Wei Renrong, Qiu Dan, Zeng Xiangxing
Medical laboratory, Heyuan Key Laboratory of Molecular Diagnosis & Disease Prevention and Treatment, Doctors Station of Guangdong province, Heyuan People's Hospital, No.733 Wenxiang Road, Yuancheng District, Heyuan, Guangdong, China.
Department of Traditional Chinese Medicine, Heyuan People's Hospital, Heyuan, Guangdong, China.
Ann Hematol. 2025 Sep 16. doi: 10.1007/s00277-025-06605-6.
Approximately 1.5% of individuals with hemoglobin disorders carry the β-thalassemia gene variant, impacting around 40,000 newborns annually. Given the incomplete understanding of β-thalassemia pathogenesis, there is an urgent need to identify effective biomarkers to advance research, diagnosis, and treatment. This study aims to identify potential biomarkers for two key purposes: (1) diagnosing transfusion-dependent β-thalassemia (TDT) and (2) detecting iron overload complications, with a focus on functional markers that reflect iron metabolism dysregulation in TDT. This study integrates transcriptomic data from the Genome Sequence Archive dataset (CRA003639) with bioinformatics analysis to identify potential biomarkers associated with β-thalassemia. Subsequently, Hbb-bs and Hbb-bt double knockout mice were used to establish a β-thalassemia model, while C57BL/6JCya mice served as the control group, to validate the identified biomarkers through animal experiments. Seventeen reliable cell subsets were identified through rigorous annotation and screening. Quantitative analysis revealed a decreased proportion of immune cells (natural killer [NK] cells, T cells, macrophages, neutrophils, and monocytes) and an increased proportion of erythroid cells in the β-thalassemia group. Cell subset analysis focused on subsets that closely communicated with erythroid cells. Enrichment analysis of driver genes in these subsets revealed iron metabolism-related pathways in Erythroid_02 and Erythroid_03, and a ferroptosis-related pathway in Erythroid_05. Thalassemia model mice exhibited stronger iron ion fluorescence signals in primary hepatocytes, increased levels of total iron, Fe, and Fe in liver tissue, and decreased serum iron (SI) levels, indicating iron metabolism disorders. Reverse transcription polymerase chain reaction (RT-PCR) results showed differential gene expression, with BCL2L1, Hepb1, and Prdx6 downregulated and Spta1 and Snca upregulated in the TDT model group. This study comprehensively characterizes TDT at the cellular and molecular levels, offering insights into its pathogenesis and identifying potential therapeutic targets.
约1.5%的血红蛋白疾病患者携带β地中海贫血基因变异，每年影响约40000名新生儿。鉴于对β地中海贫血发病机制的了解尚不完整，迫切需要识别有效的生物标志物以推动研究、诊断和治疗。本研究旨在识别潜在生物标志物，以实现两个关键目标：（1）诊断输血依赖型β地中海贫血（TDT），（2）检测铁过载并发症，重点关注反映TDT中铁代谢失调的功能标志物。本研究将来自基因组序列存档数据集（CRA003639）的转录组数据与生物信息学分析相结合，以识别与β地中海贫血相关的潜在生物标志物。随后，使用Hbb-bs和Hbb-bt双敲除小鼠建立β地中海贫血模型，而C57BL/6JCya小鼠作为对照组，通过动物实验验证所识别的生物标志物。通过严格的注释和筛选，识别出17个可靠的细胞亚群。定量分析显示，β地中海贫血组中免疫细胞（自然杀伤细胞、T细胞、巨噬细胞、中性粒细胞和单核细胞）比例降低，红系细胞比例升高。细胞亚群分析聚焦于与红系细胞密切通讯的亚群。对这些亚群中驱动基因的富集分析揭示了红系_02和红系_03中铁代谢相关途径，以及红系_05中与铁死亡相关的途径。地中海贫血模型小鼠原代肝细胞中铁离子荧光信号更强，肝组织中总铁、铁和铁水平升高，血清铁水平降低，表明存在铁代谢紊乱。逆转录聚合酶链反应（RT-PCR）结果显示基因表达存在差异，TDT模型组中BCL2L1、Hepb1和Prdx6下调，Spta1和Snca上调。本研究在细胞和分子水平上全面表征了TDT，为其发病机制提供了见解，并识别出潜在的治疗靶点。
