Krüger Dustin N, Bosman Matthias, Van Assche Charles X L, Wesley Callan D, Cillero-Pastor Berta, Delrue Leen, Heggermont Ward, Bartunek Jozef, De Meyer Guido R Y, Van Craenenbroeck Emeline M, Guns Pieter-Jan, Franssen Constantijn
Laboratory of Psychopharmacology, Faculty of Medicine and Health Sciences, Faculty of Pharmaceutical, Biomedical and Veterinary Sciences, Campus Drie Eiken, University of Antwerp, Universiteitsplein 1, Antwerp, B-2610, Belgium.
Division M4I - Imaging Mass Spectrometry (IMS), Faculty of Health, Medicine and Life Sciences, Maastricht MultiModal Molecular Imaging Institute, Maastricht University, Universiteitssingel 50, Maastricht, 6229 ER, The Netherlands.
Cardiooncology. 2024 Jun 22;10(1):40. doi: 10.1186/s40959-024-00241-1.
The anthracycline doxorubicin (DOX) is a highly effective anticancer agent, especially in breast cancer and lymphoma. However, DOX can cause cancer therapy-related cardiovascular toxicity (CTR-CVT) in patients during treatment and in survivors. Current diagnostic criteria for CTR-CVT focus mainly on left ventricular systolic dysfunction, but a certain level of damage is required before it can be detected. As diastolic dysfunction often precedes systolic dysfunction, the current study aimed to identify functional and molecular markers of DOX-induced CTR-CVT with a focus on diastolic dysfunction.
Male C57BL/6J mice were treated with saline or DOX (4 mg/kg, weekly i.p. injection) for 2 and 6 weeks (respectively cumulative dose of 8 and 24 mg/kg) (n = 8 per group at each time point). Cardiovascular function was longitudinally investigated using echocardiography and invasive left ventricular pressure measurements. Subsequently, at both timepoints, myocardial tissue was obtained for proteomics (liquid-chromatography with mass-spectrometry). A cohort of patients with CTR-CVT was used to complement the pre-clinical findings.
DOX-induced a reduction in left ventricular ejection fraction from 72 ± 2% to 55 ± 1% after 2 weeks (cumulative 8 mg/kg DOX). Diastolic dysfunction was demonstrated as prolonged relaxation (increased tau) and heart failure was evident from pulmonary edema after 6 weeks (cumulative 24 mg/kg DOX). Myocardial proteomic analysis revealed an increased expression of 12 proteins at week 6, with notable upregulation of SERPINA3N in the DOX-treated animals. The human ortholog SERPINA3 has previously been suggested as a marker in CTR-CVT. Upregulation of SERPINA3N was confirmed by western blot, immunohistochemistry, and qPCR in murine hearts. Thereby, SERPINA3N was most abundant in the endothelial cells. In patients, circulating SERPINA3 was increased in plasma of CTR-CVT patients but not in cardiac biopsies.
We showed that mice develop heart failure with impaired systolic and diastolic function as result of DOX treatment. Additionally, we could identify increased SERPINA3 levels in the mice as well as patients with DOX-induced CVT and demonstrated expression of SERPINA3 in the heart itself, suggesting that SERPINA3 could serve as a novel biomarker.
蒽环类药物阿霉素（DOX）是一种高效的抗癌药物，尤其在乳腺癌和淋巴瘤治疗中应用广泛。然而，DOX在治疗期间及治疗后的幸存者中可导致癌症治疗相关的心血管毒性（CTR-CVT）。目前CTR-CVT的诊断标准主要侧重于左心室收缩功能障碍，但在其能够被检测到之前需要一定程度的损伤。由于舒张功能障碍通常先于收缩功能障碍出现，本研究旨在确定DOX诱导的CTR-CVT的功能和分子标志物，重点关注舒张功能障碍。
雄性C57BL/6J小鼠分别用生理盐水或DOX（4mg/kg，每周腹腔注射）处理2周和6周（累积剂量分别为8mg/kg和24mg/kg）（每个时间点每组n = 8）。使用超声心动图和有创左心室压力测量纵向研究心血管功能。随后，在两个时间点获取心肌组织进行蛋白质组学分析（液相色谱-质谱联用）。使用一组CTR-CVT患者补充临床前研究结果。
DOX处理2周后（累积8mg/kg DOX），左心室射血分数从72±2%降至55±1%。舒张功能障碍表现为舒张期延长（tau增加），6周后（累积24mg/kg DOX）出现肺水肿，心力衰竭明显。心肌蛋白质组学分析显示第6周有12种蛋白质表达增加，DOX处理组动物中SERPINA3N显著上调。人类同源蛋白SERPINA3此前已被认为是CTR-CVT的一个标志物。通过蛋白质免疫印迹、免疫组织化学和qPCR在小鼠心脏中证实了SERPINA3N的上调。因此，SERPINA3N在内皮细胞中含量最丰富。在患者中，CTR-CVT患者血浆中循环SERPINA3增加，但在心脏活检组织中未增加。
我们发现DOX处理导致小鼠出现收缩和舒张功能受损的心力衰竭。此外，我们在小鼠以及DOX诱导的CVT患者中发现SERPINA3水平升高，并证明SERPINA3在心脏本身表达，这表明SERPINA3可能是一种新型生物标志物。
