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喷雾干燥的脂质纳米颗粒配方使mRNA的内内传递成为可能
Kristina Pagh Friis1, Sandra Gracin2, Steven Oag3
1Advanced Drug Delivery, Pharmaceutical Sciences, R&D, AstraZeneca, Gothenburg, Sweden.
概括
喷雾干燥可以提高脂质纳米粒子 (LNP) mRNA的稳定性,从而提高潜在的肺部输送. 这种新的配方克服了冷链储存的挑战,使得新的基于RNA的疗法,如疫苗和蛋白质替代疗法.
科学领域:
- 生物技术是生物技术.
- 纳米医学是一种纳米医学.
- 药物运输 药物运输 药物运输
背景情况:
- 脂质纳米粒子 (LNP) 输送系统对于RNA疗法至关重要,包括siRNA和mRNA疫苗.
- 储存和分发期间的稳定性,特别是没有冷链,仍然是非病毒RNA药物输送的重大挑战.
研究的目的:
- 为了设计一个稳定的,喷雾干燥的脂质纳米粒子 (LNP) mRNA配方.
- 与液体配方相比,评估喷雾干燥的LNP mRNA的稳定性和功能.
- 评估用于肺基RNA治疗的喷雾干燥LNP的内传递潜力.
主要方法:
- 为LNP mRNA配方开发喷雾干燥工艺.
- 液体与喷雾干燥LNPmRNA配方在受控储存条件下 (4°C两周) 的比较稳定性评估.
- 喷雾干燥的LNP含有增强绿色光蛋白 (eGFP) mRNA的内注射给大鼠.
- 在分娩后对各种肺细胞类型的EGFP蛋白表达的分析.
主要成果:
- 喷雾干燥产生了一个稳定的干粉LNP mRNA配方.
- 喷雾干燥配方保持了mRNA功能,并且在储存后与液体配方相比表现有所改善.
- 喷雾干燥的LNP的内内输送成功导致了肺细胞 (包括上皮细胞和巨细胞) 中的eGFP蛋白表达.
结论:
- 喷雾干燥显著提高了LNP mRNA配方的稳定性,解决了RNA药物分发中的一个关键挑战.
- 这种方法有望通过吸入使RNA传递到肺部.
- 潜在的应用包括蛋白质替代疗法,基因编辑,以及针对呼吸道病原体的吸入疫苗的开发.
相关概念视频
MicroRNAs
MicroRNA (miRNA) are short, regulatory RNA transcribed from introns—non-coding regions of a gene—or intergenic regions—stretches of DNA present between genes. Several processing steps are required to form biologically active, mature miRNA. The initial transcript, called primary miRNA (pri-mRNA), base-pairs with itself forming a stem-loop structure. Within the nucleus, an endonuclease enzyme, called Drosha, shortens the stem-loop structure into hairpin-shaped pre-miRNA. After the pre-miRNA ends...
Nonsense-mediated mRNA Decay
The Upf proteins that carry out nonsense-mediated decay (NMD) are found in all eukaryotic organisms, including humans. Each protein has an individual role, but they need to work in collaboration. Upf1 is an ATP-dependent RNA helicase that unwinds the RNA helix. Because Upf1 can unwind any RNA, Upf2 and Upf3 are required to help Upf1 discriminate between nonsense and normal mRNAs.
Usually, Upf3 binds to an Exon Junction Complex (EJC) at mRNA splice sites. If a ribosome fully translates the mRNA,...
Usually, Upf3 binds to an Exon Junction Complex (EJC) at mRNA splice sites. If a ribosome fully translates the mRNA,...
mRNA Stability and Gene Expression
The structure and stability of mRNA molecules regulates gene expression, as mRNAs are a key step in the pathway from gene to protein. In eukaryotes, the half-life of mRNA varies from a few minutes up to several days. mRNA stability is essential in growth and development. The absence of the proteins regulating its stability, such as tristetraprolin in mice, can cause systemic issues, including bone marrow overgrowth, inflammation, and autoimmunity.
Cis-acting Elements involved in mRNA stability
Cis-acting Elements involved in mRNA stability
Nonsense-mediated mRNA Decay
The Upf proteins that carry out nonsense-mediated decay (NMD) are found in all eukaryotic organisms, including humans. Each protein has an individual role, but they need to work in collaboration. Upf1 is an ATP-dependent RNA helicase that unwinds the RNA helix. Because Upf1 can unwind any RNA, Upf2 and Upf3 are required to help Upf1 discriminate between nonsense and normal mRNAs.
Usually, Upf3 binds to an Exon Junction Complex (EJC) at mRNA splice sites. If a ribosome fully translates the mRNA,...
Usually, Upf3 binds to an Exon Junction Complex (EJC) at mRNA splice sites. If a ribosome fully translates the mRNA,...
MicroRNAs
MicroRNA (miRNA) are short, regulatory RNA transcribed from introns (non-coding regions of a gene) or intergenic regions (stretches of DNA present between genes). Several processing steps are required to form biologically active, mature miRNA. The initial transcript, called primary miRNA (pri-mRNA), base-pairs with itself, forming a stem-loop structure. Within the nucleus, an endonuclease enzyme, called Drosha, shortens the stem-loop structure into hairpin-shaped pre-miRNA. After the pre-miRNA...
Nucleic Acid Structure
The pentose sugar in DNA is deoxyribose, while in RNA the pentose sugar is ribose. The difference between the sugars is the presence of the hydroxyl group on the ribose's second carbon and a hydrogen on the deoxyribose's second carbon. The phosphate residue attaches to the hydroxyl group of the 5′ carbon of one sugar and the hydroxyl group of the 3′ carbon of the sugar of the next nucleotide, which forms a 5′ to 3′ phosphodiester linkage.
DNA Structure
DNA has a double-helix structure. The...
DNA Structure
DNA has a double-helix structure. The...

