Lainscsek Xenia, Kong Weibo, Rütgen Barbara C, Beck Julia, Brenig Bertram, Nolte Ingo, Murua Escobar Hugo, Taher Leila
Institute of Biomedical Informatics, Graz University of Technology, Graz, Austria.
Clinic for Hematology, Oncology and Palliative Care, Rostock University Medical Center, University of Rostock, Rostock, Germany.
Front Vet Sci. 2025 Apr 25;12:1577028. doi: 10.3389/fvets.2025.1577028. eCollection 2025.
B-cell receptor (BCR) signaling has revealed itself as a critical pathway in the pathogenesis of B-cell lymphoma. Within this pathway, the inhibition of Bruton's tyrosine kinase (BTK) or Phosphoinositide 3-kinases (PI3Ks) alone presents encouraging efficacy in the treatment of certain both canine and human hematological malignancies.
Here we characterized the effects of the BTK inhibitor Ibrutinib and the PI3K inhibitor AS-605240 as single and combined agents in the canine pre-clinical diffuse large B cell lymphoma (DLBCL) model CLBL-1 by assaying cell proliferation and metabolic activity, and performing RNA-seq to measure gene expression changes.
We found 2,336 differentially expressed genes (DEGs) across all treatment types and time points relative to the control. The largest number of DEGs were induced by the combination of Ibrutinib and AS-605240. These genes were involved in adaptive immune response, leukotriene D4 metabolic and terms related to regulation of GTP and GTPase mediated signal transduction. Weighted gene co-expression network analysis (WGCNA) detected nine gene modules, five of which were associated with treatment response. Eighteen-percent of genes within these modules were also differentially expressed. Notably, we observed one module that was exclusively associated with the combined treatment whose gene members were related to cellular metabolism, homeostasis signaling, and protein synthesis and regulation.
Narrowing in on highly connected genes of modules associated with treatment response with large fold changes across treatments which play roles in the main targeted pathways identified , and as potential primary candidates of the synergistic treatment effect.
B细胞受体（BCR）信号通路已成为B细胞淋巴瘤发病机制中的关键途径。在该通路中，单独抑制布鲁顿酪氨酸激酶（BTK）或磷酸肌醇3激酶（PI3K）在治疗某些犬类和人类血液系统恶性肿瘤方面显示出令人鼓舞的疗效。
在此，我们通过检测细胞增殖和代谢活性，并进行RNA测序以测量基因表达变化，来表征BTK抑制剂依鲁替尼和PI3K抑制剂AS-605240作为单一药物和联合药物在犬类临床前弥漫性大B细胞淋巴瘤（DLBCL）模型CLBL-1中的作用。
我们发现相对于对照组，在所有治疗类型和时间点上共有2336个差异表达基因（DEG）。差异表达基因数量最多的是依鲁替尼和AS-605240联合使用组。这些基因参与适应性免疫反应、白三烯D4代谢以及与GTP和GTPase介导的信号转导调节相关的术语。加权基因共表达网络分析（WGCNA）检测到九个基因模块，其中五个与治疗反应相关。这些模块中18%的基因也存在差异表达。值得注意的是，我们观察到一个模块仅与联合治疗相关，其基因成员与细胞代谢、稳态信号传导以及蛋白质合成和调节有关。
聚焦于与治疗反应相关的模块中高度连接的基因，这些基因在确定的主要靶向途径中发挥作用，且在不同治疗中有较大倍数变化，可作为协同治疗效果的潜在主要候选基因。
