Entamoeba histolytica, a protozoan parasite, is an important human pathogen. Diseases caused by E. histolytica include dysentery and liver abscesses, and this organism is a leading parasitic cause of death on a global scale. The life cycle of Entamoeba involves stage inter-conversion between trophozoites and cysts. Importantly, the conversion to cysts allows the pathogen to disseminate to new hosts while development to trophozoites allows the organism to cause invasive disease in the host. Thus, stage interconversion is essential for disease transmission and pathogenesis. However, despite being central to amebic biology, stage conversion is poorly understood. Many factors have contributed to the paucity of data including the inability to reproduce the developmental cycle in E. histolytica. Instead, the reptilian ameba E. invadens, which has the same disease pathogenesis as E. histolytica, has been used as a model system to study Entamoeba development. In E. invadens, both encystation and excystation can be recapitulated with high efficiency in vitro. However, until recently a poorly annotated genome and lack of methods for genetic manipulation have prevented full exploitation of this system. The recent re-sequencing of the E. invadens genome, publication of the transcriptome of stage conversion, and development of methods for constitutive gene expression open up new avenues for discovery.
We aim to build upon these recent successes by establishing new tools for the study of E. invadens including (i) regulated gene expression, (ii) gene silencing, and (iii) development of negative selection and single cell markers. These methods will make possible new experimental strategies for studying Entamoeba stage conversion, and greatly aid the entire Entamoeba community. Dissecting the developmental cascade is crucial to understanding the biology of this important human pathogen.
Entamoeba histolytica is an important pathogen, which impacts human health on a global scale. Conversion between the trophozoite and cyst stage is central to parasite biology and responsible to disease dissemination and pathogenesis. Our long term goals are to understand the molecular mechanisms controlling stage conversion. Entamoeba development is best studied in Entamoeba invadens a reptilian ameba in which the developmental cascade is well established. However, the lacks of genetic tools in this system hinder progress. Thus, we aim to develop tools for regulated expression in E. invadens so that the molecular basis of the developmental cascade can be fully studied.
| Suresh, Susmitha; Ehrenkaufer, Gretchen; Zhang, Hanbang et al. (2016) Development of RNA Interference Trigger-Mediated Gene Silencing in Entamoeba invadens. Infect Immun 84:964-975 |
| Liu, Yuk-Chien; Singh, Upinder (2014) Destabilization domain approach adapted for regulated protein expression in the protozoan parasite Entamoeba histolytica. Int J Parasitol 44:729-35 |
| Manna, Dipak; Ehrenkaufer, Gretchen M; Singh, Upinder (2014) Regulation of gene expression in the protozoan parasite Entamoeba invadens: identification of core promoter elements and promoters with stage-specific expression patterns. Int J Parasitol 44:837-45 |
