从冷保护剂和缓冲查中对冷化mRNA-LNP的物理化学和结构见解
Yuchen Fan1, Diamanda Rigas2, Lee Joon Kim3
1Synthetic Molecule Pharmaceutical Sciences, Genentech Inc., 1 DNA Way, South San Francisco, CA 94080, USA.
概括
淋化可以稳定信使RNA-脂质纳米粒子 (mRNA-LNP) 疗法,消除超冷储存需求. 新型辅助剂,如聚烯利 (PVP),有望提高mRNA-LNP的稳定性和功能.
科学领域:
- 制药科学 制药科学
- 生物技术是生物技术.
- 材料科学 材料科学 材料科学
背景情况:
- 使者RNA (mRNA) 疗法,包括脂质纳米粒子 (LNP) 配方,已经显示出显著的希望,但需要超冷储存.
- 这种超冷链物流对mRNA-LNP疗法的全球分销和可访问性提出了重大挑战.
- 软化提供了一个潜在的解决方案,以提高mRNA-LNP的稳定性,并规避严格的冷链要求.
研究的目的:
- 系统地研究各种辅助剂和缓冲剂对冷化mRNA-LNP的稳定性和性能的影响.
- 确定新的冷保护剂和缓冲系统,作为传统的基于糖的配方的替代品.
- 了解影响解热化mRNA-LNP功能的结构-活动关系.
主要方法:
- 选45种mRNA-LNP配方使用各种溶解保护剂 (寡糖,聚合物,氨基酸) 和缓冲剂.
- 使用高通量小角度X射线散射 (SAXS) 和低温传输电子显微镜 (cryo-TEM) 的物理化学和结构特征.
- 在人类外周血液单核细胞 (PBMCs) 中评估细胞转化疗效和先天免疫反应 (细胞因子分泌).
主要成果:
- 在Tris或酸盐缓冲器中确定了聚乙烯 (PVP) 作为有效的冷保护剂,在mRNA-LNP稳定性方面表现优于传统的寡糖类.
- 证明了mRNA-LNP结构特征和细胞翻译疗效后溶解后的复杂相互作用.
- 观察到细胞因子分泌特征的最小变化,表明冷化mRNA-LNP的免疫相容性得到保护.
结论:
- 解热化是一种稳定mRNA-LNP的可行策略,可以减少严格的存储和分配.
- 像PVP这样的新型辅助剂为开发稳定,功能性冷解mRNA-LNP配方提供了有希望的替代品.
- 这项研究为冷化mRNA-LNP疗法的合理设计提供了基础,通过将结构,稳定性和体外性能联系起来.
更多相关视频
08:10Author Spotlight: Enhancing Lipid Nanoparticle Formation Through Turbulent Mixing in Confined Geometries
Published on: August 23, 2024
3.5K
08:54Determining Four Components in a Lipid Nanoparticle RNA Delivery System by Liquid Chromatography Combined with Evaporative Light Scattering Detector
Published on: May 30, 2025
314
相关概念视频
Nuclear Export of mRNA
Before mRNAs are exported to the cytoplasm, it is crucial to check each mRNA for structural and functional integrity. Eukaryotic cells use several different mechanisms, collectively known as mRNA surveillance, to look for irregularities in mRNAs. Irregular or aberrant mRNA are rapidly degraded by various enzymes. If a defective mRNA escapes the surveillance, it would be translated into a protein which would either be non-functional or not function properly. One of the primary irregularities in...
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,...
Regulated mRNA Transport
In eukaryotes, transcription and translation are compartmentalized; an mRNA is first synthesized in the nucleus and then selectively transported to the cytoplasm for protein synthesis. Before transport, a pre-mRNA undergoes several steps of post-transcriptional modifications including splicing, 5' capping, and the addition of a poly-adenine tail. Various proteins bind to the pre-mRNA during these modifications. The mRNA transport takes place with the help of multiple proteins playing specific...
lncRNA - Long Non-coding RNAs
In humans, more than 80% of the genome gets transcribed. However, only around 2% of the genome codes for proteins. The remaining part produces non-coding RNAs which includes ribosomal RNAs, transfer RNAs, telomerase RNAs, and regulatory RNAs, among other types. A large number of regulatory non-coding RNAs have been classified into two groups depending upon their length – small non-coding RNAs, such as microRNA, which are less than 200 nucleotides in length, and long non-coding RNA (lncRNA)...
Nuclear Localization Signals and Import
Proteins targeted to the nucleus carry short stretches of amino acid sequences called the nuclear localization signal or NLS. Classical nuclear localization signals are of two types: monopartite and bipartite NLS. Monopartite classical NLS (cNLS) consists of a single cluster of 4-8 amino acids. Bipartite cNLS consists of two clusters of 2-3 amino acids and a 9-12 residue long proline-rich linker bridging the two clusters. Signal clusters are rich in positively charged amino acids such as...
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...
