Replacement of animal use in measuring cardiomyocyte response to drug safety profiling using a novel NMR metabolomics technology

用于治疗的药物的开发依赖于动物试验，以确保它们对人们使用是安全的。在过去，一些被认为是安全的药物会导致严重的心脏问题，甚至死亡。有害药物被发现会通过改变控制电活动和工作所需能量的分子的行为，最终阻止心脏正常跳动，从而阻止心脏正常运作。为此，所有开发的药物都必须经过对动物心脏有害影响的测试。我们的项目专注于一项相对较新的技术“核磁共振代谢组学”，它可以准确地识别心脏在细胞水平上使用能量的变化，使我们能够通过监测人类心脏细胞来研究药物的有害影响，而无需在小鼠（和其他动物）上进行实验。据估计，全球有35万只动物被用于药物测试。据我们计算，每年至少有1400只老鼠被用于发表的研究，专门用于测试对心脏的有害影响。然而，人类和动物的生物学差异意味着这些测试并不总是合适的。可能会错过对人类特定的潜在有害影响，或者检测到对人类无害的影响并丢弃药物。因此，在药物试验中使用的动物比需要的要多。为了解决这个问题，学术研究小组和制药公司已经开始使用人体细胞来检测和了解有害药物的作用。人类干细胞可以在实验室中转化为心脏细胞，已被证明是研究药物治疗后电活动和能量使用变化的热门选择。然而，缺乏足够灵敏的技术来检测导致心脏衰竭的细微而复杂的变化。因此，对用人体等效试验取代动物试验的信心一直是缓慢而有限的。我们的新型核磁共振代谢组学技术提供了比目前用于研究人类细胞能量使用的技术更先进的分析，能够以显著的灵敏度检测更多的信息。在这个项目中，我们将在人类干细胞制造的心脏细胞上测试核磁共振代谢组学技术。我们用有害的药物治疗心脏细胞，我们知道这些药物会导致电活动和能量使用的变化。我们还操纵心脏细胞，使它们对药物的作用更敏感。这模仿了现实生活中的情况，有些人在对有害药物作出反应后有更大的心力衰竭风险。我们已经从因心脏病而受损的人类患者干细胞中提取了数据，并利用这些数据验证了结果。我们估计，利用核磁共振代谢组学技术和干细胞可以取代多达50%的动物，这些动物将被用于测试药物对心脏的有害影响。此外，加深对药物如何造成有害影响的了解有助于改进使用动物的其他试验的设计，从而进一步减少。为了实现我们的3Rs影响，我们在项目过程中与被确定为该技术最终用户的学术研究小组和制药公司合作，以充分了解他们进行的动物试验以及他们如何使用该核磁共振代谢组学技术。我们还创建数字培训材料和研讨会活动，以支持在干细胞中作为动物替代技术的NMR代谢组学的吸收和使用。在这个项目之外，我们预计干细胞核磁共振代谢组学的潜力将推动未来的应用，取代其他疾病和测试环境中使用的动物试验。

The development of drugs for therapeutic treatment depends on animal testing to ensure that they are safe for people to use. In the past some drugs that were thought to be safe went on to cause serious heart problems, and even fatalities. Harmful drugs were found to stop the heart from functioning properly by altering the behaviour of molecules that control the electrical activity and energy required to work, and ultimately the ability of the heart to beat properly. In response to this, all drugs that are developed must be tested for harmful effects on animal hearts. Our project is focused on a relatively new technology 'NMR metabolomics', which can accurately identify changes in how the heart uses energy at the cell level, allowing us to study the harmful effects of drugs by monitoring human heart cells without the need to perform experiments on mice (and other animals).It is estimated that 350,000 animals are used globally in drug testing. We have calculated that at least 1,400 mice are used in published studies each year, specifically for testing harmful effects on the heart. However, differences in the biology of humans and animals means that these tests are not always appropriate. Potentially harmful effects specific to humans might be missed, or else effects not harmful in humans are detected and the drug discarded. Therefore, more animals are used in drug tests than are needed.To address this, academic research groups and Pharma have begun to use human cells to detect and understand harmful drug effects. Human stem cells, which can be turned into heart cells in the laboratory, have proved a popular choice for studying changes in electrical activity and energy use in response to drug treatment. However, there is a lack of technologies that are sensitive enough to detect the subtle and complex changes that cause the heart to fail. Confidence in the replacement of animal tests with human equivalents has therefore been slow and limited. Our novel implementation of NMR metabolomics technology offers a more advanced analysis than the current techniques being used to study energy use in human cells, being capable of detecting much more information with significant sensitivity. In this project we will test NMR metabolomics technology on heart cells that we make from human stem cells. We treat the heart cells with harmful drugs that we know cause changes in electrical activity and energy use. We also manipulate the heart cells so that they are more sensitive to the effects of the drugs. This mimics the real-life situation where some people have a greater risk of heart failure in response to harmful drugs. The results are validated using data that we have already produced from human patient stem cells that are damaged due to heart disease.We estimate that employing NMR metabolomics technology with stem cells could replace up to 50% of the animals that would be used to test drugs for harmful effects on the heart. In addition, increased understanding of how drugs cause harmful effects helps to improve design for other tests in which animals are used, leading to further reduction. To achieve our 3Rs impact, we are during the course of the project engaging with academic research groups and Pharma who are identified as being end users of the technology in order to understand fully the animal tests that they perform and how they would use this NMR metabolomics technology. We also create digital training materials and workshop activities that support uptake and use of NMR metabolomics in stem cells as an animal replacement technology. Beyond this project we anticipate that demonstration of the potential for NMR metabolomics with stem cells will drive future application as a replacement for animal tests used in other disease and testing settings.