TY - JOUR
T1 - Visual genome-wide RNAi screening to identify human host factors required for Trypanosoma cruzi infection
AU - Genovesio, Auguste
AU - Giardini, Miriam A.
AU - Kwon, Yong Jun
AU - de Macedo Dossin, Fernando
AU - Choi, Seo Yeon
AU - Kim, Nam Youl
AU - Kim, Hi Chul
AU - Jung, Sung Yong
AU - Schenkman, Sergio
AU - Almeida, Igor C.
AU - Emans, Neil
AU - Freitas-Junior, Lucio H.
PY - 2011
Y1 - 2011
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=79956326721&partnerID=8YFLogxK
UR - https://pubmed.ncbi.nlm.nih.gov/21625474
U2 - 10.1371/journal.pone.0019733
DO - 10.1371/journal.pone.0019733
M3 - Article
C2 - 21625474
AN - SCOPUS:79956326721
SN - 1932-6203
VL - 6
JO - PLoS ONE
JF - PLoS ONE
IS - 5
M1 - e19733
ER -