Department of Pharmacology, The Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA; Department of Bioengineering, School of Engineering and Applied Sciences, University of Pennsylvania, Philadelphia, PA, USA.
Department of Pharmacology, The Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA.
Int J Pharm. 2023 Oct 15;645:123369. doi: 10.1016/j.ijpharm.2023.123369. Epub 2023 Sep 9.
Infusion reactions are a major risk for advanced therapeutics (e.g., engineered proteins nanoparticles, etc.), which can trigger the complement cascade, anaphylaxis, and other life-threatening immune responses. However, during the early phases of development, it is uncommon to assess for infusion reactions, given the labor involved in measuring multiple physiological parameters in rodents. Therefore, we sought to develop an automated quantification of rodent locomotion to serve as a sensitive screening tool for infusion reactions, with minimal added labor-time for each experiment. Here we present the detailed methods for building a motion tracking cage for mice, requiring ∼$100 of materials, ∼2 h to build and set up completely, and employing freely available software (DeepLabCut). The distance-walked after injection was first shown to have the predicted effects for stimulants (caffeine), sedatives (ketamine), and toxins (lipopolysaccharide). Additionally, the distance-walked more sensitively detected the effects of these compounds than did pulse oximetry-based measurements of the classical vital signs of heart rate, respiratory rate, and blood oxygen saturation. Finally, we examined a nanomedicine formulation that has been in preclinical development, liposomes targeted to the cell adhesion molecule ICAM. While this formulation has been studied across dozens of publications, it has not previously been noted to produce an infusion reaction. However, the automated motion tracking cage showed that ICAM-liposomes markedly reduce the distance-walked, which we confirmed by measuring the other vital signs. Importantly, the motion tracking cage added < 5 min of labor time per 5-mouse condition, while pulse oximetry with a neck cuff (by far the most stable oximetry signal in mice) required ∼ 100 min of labor time. Thus, automated measurement of distance-walked can indeed serve as a "sixth vital sign" for detecting infusion reactions during preclinical testing. Additionally, the device to measure distance-walked is easy and cheap to build and requires negligible labor time for each experiment, enabling distance-walked to be recorded in nearly every infusion experiment.
输注反应是高级治疗方法（例如工程蛋白纳米颗粒等）的主要风险，可能会引发补体级联反应、过敏反应和其他危及生命的免疫反应。然而，在开发的早期阶段，由于在啮齿动物中测量多个生理参数涉及到大量的工作，因此通常不评估输注反应。因此，我们试图开发一种自动量化啮齿动物运动的方法，作为一种敏感的输注反应筛选工具，每个实验的附加工作量最小。在这里，我们介绍了构建用于小鼠的运动跟踪笼的详细方法，该方法需要大约 100 美元的材料，大约 2 小时即可完全构建和设置完成，并使用免费提供的软件（DeepLabCut）。注射后的行走距离首先显示出对兴奋剂（咖啡因）、镇静剂（氯胺酮）和毒素（脂多糖）的预期影响。此外，与基于脉搏血氧仪的心率、呼吸率和血氧饱和度等经典生命体征的测量相比，行走距离更灵敏地检测到这些化合物的作用。最后，我们研究了一种处于临床前开发阶段的纳米医学制剂，即靶向细胞粘附分子 ICAM 的脂质体。虽然这种制剂已经在数十篇出版物中进行了研究，但以前没有注意到它会产生输注反应。然而，自动运动跟踪笼显示，ICAM-脂质体显著减少了行走距离，我们通过测量其他生命体征证实了这一点。重要的是，与颈带脉搏血氧仪（迄今为止在小鼠中最稳定的血氧仪信号）相比，运动跟踪笼每 5 只小鼠条件仅增加了<5 分钟的工作量，而脉搏血氧仪需要大约 100 分钟的工作量。因此，自动测量行走距离确实可以作为临床前测试中检测输注反应的“第六生命体征”。此外，用于测量行走距离的设备易于构建且每个实验所需的工作量可忽略不计，从而可以在几乎每个输注实验中记录行走距离。
