Engineering Smart Protein Coronas to Advance Theraputic mRNA Delivery

虽然mRNA的基因治疗提供了巨大的潜力，以改变遗传管理， 然而，在疾病中，在网状内皮系统（例如肝脏和脾脏）外递送mRNA仍然具有挑战性。 如果没有能够到达肺、心脏、大脑或胰腺等器官的载体，mRNA的潜力将无法发挥。 治疗这些器官的疾病，如哮喘、冠状动脉疾病、多发性硬化症和糖尿病， 还未开发这项研究将建立蛋白质冠作为一种新的脂质纳米颗粒设计元素， 可以被工程化以实现网状内皮系统外的mRNA递送。在注射到 在体内，脂质纳米颗粒立即被局部蛋白质包裹，形成蛋白质冠， 细胞和组织。天然日冕的组成已经显示出强烈影响纳米颗粒的命运 in vivo.这项工作将通过用蛋白质预包被mRNA脂质纳米颗粒来开发“智能”蛋白质冠， 在给药前定位于网状内皮系统外的器官。 在目标1中，将评估天然存在的蛋白质的小文库指导mRNA表达的能力。 在C57 BL/6小鼠中，将其递送至非网状内皮器官，包括肺、心脏、脑和胰腺。 成像和流式细胞术将用于识别smart corona包被的mRNA脂质的器官和细胞靶点。 纳米粒子目的2将阐明智能冠状包被脂质纳米粒使 差异器官靶向。智能电晕可以通过改变循环时间， 募集额外的血清蛋白作为活性靶向剂，或通过结合特异性细胞表面 受体通过智能冠本身或通过额外吸附的蛋白质。蛋白质组分析将确定 蛋白质在静脉注射时吸附包被的脂质纳米颗粒，细胞受体阻断研究将 鉴定促进有效mRNA递送的细胞靶点。 这项工作是创新的，因为它将建立蛋白质冠作为一个以前不受重视的 纳米颗粒设计元件，其可以被工程化以使mRNA能够递送到具有挑战性的器官靶点。而 有效的智能冠状病毒将为研究人员提供新的工具，以推进药物靶向， 将为未来的智能电晕设计提供信息。这项研究意义重大，因为它将有助于范式转变 靶向策略，以加速肝细胞外疾病的mRNA治疗剂的临床转化。 此外，这项研究所获得的结果将促进核酸、小分子和 蛋白质从不同的药物载体，如纳米颗粒，药物缀合物和可植入装置。最后， 这项工作产生的知识有可能改变和加速有针对性的发展， 毒品携带者

While gene therapy with mRNA offers tremendous potential to transform the management of genetic diseases, delivering mRNA outside the reticuloendothelial system (e.g. liver and spleen) remains challenging. Without carriers that can reach organs such as the lungs, heart, brain, or pancreas, the potential of mRNA to treat diseases of those organs, such as asthma, coronary artery disease, multiple sclerosis, and diabetes, will remain untapped. This research will establish the protein corona as a new lipid nanoparticle design element that can be engineered to achieve mRNA delivery outside the reticuloendothelial system. Upon injection into the body, lipid nanoparticles are immediately coated in local proteins, forming a protein corona that interfaces with cells and tissues. The composition of that natural corona has been shown to strongly influence nanoparticle fate in vivo. This work will develop “smart” protein coronas by pre-coating mRNA lipid nanoparticles with proteins that localize to organs outside the reticuloendothelial system prior to administration. In Aim 1, a small library of naturally-occurring proteins will be evaluated for their ability to direct mRNA delivery to non-reticuloendothelial organs, including the lungs, heart, brain, and pancreas, in C57BL/6 mice. Imaging and flow cytometry will be used to identify organ and cellular targets of smart corona-coated mRNA lipid nanoparticles. Aim 2 will elucidate the mechanisms by which smart corona-coated lipid nanoparticles enable differential organ targeting. Smart coronas may enable differential targeting by altering circulation time, by recruiting additional serum proteins that serve as active targeting agents, or by binding specific cell surface receptors via the smart corona itself or via additionally adsorbed proteins. Proteomic analysis will determine what proteins adsorb coated-lipid nanoparticles upon intravenous injection, and cell receptor blocking studies will identify cellular targets that facilitate effective mRNA delivery. This work is innovative in that it will establish the protein corona as a previously-unappreciated nanoparticle design element that can be engineered to enable mRNA delivery to challenging organ targets. While efficacious smart coronas will provide researchers with new tools to advance drug targeting, mechanistic insights will inform future smart corona design. This research is significant because it will contribute a paradigm-shifting targeting strategy to accelerate the clinical translation of mRNA therapeutics for extrahepatocellular diseases. Further, the results obtained by this research will advance the delivery of nucleic acids, small molecules, and proteins from diverse drug carriers, such nanoparticles, drug conjugates, and implantable devices. Ultimately, the knowledge generated by this work has the potential to transform and accelerate the development of targeted drug carriers.