Laboratorio de Biología Celular y Molecular; Instituto de Histología y Embriología (IHEM); Universidad Nacional de Cuyo; CONICET; Mendoza, Argentina.
Autophagy. 2013 Jul;9(7):1080-93. doi: 10.4161/auto.24709. Epub 2013 May 6.
Autophagy is a cell process that in normal conditions serves to recycle cytoplasmic components and aged or damaged organelles. The autophagic pathway has been implicated in many physiological and pathological situations, even during the course of infection by intracellular pathogens. Many compounds are currently used to positively or negatively modulate the autophagic response. Recently it was demonstrated that the polyamine spermidine is a physiological inducer of autophagy in eukaryotic cells. We have previously shown that the etiological agent of Chagas disease, the protozoan parasite Trypanosoma cruzi, interacts with autophagic compartments during host cell invasion and that preactivation of autophagy significantly increases host cell colonization by this parasite. In the present report we have analyzed the effect of polyamine depletion on the autophagic response of the host cell and on T. cruzi infectivity. Our data showed that depleting intracellular polyamines by inhibiting the biosynthetic enzyme ornithine decarboxylase with difluoromethylornithine (DFMO) suppressed the induction of autophagy in response to starvation or rapamycin treatment in two cell lines. This effect was associated with a decrease in the levels of LC3 and ATG5, two proteins required for autophagosome formation. As a consequence of inhibiting host cell autophagy, DFMO impaired T. cruzi colonization, indicating that polyamines and autophagy facilitate parasite infection. Thus, our results point to DFMO as a novel autophagy inhibitor. While other autophagy inhibitors such as wortmannin and 3-methyladenine are nonspecific and potentially toxic, DFMO is an FDA-approved drug that may have value in limiting autophagy and the spread of the infection in Chagas disease and possibly other pathological settings.
自噬是一种细胞过程,在正常情况下,它有助于回收细胞质成分和老化或受损的细胞器。自噬途径与许多生理和病理情况有关,甚至在细胞内病原体感染过程中也是如此。目前有许多化合物被用于正向或负向调节自噬反应。最近有研究表明,多胺亚精胺是真核细胞自噬的一种生理性诱导剂。我们之前曾表明,恰加斯病的病原体,原生动物寄生虫克氏锥虫,在宿主细胞入侵过程中与自噬隔室相互作用,并且自噬的预先激活显著增加了寄生虫对宿主细胞的定殖。在本报告中,我们分析了多胺耗竭对宿主细胞自噬反应和克氏锥虫感染性的影响。我们的数据表明,通过抑制生物合成酶鸟氨酸脱羧酶用二氟甲基鸟氨酸(DFMO)来消耗细胞内多胺,会抑制两种细胞系中因饥饿或雷帕霉素处理而诱导的自噬。这种效应与 LC3 和 ATG5 两种形成自噬体所需的蛋白质水平下降有关。由于抑制宿主细胞自噬,DFMO 损害了克氏锥虫的定殖,表明多胺和自噬促进了寄生虫的感染。因此,我们的结果表明 DFMO 是一种新型的自噬抑制剂。虽然其他自噬抑制剂,如渥曼青霉素和 3-甲基腺嘌呤是非特异性的,并且具有潜在的毒性,DFMO 是一种 FDA 批准的药物,可能在限制自噬和恰加斯病及其他可能的病理环境中的感染传播方面具有价值。