Kverneland Anders Handrup, Østergaard Ole, Sohlin Joel Emanuel, Noringriis Inge Mansfield, Jurlander Rebecca S, Olsen Jesper Velgaard, Svane Inge Marie
Department of Oncology, National Center of Cancer Immune Therapy (CCIT-DK), Copenhagen University Hospital, Herlev, Hovedstaden, Denmark.
NNF Center for Protein Research, University of Copenhagen, Kobenhavn, Denmark.
BMJ Oncol. 2025 Jun 6;4(1):e000696. doi: 10.1136/bmjonc-2024-000696. eCollection 2025.
Immune-related adverse events (irAEs) are a growing challenge with checkpoint inhibitors (CPIs) and are complicated by the lack of suitable response biomarkers in many irAEs. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) based proteomics is a system-wide unbiased analysis method suited for biomarker discovery. In this study, plasma samples from patients suffering from irAEs were analysed using MS-based proteomics.
The discovery cohort consisted of 37 melanoma patients experiencing an irAE (colitis, hepatitis and nephritis) collected around irAE management. The validation cohort consisted of 34 irAE patients (colitis, hepatitis, nephritis and pneumonitis) at management, with a control group consisting of 34 patients treated with CPIs without an irAE. The plasma samples were processed with bottom-up proteomics and analysed on an Orbitrap Astral Mass Spectrometer with short LC gradients.
During an irAE, we found upregulation of multiple acute-phase protein reactants. The level of these was higher in patients in combination CPI therapy-also observed in the control cohort without irAE. We found multiple hepatic proteins associated with immune-related hepatitis, including fructose-biphosphate aldolase B (ALDOB), showing the widest dynamic range. Further, we found a correlation of lipocalin-2/neutrophil gelatinase-associated lipocalin (LCN2/NGAL) to nephritis and colitis with a significant correlation to the clinical toxicity grade confirmed by an immunoassay. Finally, high levels of angiotensinogen (AGT), chitinase-3-like protein 1 (CHI3L1) and lectin mannose-binding 2 (LMAN2) showed a significant association with poor prednisolone response.
In conclusion, using LC-MS/MS-based plasma proteomics, we identified several irAE-related biomarker candidates to be further assessed for toxicity grading and management in the context of monitoring irAEs.
免疫相关不良事件（irAE）是检查点抑制剂（CPI）面临的一个日益严峻的挑战，且在许多irAE中因缺乏合适的反应生物标志物而变得复杂。基于液相色谱 - 串联质谱（LC - MS/MS）的蛋白质组学是一种适用于生物标志物发现的全系统无偏分析方法。在本研究中，我们使用基于质谱的蛋白质组学分析了患有irAE患者的血浆样本。
发现队列由37名在irAE管理期间出现irAE（结肠炎、肝炎和肾炎）的黑色素瘤患者组成。验证队列由34名在管理时出现irAE（结肠炎、肝炎、肾炎和肺炎）的患者组成，对照组由34名接受CPI治疗但未出现irAE的患者组成。血浆样本采用自下而上的蛋白质组学方法进行处理，并在具有短液相色谱梯度的Orbitrap Astral质谱仪上进行分析。
在irAE期间，我们发现多种急性期蛋白反应物上调。这些蛋白在联合使用CPI治疗的患者中水平更高，在无irAE的对照队列中也观察到了这一现象。我们发现了多种与免疫相关肝炎相关的肝脏蛋白，包括果糖 - 双磷酸醛缩酶B（ALDOB），其动态范围最广。此外，我们发现脂质运载蛋白2/中性粒细胞明胶酶相关脂质运载蛋白（LCN2/NGAL）与肾炎和结肠炎相关，并且通过免疫测定证实其与临床毒性分级有显著相关性。最后，高水平的血管紧张素原（AGT）、几丁质酶3样蛋白1（CHI3L1）和凝集素甘露糖结合2（LMAN2）与泼尼松龙反应不佳显著相关。
总之，通过基于LC - MS/MS的血浆蛋白质组学，我们鉴定了几种与irAE相关的生物标志物候选物，有待在监测irAE的背景下进一步评估其用于毒性分级和管理的价值。
