Background/Objectives: Hodgkin’s lymphoma (HL) affects 2–4 individuals per 100,000 annually. Standard treatment includes radiotherapy and ABVD chemotherapy, achieving a 95% survival rate. However, HL survivors face an elevated risk of treatment-related morbidity, particularly the development of secondary malignancies. Previous studies have demonstrated that ABVD treatment induces a high frequency of chromosomal aberrations (CAs) in lymphocytes from HL patients, with higher frequencies one year after treatment than during treatment. This study aimed to determine whether HL treatment also induces unclassified chromosomal/nuclear aberrations (UnCAs) in the lymphocytes of HL patients, and whether these alterations may serve as complementary indicators of genomic instability.Methods: Peripheral blood lymphocytes from HL patients were collected at three time points: before treatment (BT), during treatment (DT), and one year after treatment (1yAT) with ABVD chemotherapy and radiotherapy. A minimum of 3000 nuclei were analyzed per patient to identify and quantify UnCAs. These results were compared to UnCA frequencies in healthy individuals.Results: The percentage of cells presenting UnCAs per 3000 nuclei was 23.92% BT, 18.58% DT, and 30.62% 1yAT. All values were significantly higher (p< 0.016) than the 8.16% observed in healthy controls. The increase was primarily driven by free chromatin and micronuclei clusters. UnCA frequency was lower during treatment than one year after, likely due to the elimination of highly damaged cells through apoptosis or lack of proliferative capacity. Over time, however, persistent genomic damage appears to accumulate in surviving cells, becoming more evident post-treatment. A parallel trend was observed between the frequencies of UnCAs free chromatin, micronucleus and micronuclei clusters, and classical CAs, showing a similar pattern of genomic damage induced by therapy.Conclusions: The post-treatment increase in UnCAs indicates ongoing genomic instability, possibly driven by the selective survival of hematopoietic stem cells with higher genomic fitness. Given their persistence and association with therapy-induced damage, free chromatin and micronuclei clusters may serve as early biomarkers for secondary cancer risk in HL survivors.
背景/目的：霍奇金淋巴瘤（HL）的年发病率为每10万人中2-4例。标准治疗方案包括放疗和ABVD化疗，可使患者生存率达到95%。然而，HL幸存者面临治疗相关并发症风险升高的问题，尤其是继发性恶性肿瘤的发生。既往研究表明，ABVD治疗会诱导HL患者淋巴细胞出现高频染色体畸变（CAs），且治疗一年后的畸变频率高于治疗期间。本研究旨在探讨HL治疗是否同样会诱导患者淋巴细胞出现未分类染色体/核畸变（UnCAs），以及这些改变是否可作为基因组不稳定性的补充指标。 方法：在ABVD化疗联合放疗的三个时间点采集HL患者外周血淋巴细胞：治疗前（BT）、治疗期间（DT）及治疗后一年（1yAT）。每位患者至少分析3000个细胞核以识别和量化UnCAs，并将结果与健康个体的UnCA频率进行比较。 结果：每3000个细胞核中呈现UnCAs的细胞百分比分别为：治疗前23.92%、治疗期间18.58%、治疗后一年30.62%。所有数值均显著高于健康对照组的8.16%（p<0.016）。该增长主要由游离染色质和微核簇驱动。治疗期间的UnCA频率低于治疗后一年，这可能是由于高度损伤细胞通过凋亡或增殖能力缺失被清除。然而随着时间的推移，持续存在的基因组损伤似乎在存活细胞中累积，并在治疗后表现得更为明显。游离染色质、微核及微核簇等UnCAs的频率与经典CAs呈现平行变化趋势，显示出治疗诱导基因组损伤的相似模式。 结论：治疗后UnCAs的增加表明基因组不稳定性持续存在，这可能是具有较高基因组适应性的造血干细胞选择性存活的结果。鉴于游离染色质和微核簇的持续性及其与治疗损伤的关联性，它们或可作为HL幸存者继发性癌症风险的早期生物标志物。
肿瘤(癌症)患者之家