title: Visual genome-wide RNAi screening to identify human host factors required for Trypanosoma cruzi infection.
abstract: The protozoan parasite Trypanosoma cruzi is the etiologic agent of Chagas disease, a neglected tropical infection that affects millions of people in the Americas. Current chemotherapy relies on only two drugs that have limited efficacy and considerable side effects. Therefore, the development of new and more effective drugs is of paramount importance. Although some host cellular factors that play a role in T. cruzi infection have been uncovered, the molecular requirements for intracellular parasite growth and persistence are still not well understood. To further study these host-parasite interactions and identify human host factors required for T. cruzi infection, we performed a genome-wide RNAi screen using cellular microarrays of a printed siRNA library that spanned the whole human genome. The screening was reproduced 6 times and a customized algorithm was used to select as hits those genes whose silencing visually impaired parasite infection. The 162 strongest hits were subjected to a secondary screening and subsequently validated in two different cell lines. Among the fourteen hits confirmed, we recognized some cellular membrane proteins that might function as cell receptors for parasite entry and others that may be related to calcium release triggered by parasites during cell invasion. In addition, two of the hits are related to the TGF-beta signaling pathway, whose inhibition is already known to diminish levels of T. cruzi infection. This study represents a significant step toward unveiling the key molecular requirements for host cell invasion and revealing new potential targets for antiparasitic therapy.
authors.affiliation: Image Mining Group, Institut Pasteur Korea, Seongnam-si, Gyeonggi-do, South Korea.
authors.forename: Auguste
authors.initials: A
authors.lastname: Genovesio
authors.forename: Miriam A
authors.initials: MA
authors.lastname: Giardini
authors.forename: Yong-Jun
authors.initials: YJ
authors.lastname: Kwon
authors.forename: Fernando
authors.initials: F
authors.lastname: de Macedo Dossin
authors.forename: Seo Yeon
authors.initials: SY
authors.lastname: Choi
authors.forename: Nam Youl
authors.initials: NY
authors.lastname: Kim
authors.forename: Hi Chul
authors.initials: HC
authors.lastname: Kim
authors.forename: Sung Yong
authors.initials: SY
authors.lastname: Jung
authors.forename: Sergio
authors.initials: S
authors.lastname: Schenkman
authors.forename: Igor C
authors.initials: IC
authors.lastname: Almeida
authors.forename: Neil
authors.initials: N
authors.lastname: Emans
authors.forename: Lucio H
authors.initials: LH
authors.lastname: Freitas-Junior
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000378:metabolism*; D002478:Cells, Cultured; D014355:Chagas Disease / Q000235:genetics* / Q000469:parasitology; D020869:Gene Expression Profiling; D015894:Genome, Human; D000882:Haplorhini; D057166:High-Throughput Screening Assays; D006790:Host-Parasite Interactions / Q000502:physiology*; D006801:Humans; D007668:Kidney / Q000166:cytology / Q000378:metabolism; D020411:Oligonucleotide Array Sequence Analysis; D034622:RNA Interference; D012333:RNA, Messenger / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016212:Transforming Growth Factor beta / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism*; D014349:Trypanosoma cruzi / Q000235:genetics
chemical_list: D015415:Biomarkers; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D016212:Transforming Growth Factor beta