highlights, and announcements pertaining to the activities of the Controlled Release Society.
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