Scuderi Grazia, Arcidiacono Antonio, Cavalli Eugenio, Basile Maria Sofia, Nardo Antonella, Nicoletti Ferdinando, Fagone Paolo
Department of Biomedical and Biotechnological Sciences, University of Catania, 95123 Catania, Italy.
Department of Medicine and Surgery, "Kore" University of Enna, 94100 Enna, Italy.
Biomedicines. 2025 Aug 27;13(9):2090. doi: 10.3390/biomedicines13092090.
T-cell acute lymphoblastic leukemia (T-ALL) is an aggressive malignancy of immature T cells, driven by dysregulated transcriptional networks and oncogenic signaling pathways. Here, we present the first comprehensive analysis of the expression and regulation of TSPAN32, a tetraspanin implicated in lymphocyte homeostasis, in T-ALL. : Transcriptomic data from the Leukemia MILE study (GSE13159) were analyzed to assess TSPAN32 expression across leukemic subtypes. Gene Set Enrichment Analysis (GSEA) was performed to explore biological pathways associated with TSPAN32-correlated genes. For mechanistic validation, HPB-ALL cells were used as a model, with NOTCH signaling inhibited by γ-secretase inhibitor (GSI) treatment and TAL1-LMO1 overexpression induced through doxycycline-inducible lentiviral vectors. Gene expression changes were quantified by RT-qPCR. : TSPAN32 was frequently downregulated in T-ALL compared to healthy bone marrow, although expression was retained in a subset of cases. GSEA revealed that TSPAN32-correlated genes were inversely associated with cell cycle-related programs, consistent with its established role as a negative regulator of T cell proliferation. Mechanistically, TAL1-LMO1 overexpression strongly induced TSPAN32, while GSI-mediated NOTCH inhibition partially reactivated its expression. Interestingly, GSI treatment also increased TAL1 levels despite downregulating LMO1. Conversely, TAL1-LMO1 overexpression suppressed NOTCH1 and NOTCH3, highlighting a reciprocal regulatory interplay between NOTCH and TAL1/LMO1 oncogenic circuits that shapes TSPAN32 expression dynamics in T-ALL. : This study identifies TSPAN32 as a novel transcriptional target under the influence of key leukemogenic pathways and suggests its potential role as a modulator of leukemic T cell proliferation, with implications for therapeutic strategies targeting TAL1 and NOTCH signaling.
T 细胞急性淋巴细胞白血病（T-ALL）是一种由转录网络失调和致癌信号通路驱动的未成熟 T 细胞侵袭性恶性肿瘤。在此，我们首次对 TSPAN32 在 T-ALL 中的表达及调控进行了全面分析，TSPAN32 是一种与淋巴细胞稳态有关的四跨膜蛋白。：分析白血病 MILE 研究（GSE13159）的转录组数据，以评估 TSPAN32 在白血病亚型中的表达情况。进行基因集富集分析（GSEA）以探索与 TSPAN32 相关基因有关的生物学途径。为进行机制验证，使用 HPB-ALL 细胞作为模型，通过γ-分泌酶抑制剂（GSI）处理抑制 NOTCH 信号，并通过强力霉素诱导的慢病毒载体诱导 TAL1-LMO1 过表达。通过 RT-qPCR 定量基因表达变化。：与健康骨髓相比，TSPAN32 在 T-ALL 中经常下调，尽管在一部分病例中仍保留表达。GSEA 显示，TSPAN32 相关基因与细胞周期相关程序呈负相关，与其作为 T 细胞增殖负调节因子的既定作用一致。从机制上讲，TAL1-LMO1 过表达强烈诱导 TSPAN32，而 GSI 介导的 NOTCH 抑制部分重新激活其表达。有趣的是，尽管下调了 LMO1，但 GSI 处理也增加了 TAL1 水平。相反，TAL1-LMO1 过表达抑制了 NOTCH1 和 NOTCH3，突出了 NOTCH 与 TAL1/LMO1 致癌回路之间的相互调节相互作用塑造了 T-ALL 中 TSPAN32 的表达动态。：本研究确定 TSPAN32 是关键致白血病途径影响下的一个新转录靶点，并表明其作为白血病 T 细胞增殖调节剂的潜在作用，对靶向 TAL1 和 NOTCH 信号的治疗策略具有启示意义。
