Basak Swarnendu, Dutta Shovan, Khanal Supreet, Neelakanta Girish, Sultana Hameeda
Department of Biomedical and Diagnostic Sciences, College of Veterinary Medicine, University of Tennessee, Knoxville, TN, 37996, USA.
The Center for Immunotherapy and Precision Immuno-Oncology (CITI), Lerner Research Institute, Cleveland Clinic, Cleveland, OH, USA.
Sci Rep. 2025 May 30;15(1):19016. doi: 10.1038/s41598-025-02640-5.
Suppression of human megakaryocytes by dengue virus (DENV) infection significantly reduces the platelet count that eventually leads to thrombocytopenia, severe dengue and death. To understand DENV interactions with megakaryocytes, we investigated the cell cycle in leukemic human megakaryocytic in vitro cell line (MEG-01 cells). Megakaryocytes are known for complex endomitotic cell cycle leading to their polyploidy state. Our study shows that DENV uses these polyploid cells for its replication. Understanding the modulation of DENV-mediated cell cycle regulation in megakaryocytes is therefore highly important. We show that DENV2 (serotype 2) infection significantly modulates cell cycle signaling. Our protein profile microarray data showed significant upregulation of several cell cycle regulatory proteins including CDK4, CDK1, Cyclin B1 and others or downregulation of Chk1, GSK3-beta, CUL-3, and E2F-3. Quantitative real-time PCR and immunoblotting analyses further confirmed the upregulation of CDK4, CDK1, and Cyclin B1 upon DENV2 infection. Gene silencing of CDK4, CDK1 and Cyclin B1 showed significant reduction in DENV2 loads. Immunoprecipitation analysis further revealed an enhanced interaction between Cyclin B1 and CDK1 upon DENV2 infection that perhaps suggest the substantial changes noted in cell cycle regulation. Overall, our study suggests that DENV2 modulates cell cycle signaling in megakaryocytes and interferes with the critical regulatory proteins that may eventually lead to changes in endomitosis process. In conclusion, we report an important molecular insight regarding DENV2-mediated cell cycle modulation in human megakaryocytes.
登革病毒（DENV）感染对人巨核细胞的抑制作用会显著降低血小板计数，最终导致血小板减少、严重登革热甚至死亡。为了解DENV与巨核细胞的相互作用，我们在体外对白血病人类巨核细胞系（MEG - 01细胞）的细胞周期进行了研究。巨核细胞以其复杂的核内有丝分裂细胞周期导致多倍体状态而闻名。我们的研究表明，DENV利用这些多倍体细胞进行复制。因此，了解DENV介导的巨核细胞细胞周期调控的调节机制非常重要。我们发现，DENV2（血清型2）感染会显著调节细胞周期信号。我们的蛋白质谱微阵列数据显示，包括CDK4、CDK1、细胞周期蛋白B1等在内的几种细胞周期调节蛋白显著上调，而Chk1、GSK3 - β、CUL - 3和E2F - 3则下调。定量实时PCR和免疫印迹分析进一步证实了DENV2感染后CDK4、CDK1和细胞周期蛋白B1的上调。CDK4、CDK1和细胞周期蛋白B1的基因沉默显示DENV2载量显著降低。免疫沉淀分析进一步揭示了DENV2感染后细胞周期蛋白B1与CDK1之间的相互作用增强，这可能表明细胞周期调控中出现了实质性变化。总体而言，我们的研究表明，DENV2调节巨核细胞的细胞周期信号，并干扰关键调节蛋白，这最终可能导致核内有丝分裂过程发生变化。总之，我们报告了关于DENV2介导的人类巨核细胞细胞周期调节的重要分子见解。
