Engineering Smart Protein Coronas to Advance Theraputic mRNA Delivery

同时，基因治疗提供了巨大的潜力，以改变基因的管理 对于疾病，在网状内皮系统(如肝和脾)之外传递信使核糖核酸仍然具有挑战性。 如果没有可以到达肺、心脏、脑或胰腺等器官的载体，mRNA就有可能 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，将 保持未开发状态。这项研究将建立蛋白质冠层作为一种新的脂类纳米颗粒设计元件 可以被改造成在网状内皮系统之外实现信使核糖核酸的传递。在注入到 在体内，脂质纳米粒立即被包裹在局部蛋白质上，形成蛋白质冠层，与 细胞和组织。天然电晕的组成已被证明对纳米粒子的命运有很大影响 在活体内。这项工作将通过在信使核糖核酸脂纳米粒上预包覆具有以下特性的蛋白质来开发“智能”蛋白质冠冕 在给药前定位于网状内皮系统以外的器官。 在目标1中，将对自然产生的蛋白质的一个小文库进行评估，以确定它们引导mrna的能力。 C57BL/6小鼠的非网状内皮器官，包括肺、心、脑和胰腺。 成像和流式细胞术将用于鉴定SMART电晕包被的mRNA脂质的器官和细胞靶点 纳米粒子。目标2将阐明智能电晕涂层脂质纳米粒使 不同的器官靶向。智能日冕可以通过改变循环时间、通过 招募更多的血清蛋白作为活性靶向试剂，或通过结合特定的细胞表面 通过智能电晕本身或通过额外吸附的蛋白质的受体。蛋白质组分析将决定 蛋白质吸附包被脂质纳米粒静脉注射，细胞受体阻断研究将 确定促进有效信使核糖核酸传递的细胞靶点。 这项工作的创新之处在于它将把蛋白质日冕建立为一种以前未被认识到的 纳米颗粒设计元素，可以被设计成能够将信使RNA传递到具有挑战性的器官靶标。而当 有效的智能日冕将为研究人员提供新的工具来推进药物靶向、机械洞察力 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的转变 以靶向策略加速肝外疾病信使核糖核酸治疗药物的临床翻译。 此外，这项研究取得的成果将推动核酸、小分子和 来自不同药物载体的蛋白质，如纳米颗粒、药物结合物和可植入设备。最终， 这项工作所产生的知识有可能转化和加速有针对性的 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.