Data_Sheet_1_RELSA—A multidimensional procedure for the comparative assessment of well-being and the quantitative determination of severity in experimental procedures.pdf

Good science in translational research requires good animal welfare according to the principles of 3Rs. In many countries, determining animal welfare is a mandatory legal requirement, implying a categorization of animal suffering, traditionally dominated by subjective scorings. However, how such methods can be objectified and refined to compare impairments between animals, subgroups, and animal models remained unclear. Therefore, we developed the RELative Severity Assessment (RELSA) procedure to establish an evidence-based method based on quantitative outcome measures such as body weight, burrowing behavior, heart rate, heart rate variability, temperature, and activity to obtain a relative metric for severity comparisons. The RELSA procedure provided the necessary framework to get severity gradings in TM-implanted mice, yielding four distinct RELSA thresholds L1<0.27, L2<0.59, L3<0.79, and L4<3.45. We show further that severity patterns in the contributing variables are time and model-specific and use this information to obtain contextualized between animal-model and subgroup comparisons with the severity of sepsis > surgery > restraint stress > colitis. The bootstrapped 95% confidence intervals reliably show that RELSA estimates are conditionally invariant against missing information but precise in ranking the quantitative severity information to the moderate context of the transmitter-implantation model. In conclusion, we propose the RELSA as a validated tool for an objective, computational approach to comparative and quantitative severity assessment and grading. The RELSA procedure will fundamentally improve animal welfare, data quality, and reproducibility. It is also the first step toward translational risk assessment in biomedical research.

转化研究中的优质科学实践需遵循3R（3Rs）原则以保障良好的动物福利。在诸多国家，动物福利评估属于法定强制要求，这意味着需对动物痛苦程度进行分级，而传统分级方法多以主观评分法为主。然而，如何将此类方法客观化、精细化，以实现不同动物、亚组及动物模型间的损伤程度比较，此前尚未明确。为此，本研究开发了相对严重程度评估（RELative Severity Assessment, RELSA）流程，基于体重、掘洞行为、心率、心率变异性、体温及活动度等定量结局指标，构建循证方法以获取用于严重程度比较的相对量化指标。RELSA流程为经发射器（TM）植入的小鼠提供了严重程度分级的必要框架，最终得到四个明确的RELSA阈值：L1<0.27、L2<0.59、L3<0.79及L4<3.45。研究进一步发现，各贡献变量的严重程度模式具有时间特异性与模型特异性，并基于此实现了不同动物模型及亚组间的情境化比较，其严重程度排序为：脓毒症>手术>束缚应激>结肠炎。通过Bootstrap自助法得到的95%置信区间可靠证实：RELSA评估结果在存在信息缺失时仍具备条件不变性，且在发射器植入模型的中度情境下，其对定量严重程度信息的排序精度良好。综上，本研究提出RELSA可作为经验证的工具，用于实现客观、可计算的比较式定量严重程度评估与分级。RELSA流程将从根本上提升动物福利、数据质量与研究可重复性，同时也是生物医学研究中转化风险评估的首个可行步骤。