mRNA-LNPs for ARDS

摘要/项目总结 尽管对急性呼吸窘迫综合征（ARDS）的途径进行了50多年的研究， 仍然没有药物可以改善其死亡率。从药理学的角度来看，这种缺乏临床试验成功 福尔斯分为两个主要方面：药物输送到肺泡的能力差，没有平台技术可以轻松设计药物 对于给定的目标蛋白质。在这里，我们的目标是用一种纳米技术解决这些问题。我们首先 开发纳米级药物载体（nanocarrier），可以在肺泡中大量浓缩药物， 约300倍，IV注射后。这些纳米载体是脂质纳米颗粒（LNP），其与靶向药物缀合。 将LNP导向肺泡内皮细胞（通过LNP上的抗PECAM抗体 表面）或肺泡边缘白细胞（通过我们最近开发的NAP标签）。虽然我们多年来 使用这些纳米载体来递送小分子药物，这类货物药物几乎没有分子， 影响了ARDS相关通路，药物难以加载到LNP中。因此，在这里，我们将为 第一次在我们的肺泡靶向LNP内提供一类新的药物， 途径：修饰的mRNA。修饰的mRNA-LNP驱动编码蛋白的表达约48小时， 次给药结束在这项提案中，我们将联合收割机结合我们的肺泡靶向和mRNA技术来治疗两个最大的 早期-中期ARDS的病理过程：肺泡毛细血管渗漏（Aim 1，靶向内皮细胞）和 白细胞浸润（目标2，靶向肺泡边缘白细胞）。对于这2个与ARDS相关的 疾病过程中，我们将递送编码分泌分子（Ang 1或IL-10）或 细胞内分子（VE-钙粘蛋白或IkB），以比较这些不同的蛋白质类如何与此 技术.最后，我们将在两个ARDS样小鼠模型中测试这些（目的1和2），并在体外进行实验。 人肺（目标3）。这里开发的平台技术可以直接生产ARDS治疗剂， 并且以后还可以扩展到探测ARDS病理生理学和治疗其它肺泡疾病。

ABSTRACT / PROJECT SUMMARY Despite 50+ years of dissecting the pathways of acute respiratory distress syndrome (ARDS), there are still no drugs which improve its mortality. From a pharmacology perspective, this lack of clinical trial success falls into 2 major buckets: poor drug delivery to the alveoli and no platform technology to easily design a drug for a given target protein. Here, we aim to solve these problems with a single nanotechnology. We began by developing nano-scale drug carriers (nanocarriers) that can massively concentrate drugs in the alveoli, ~300-fold, after IV injection. These nanocarriers are lipid nanoparticles (LNPs) that are conjugated to targeting moieties that either direct the LNPs to alveolar endothelial cells (via an anti-PECAM antibody on the LNP surface), or to alveolar marginated leukocytes (via our recently developed NAP-tag). While we have for years used these nanocarriers to deliver small molecule drugs, that class of cargo drugs had few molecules that impacted ARDS-related pathways, and the drugs were difficult to load into LNPs. Therefore, here we will for the first time deliver inside our alveolar-targeted LNPs a new class of drugs that can target virtually any pathway: modified mRNA. Modified mRNA-LNPs drive the expression of encoded proteins for ~48 hours per dose. In this proposal, we will combine our alveolar-targeting & mRNA technologies to treat two of the biggest pathological processes of early-mid ARDS: alveolar capillary leak (Aim 1, targeting endothelial cells) and leukocyte infiltration (Aim 2, targeting alveolar marginated leukocytes). For each of these 2 ARDS-related disease processes, we will deliver mRNAs that encode either a secreted molecule (Ang1 or IL-10) or an intracellular molecule (VE-cadherin or IkB), to compare how these different protein classes function with this technology. Finally, we will test these in two ARDS-like mouse models of ARDS (Aims 1 & 2), and in ex vivo human lungs (Aim 3). The platform technology developed here may directly produce an ARDS therapeutic, and may also be extended later to probe ARDS pathophysiology, and treat other alveolar diseases.