克氏锥虫是一种具有极大医学意义的原生动物,因为它是恰加斯病的病原体,拉丁美洲的一种地方性疾病。这种寄生虫经历了表观遗传事件,比如磷酸化,甲基化和乙酰化,在包括复制在内的几个细胞过程中发挥作用,转录和基因表达。组蛋白脱乙酰酶(HDAC)参与细胞质蛋白的染色质压缩和翻译后修饰,如微管蛋白。TubastatinA(TST)是一种特定的HDAC6抑制剂,可影响细胞生长并促进癌细胞和寄生虫的结构修饰。在本研究中,我们证明,在TST治疗72小时后,Cruziepimastigote细胞的增殖和活力降低。通过不同的显微镜方法获得的结果表明,这种抑制剂会损害细胞骨架微管的聚合动力学,生成原生动物,表现出非典型的形态和细胞模式,包括多核寄生虫。此外,处理过的原生动物的微管被更强烈的乙酰化,特别是在细胞体的前部。细胞周期分析表明,G2/M期锥虫数量增加。一起,我们的结果表明,TST应该作为一种工具来研究锥虫细胞生物学,包括微管细胞骨架动力学,作为抗寄生虫药.
Trypanosoma cruzi is a protozoan of great medical interest since it is the causative agent of Chagas disease, an endemic condition in Latin America. This parasite undergoes epigenetic events, such as phosphorylation, methylation and acetylation, which play a role in several cellular processes including replication, transcription and gene expression. Histone deacetylases (HDAC) are involved in chromatin compaction and post-translational modifications of cytoplasmic proteins, such as tubulin. Tubastatin A (TST) is a specific HDAC6 inhibitor that affects cell growth and promotes structural modifications in cancer cells and parasites. In the present
study, we demonstrated that T. cruzi epimastigote cell proliferation and viability are reduced after 72 h of TST treatment. The results obtained through different microscopy methodologies suggest that this inhibitor impairs the polymerization dynamics of cytoskeleton microtubules, generating protozoa displaying atypical morphology and cellular patterns that include polynucleated parasites. Furthermore, the microtubules of treated protozoa were more intensely acetylated, especially at the anterior portion of the cell body. A cell cycle analysis demonstrated an increase in the number of trypanosomatids in the G2/M phase. Together, our results suggest that TST should be explored as a tool to
study trypanosomatid cell biology, including microtubule cytoskeleton dynamics, and as an antiparasitic drug.